Ebook Advances in Food and Nutrition Research PDF Full Chapter PDF

Advances in Food and Nutrition
Research - eBook PDF

Visit to download the full and correct content document:
https://ebooksecure.com/download/advances-in-food-and-nutrition-research-ebook-p
df/
ADVISORY BOARDS
David Rodríguez-Lázaro
Loong-Tak Lim
Michael Eskin
Isabel Ferreira
Crispulo Gallegos
Se-Kwon Kim
Keizo Arihara
SERIES EDITORS
GEORGE F. STEWART (1948–1982)
EMIL M. MRAK (1948–1987)
C. O. CHICHESTER (1959–1988)
BERNARD S. SCHWEIGERT (1984–1988)
JOHN E. KINSELLA (1989–1993)
STEVE L. TAYLOR (1995–2011)
JEYAKUMAR HENRY (2011–2016)
FIDEL TOLDRÁ (2016– )
Academic Press is an imprint of Elsevier
50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States
525 B Street, Suite 1650, San Diego, CA 92101, United States
The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom
125 London Wall, London, EC2Y 5AS, United Kingdom
First edition 2022
Copyright © 2022 Elsevier Inc. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or by any means, electronic
or mechanical, including photocopying, recording, or any information storage and retrieval system,
without permission in writing from the publisher. Details on how to seek permission, further
information about the Publisher’s permissions policies and our arrangements with organizations such
as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website:
www.elsevier.com/permissions.
This book and the individual contributions contained in it are protected under copyright by the
Publisher (other than as may be noted herein).
Notices
Knowledge and best practice in this field are constantly changing. As new research and experience
broaden our understanding, changes in research methods, professional practices, or medical
treatment may become necessary.
Practitioners and researchers must always rely on their own experience and knowledge in evaluating
and using any information, methods, compounds, or experiments described herein. In using such
information or methods they should be mindful of their own safety and the safety of others, including
parties for whom they have a professional responsibility.
To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume
any liability for any injury and/or damage to persons or property as a matter of products liability,
negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas
contained in the material herein.
ISBN: 978-0-323-99082-0
ISSN: 1043-4526
For information on all Academic Press publications

visit our website at https://www.elsevier.com/books-and-journals
Publisher: Zoe Kruze

Acquisitions Editor: Sam Mahfoudh
Developmental Editor: Jhon Michael Peñano
Production Project Manager:
Vijayaraj Purushothaman
Cover Designer: Matthew Limbert
Typeset by STRAIVE, India
Contributors
Alexios Alexopoulos
Laboratory of Agronomy, Department of Agriculture, University of the Peloponnese,
Kalamata, Messinia, Greece
Mikel Añibarro-Ortega
Centro de Investigação de Montanha (CIMO), Instituto Politecnico de Bragança, Bragança,
Portugal
Lillian Barros
Portugal
Edmundo Brito-de la Fuente
Product & Process Engineering Centre, Fresenius Kabi Deutschland GmbH, Bad Homburg,
Germany
Adriano Gomes da Cruz
Department of Food, Federal Institute of Education, Science and Technology of Rio de
Janeiro (IFRJ), Rio de Janeiro, RJ, Brazil
Isabel Diañez
Chemical Process and Product Technology Research Centre (Pro2TecS), Departamento de
Ingenierı́a Quı́mica, Universidad de Huelva, Huelva, Spain
N.A. Michael Eskin
Department of Food & Human Nutritional Sciences, University of Manitoba, Winnipeg,
MB, Canada
Isabel C.F.R. Ferreira
Portugal
Jose M. Franco
Crı́spulo Gallegos
Product & Process Engineering Centre, Fresenius Kabi Deutschland GmbH, Bad Homburg,
Germany
Loong-Tak Lim
Department of Food Science, University of Guelph, Guelph, ON, Canada
Bruna Marchesan Maran
Department of Chemical Engineering and Food Engineering, Federal University of Santa
Catarina, Technology Center, Florianópolis, Santa Catarina, Brazil
Emanueli Marchesan Maran
Department of Chemical Engineering and Food Engineering, Federal University of Santa
Catarina, Technology Center, Florianópolis, Santa Catarina, Brazil
ix
x Contributors
Inmaculada Martı́nez
Leticia Mora
Instituto de Agroquı́mica y Tecnologı́a de Alimentos (CSIC), Paterna, Spain
Ruchira Nandasiri
Department of Food & Human Nutritional Sciences, University of Manitoba; Richardson
Centre for Functional Foods & Nutraceuticals, Winnipeg, MB, Canada
Spyridon A. Petropoulos
Department of Agriculture, Crop Production and Rural Environment, University of
Thessaly, Volos, Greece
Milena Dutra Pierezan
Department of Food Science and Technology, Agricultural Sciences Center, Federal
University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
Tatiana Colombo Pimentel
Federal Institute of Paraná, Paranavaı́, Parana, Brazil
Jose Pinela
Portugal
David Rodrı́guez-Lázaro
Microbiology Division, Faculty of Sciences; Research Centre for Emerging Pathogens and
Global Health, University of Burgos, Burgos, Spain
Rachel Siqueira de Queiroz Simões
Institute of Technology in Immunobiologicals, Bio-Manguinhos, Oswaldo Cruz
Foundation, Fiocruz, Manguinhos, Rio de Janeiro, Brazil; Microbiology Division, Faculty of
Sciences; Research Centre for Emerging Pathogens and Global Health, University of Burgos,
Burgos, Spain
Fidel Toldrá
Silvani Verruck
Department of Food Science and Technology, Agricultural Sciences Center, Federal
University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
Amr Zaitoon
Department of Food Science, University of Guelph, Guelph, ON, Canada
Hongfei Zhang
Department of Food Science and Technology, National University of Singapore, Singapore,
Singapore
Weibiao Zhou
Department of Food Science and Technology, National University of Singapore, Singapore,
Singapore
Preface
The series Advances in Food and Nutrition Research has reached Volume 100,
and this represents a significant milestone after more than 70 years of the
series’ history. This series was initially named Advances in Food Research,
and the first volume was published in 1948. It had 459 pages distributed
across 10 chapters and was edited by Emil M. Mrak (editor until 1987)
and George F. Stewart (editor till 1982). The link to this volume is
https://www.sciencedirect.com/bookseries/advances-in-food-research/
vol/1/suppl/C. A new editor, C.O. Chichester was appointed in 1959,
who remained until 1988. The name of the series was changed to Advances
in Food and Nutrition Research in 1989 with Volume 33 and John E. Kinsella
who was the editor until 1993. At this point, the series was not published
on a yearly basis; however, in 2001, it started with one or two volumes being
published every year. In 1995, Steve L. Taylor was the editor, who remained
until 2011, followed by Jeyakumar Henry from 2011 to 2016. Then, Fidel
Toldrá took over as the editor in 2016 with Volume 76. Due to its success,
the series moved up to three volumes a year in 2009, then four volumes a year
in 2018, and currently approved to reach five volumes a year in 2023 and suc-
cessive years. Success continues as ScienceDirect usage experienced
a noticeable increase of 68% in the period 2016–2020. The number of cita-
tions is also increasing, with a CiteScore of 4.7 in 2019 that increased up to 7.0
in 2020, for this series in Q1, ranking 32 out of 310 publications in Food
Science with 89th percentile based on CiteScore data in Scopus.
This volume (Volume 100) contains eight chapters written by an inter-
national board of authors—some of them were previous guest editors of
serial thematic volumes including this series’ editor—and reports the latest
developments in relevant and interesting topics such as the use of pepti-
domics tools and their applications in bioactive peptides, the controlled
release of bioactives in food, the role of canolol as an antioxidant and anti-
cancer agent, the applications of Solanaceae in foods and nutraceuticals,
the latest developments in 3D printing of foods, the safe use of raw milk
in the processing of dairy foods, the new threats of enteric viruses, and,
finally, the use of low-energy X-ray for innovative nonthermal food
processing.
The first chapter deals with the use of peptidomics and the methodolo-
gies involved in the study of bioactive peptides, including their identification
xi
xii Preface
and quantitation, and in silico approaches. An analysis of posttranslational

modifications in peptides that can modify their physical and chemical prop-
erties is also presented, and the specific applications of peptidomics in
bioactive peptides are finally reported. The second chapter provides an over-
view of the basic controlled and triggered release concepts relevant to food
and active packaging applications. It presents approaches to encapsulate bio-
active compounds, their mode of release, the mass transport processes, and
the interactive carriers that are responsive to certain specific stimuli. The
third chapter focuses on the nature of canolol and its applications in food
and medicine. Canolol is a unique phenol formed during processing that
has antioxidant and anticancer properties. Specific extractability require-
ments from oilseed using different processing conditions and canolol deriv-
atives, including dimers and trimers, are discussed. The fourth chapter
discusses the nutritional value of the most important Solanaceae species
commonly used for their edible fruit, as well as those used in the develop-
ment of functional foods and nutraceuticals due to their bioactive constitu-
ents. The toxic and poisonous effects as well as the sustainable management
practices implemented to increase the added value of such crops are also dis-
cussed. The fifth chapter reviews the current advances in extrusion-based
3D food printing systems, presenting innovative 3D food printing systems
based on gelling and/or mixing. The future of extrusion-based 3D food
printing and its applications in the production of customized foods for specific
needs such as controlled nutritional or rheological properties are also dis-
cussed. The sixth chapter reviews the contaminants that may be present in
raw milk, especially conventional and emerging pathogens, antimicrobial-
resistant pathogens and commensal strains found in milk and dairy products,
and biological toxic substances formed by microorganisms like biogenic
amines and mycotoxins. The chapter also reports other chemical contami-
nants such as antibiotic residues, heavy metals, pesticides, polycyclic aromatic
hydrocarbons, melamine, dioxins, polychlorinated biphenyls, plasticizers, and
additives. The seventh chapter deals with the threads of enteric viruses due to
their high capacity for infection and preservation in food environments and
their difficulty for correct and sensitive detection. The chapter discusses the
current efforts toward prioritizing different aspects of their detection, epide-
miology, and control as well as the need to calibrate the current disinfection
procedures in the food industry. The eight and last chapter focuses on the use
of low-energy X-ray irradiation of foods for nonthermal microbial inactiva-
tion. This technology provides a higher microbial inactivation efficacy than
other irradiation techniques, and, therefore, low-energy X-ray irradiation
Preface xiii
constitutes an attractive alternative food preservation technique. Furthermore,

the current applications of low-energy X-ray, consumer acceptance, and
its limitations are discussed in this chapter.
In summary, this volume presents the combined efforts of 27 profes-
sionals with different backgrounds who are developing their research in a
variety of countries like Canada, Brazil, Singapore, Portugal, Greece,
Germany, and Spain. The editor thanks Jhon Michael Peñano (develop-
mental editor), Sam Mahfoudh (acquisition editor), the production staff,
and all the contributors for sharing their experiences and for making this
book possible.
FIDEL TOLDRÁ
Editor
CHAPTER ONE
Peptidomics as a useful tool

in the follow-up of food bioactive
peptides
Fidel Toldrá∗ and Leticia Mora
∗
Corresponding author: e-mail address: [email protected]
Contents
1. Introduction 2
2. Peptidomics in the characterization of peptides 3
2.1 Identification of peptides 7
2.2 Quantification of peptides 15
3. In silico approaches 19
4. Mechanisms of enzymatic hydrolysis of food proteins 22
5. Release of bioactive peptides in foods by endogenous peptidases 24
6. Release of bioactive peptides through food proteins hydrolysis 30
7. Conclusions 39
References 39
Abstract
There is an intense research activity on bioactive peptides derived from food proteins in
view of their health benefits for consumers. However, their identification is quite chal-
lenging as a consequence of their small size and low abundance in complex matrices
such as foods or hydrolyzates. Recent advances in peptidomics and bioinformatics are
getting improved sensitivity and accuracy and therefore such tools are contributing to
the development of sophisticated methodologies for the identification and quantifica-
tion of peptides. These developments are very useful for the follow-up of peptides
released through proteolysis either in the food itself through the action of endogenous
peptidases during processing stages like fermentation, drying or ripening, or from food
proteins hydrolyzed by commercial peptidases or microorganisms with proteolytic
activity. This chapter is presenting the latest advances in peptidomics and its use for
the identification and quantification of peptides, and as a useful tool for controlling
the proteolysis phenomena in foods and protein hydrolyzates.
Advances in Food and Nutrition Research, Volume 100 Copyright # 2022 Elsevier Inc. 1
ISSN 1043-4526 All rights reserved.
https://doi.org/10.1016/bs.afnr.2022.03.001
2 Fidel Toldrá and Leticia Mora
1. Introduction
It has been widely reported that food proteins constitute a good source
of bioactive peptides that remain inactive while forming part of the parent
protein, although they may turn active if released through enzymatic hydro-
lysis. Food bioactive peptides may exert health promoting beneficial effects
such as antioxidant, antihypertensive, antiinflammatory, hypoglycemic,
hypocholesterolemic, antimicrobial and antitumor activities (Toldrá,
Reig, Aristoy, & Mora, 2018). Such bioactivity information is available
in open access databases that report data about chemical and structural char-
acteristics of bioactive peptides and protein of origin, its IC50, and references
(Minkiewicz, Iwaniak, & Darewicz, 2019). The beneficial effects of bioac-
tive peptides naturally generated in foods, preventing infection and diseases
is of great interest in an aging population. This makes such foods rich in bio-
active peptides an attractive option in daily diet. Furthermore, bioactive
peptides produced through enzymatic hydrolysis in large amounts make
them attractive as food supplements and as functional components to regu-
late health.
Peptidomics techniques are mainly focused on the study of peptides,
including their identification and quantitation. The target peptides may have
been generated in different ways specially when they come from food matri-
ces (see Fig. 1). So, bioactive peptides may be released during key stages in
FOOD
PEPTIDOMICS PEPTIDES
BIOLOGICAL
SYSTEM
PROTEOMICS Endogenous
PEPTIDOMICS
enzymes
PROTEINS Commercial
enzymes
Gastrointestinal
enzymes
Fig. 1 Scheme of peptides generation from food proteins and main disciplines for the
study of proteins and peptides.
Peptidomics as a useful tool 3
food processing like fermentation, drying, and ripening of meat and dairy
products, wine, sauces among other (Corr^ea et al., 2014; Mohanty,
Mohapatra, Misra, & Sahu, 2016; Mora et al., 2015). Bioactive peptides
may be also produced through controlled enzymatic hydrolysis, with com-
mercial peptidases or microorganisms, of proteins obtained from different
types of food waste and by-products resulting from slaughterhouses, fisher-
ies, whey, fruits peels, etc. (Mora, Reig, & Toldrá, 2014; Ryder, Bekhit,
McConnell, & Carne, 2016), and also hydrolyzates from egg, soybean
and peanut proteins, among other (De Oliveira et al., 2015; Ji, Sun,
Zhao, Xiong, & Sun, 2014). Commercial peptidases are adequate to be
used in food and can be obtained from different origins such as microorgan-
isms, vegetable sources or animal organs. Finally, gastrointestinal digestion
must be taken into account because pepsin in the gastric step and trypsin,
chymotrypsin and pancreatic enzymes in the intestinal step are also able
to release bioactive peptides from the ingested proteins or even inactivate
some peptides due to further hydrolysis (Capriotti et al., 2015; Pepe
et al., 2016). Furthermore, an intense proteolytic activity due to the action
of brush border intestinal epithelium proteases and blood stream enzymes
that complete protein digestion acting as exopeptidases and releasing
dipeptides and free amino acids.
The recent improvements in sensitivity and accuracy achieved in
peptidomics instrumentation and advances in bioinformatics tools are
allowing the development of more sophisticated methodologies for the
identification of peptides. The knowledge of structure and function of
bioactive peptides naturally generated in foods and in hydrolyzates to be
used in food supplements and nutraceuticals, is essential for the optimization
of processing and quality control.
This chapter is presenting the latest advances in peptidomics as a useful
tool for a better understanding and control of the proteolysis phenomena
and therefore as an adequate tool for following the generation of bioactive
peptides and its quantitation in foods and hydrolyzates.
2. Peptidomics in the characterization of peptides

Peptidomics techniques have been successfully used for a better
understanding of gastrointestinal digestion process, the contribution of
endogenous and microbial enzymes in different food processes such as
fermentation (Li & Wang, 2021; Yu, Yu, & Jin, 2021), curing (Gallego,
Processing
Color, texture, flavour,

Biomarkers Quality
taste peptides
Authentification, OGMs,
Safety allergens, pathogensor
toxins
Bioactivities: antioxidant, antihypertensive,

hypoglycemic, hypercholesterolemic, etc.
Bioactives
Structure-activity
Peptidomics prediction
Post-traductional Oxidation, deamidation,

modifications etc.
Gastrointestinal
Intestinal microbiota
digestion
Protein-peptide
interactions
Fig. 2 Major areas of peptidomics applications in foods.
Mora, & Toldrá, 2016), aging (Kominami, Hayashi, Tokihiro, & Ushio,
2021; Renzone, Novi, Scaloni, & Arena, 2021), the identification of bioac-
tive peptides, and the search of peptide markers in food systems (see
Fig. 2). A recent interest in peptidomics is growing in the area of food waste
and by-products valorization because such peptidomics tools contribute to
a better understanding of the proteolysis phenomena and therefore a
better control and steering of the processes (Martini, Solieri, Cattivelli,
Pizzamiglio, & Tagliazucchi, 2021; Martini, Solieri, & Tagliazucchi, 2021).
Peptidomics, in conjunction with other disciplines like proteomics,
genomics, transcriptomics, and metabolomics, contributes to a better
understanding of the food systems. However, there are some difficulties that
make the study of the proteome very complicated such as the low-
abundance of peptides, their heterogeneity and variety in size, charges,
characteristics, and the complexity of biological matrices when they are
distributed. For this reason, a high amount of extraction, separation, isola-
tion, identification, and quantitation techniques have been developed and
used in the last years.
The description of main steps used to simplify, reduce complexity and
eliminate potential interferences in food samples, before the identification
Pretreatment
Pulsed Electric Fields/High
Ultrasounds/Microwaves Acid/base
Hydrostatic Pressure
Extraction
Precipitation/
Solid phase cartridges Acid/water/organic Centrifugation deproteinization
Separation/fractionation
Size-exclusion
Electrophoresis Liquid chromatography
chromatography
Purification
Fractions collection Ultrafiltration
Fig. 3 Description of main steps used to simplify, reduce complexity, and eliminate
potential interferences in food samples before the identification and quantitation of
bioactive peptides.
and quantitation of bioactive peptides is shown in Fig. 3. The pretreatment

step is used to remove potential interferences, as well as to facilitate the
extraction and concentration of peptides. Current tendencies are the use
of green and cost-effective technologies such as ultrasounds, microwaves,
pulsed electric fields, or high hydrostatic pressure, although chemical
pretreatments have been also described to result very effective in the later
extraction of peptides or proteins (Franca-Oliveira, Fornari, & Hernández-
Ledesma, 2021). The extraction procedure of peptides is done using water
or organic solvents depending on the expected characteristics of the generated
peptides. Polar peptides are easily extracted using aqueous and slightly
acidic solvents whereas hydrophobic sequences are extracted with organic
phases. The extraction of peptides may also carry part of protein fraction,
especially when aqueous solvents are used, and therefore a later step of
deproteinization by precipitation and centrifugation is often needed. The next
step is the use of one or a combination of different separation techniques in
order to fractionate the peptide extract at the same time that concentrate
the biologically active fraction. The most used methodologies are Tricine
SDS-PAGE electrophoresis that allows the separation of oligopeptides and
long peptides up to 2500 Da (Sch€agger, 2006) although when the interest
is in the separation of smaller peptides, chromatography techniques are the
method of choice. Size-exclusion chromatography is based on the separation

of peptide fractions according to their molecular weight, and the resolution of
separation depends on particle size, pore size, flow rate, column length and
diameter, and sample volume. Generally, the highest possible resolution is
obtained using moderate flow rates, long, narrow columns, and small particle
size gels, using same eluent in sample and mobile phase. Molecular weight
standards are used in column calibration in order to know the molecular
weights of unknown peptides in the extract. Also high resolution liquid chro-
matography (HPLC) is a frequently used methodology in the separation of
peptides according to their amino acid composition and characteristics (charge,
hydrophobic interaction, and molecular size). In this sense, reversed-phase
followed by hydrophilic interaction chromatography, and ion-exchange chro-
matography are the most used methods for peptide mixtures separations.
Reversed-phase C18 stationary phases are the most popular, and the elution
occurs by increasing the hydrophobicity of the mobile phase. However, this
separation not always results adequate when highly polar and small peptides are
in the mixture, and hydrophilic interaction chromatography (HILIC) have
resulted in very good results to separate small polar compounds on polar
stationary phases. This method can be complementary to reversed-phase as
the order of elution of the peptides are opposite to that found in reversed-
phase. The use of ion-exchange chromatography in the analysis of peptides
is limited to those applications that permit salt elimination in later steps since
moderate concentrations of salts could inhibit the ionization using electrospray
ionization (ESI) prior mass spectrometry analysis. After the chromatographic
separation using HPLC, eluting fractions are collected for subsequent purifi-
cation steps such as ultrafiltration with different molecular cut-off membranes
or to be directly analyzed by mass spectrometry.
Mass spectrometry methodologies have been widely developed during
the last decade and, currently, they have become the best choice for the
identification and quantification of bioactive peptides (Panchaud, Affolter,
& Kussmann, 2012). In this context, the advances in mass spectrometry
applications and the progressive improvement in the resolution of the instru-
ments have increased the development of peptidomics applications in the
food field, especially regarding the identification and quantitation of bioac-
tive peptides. Also the latest developments in bioinformatics are necessary to
obtain the maximum benefit from the generated data, including the use of
in silico tools, structure–activity relationship models, chemometrics, protein
and peptide databases and search engines, and statistics to manage the
enormous amount of data generated.
2.1 Identification of peptides

Traditionally, proteomics has been used to identify and quantify proteins
from different types of food matrices, obtaining the characterization of
the protein profile of the sample. The most frequently used strategy has
been the separation of proteins using gel electrophoresis and the subsequent
digestion of the isolated proteins with proteases with cleavage sites included
in the databases such as trypsin, Arg-C or Asp-N. The obtained hydrolyzates
are analyzed by mass spectrometry using specialized tools to obtain a list of
peaks to be processed using protein databases. Main approaches for the iden-
tification of proteins are based on Peptide Mass Fingerprint (PMF), where
the experimental data are a list of peptide mass values from the digestion of a
protein by a specific enzyme as previously indicated, and on MS/MS ions
search, where the identification is based on raw MS/MS data from one
or more peptides. Despite these approaches are very useful for the identifi-
cation of the presence of certain proteins, the experimental design needs
to be adapted when the objective is to analyze bioactive peptides obtained
from either natural processes such as fermentation, curing, or gastrointes-
tinal digestion, or commercial hydrolysis such as the treatment of food
by-products.
Bioactive peptides can exert different type of bioactivities depending on
different factors such as their physico-chemical characteristics, amino
acids sequence, structural conformation, etc. In fact, bioactive peptides have
been described to be able to modify or modulate different physiological sys-
tems such as the cardiovascular, digestive, endocrine, immune, muscular,
nervous, renal, reproductive, respiratory, and skeletal systems, due to their
potential to exert antihypertensive, antidiabetic, anticholesterolemic, anti-
oxidant, immunomodulatory, among other activities. Fig. 4 shows the main
Others
Opioid ACE inhibitor
Celiac toxic
DPP IV
inhibitor
Antioxidant
Antimicrobial
Fig. 4 Diagram showing main percentages of the most reported bioactive peptides
available in BIOPEP database; accessed on Feb, 2022 (Minkiewicz et al., 2019).
bioactivities studied according to BIOPEP database (Minkiewicz et al.,

2019), showing ACE-inhibitory activity as the most reported biological
activity followed by antioxidant and antimicrobial activity.
These peptides are very different and can show different characteristics
such as charge state, hydrophobicity or length, including large, medium,
and small peptides. However, small bioactive peptides (from 2 to 10 amino
acids length) represent by far the largest category as indicated in Fig. 5
(obtained from BIOPEP bioactive peptides database), probably due to their
small size makes them more adequate to cross the intestinal barrier and
achieve intact the blood stream and target organs to develop their biological
activity. Thus, despite the numerous difficulties that involve the analysis of
small peptides, more than half of the peptides described in the literature
and included in the BIOPEP database (Minkiewicz et al., 2019) are smaller
than 6 amino acids length (see Fig. 5).
The strategy for the identification of bioactive peptides will be different
considering the different physical and chemical properties of target peptides.
In this sense, separation methods used in mass spectrometry instruments and
informatics analysis will depend on the characteristics and size of peptides,
being in the interface between proteomics and metabolomics approaches.
In this regards, data analysis results very challenging as there is an estimation
of false positives 100 times higher for peptidomics than for proteomics
because peptidomics relies on a single peptide identification and requires
very high quality spectra and frequent de novo identification.
2.1.1 Identification of di-, tri-, and tetrapeptides

Very small peptides are of particular interest to be used as bioactive peptides,
but the analytical strategy for their identification must be carefully designed
Fig. 5 Number of amino acids of the bioactive peptides available in BIOPEP database;
accessed on 17 Feb, 2022 (Minkiewicz et al., 2019).
considering that the analysis of these small peptides are between the analysis
of peptides and small molecules.
First steps of isolation and purification need to be planned in order to
avoid losses in part of the target peptides. In fact, whereas some of these
peptides contain hydrophobic amino acids and are properly retained in
C18 columns, others are mainly hydrophilic and need other types of station-
ary phases to be retained. Thus, liquid chromatographic separation of
peptides before their injection into the mass spectrometry analysis
should have taken into consideration the physico-chemical characteristics
of the expected peptides, the potential mixture of different small peptides
in the sample extract, and the choice of the most appropriate separation
techniques, for instance, a combination of different separation techniques
such as reversed phase and hydrophilic interaction chromatography.
Concentration steps are necessary to increase the abundance of these pep-
tides that frequently found at very low concentration, but this step requires
special attention because it may often lead to the loss of some of the
sequences in the washing/cleaning step.
Once retention and isolation is achieved, the analysis of peptides by
mass spectrometry is another very critical step. The selection of suitable mass
analyzer depends on the applications of interest. When analyzing samples
with complex matrices, a quadrupole would be useful to reduce the inter-
ferences from the matrices. For further quantitative analysis of trace compo-
nents in complex samples, QqQ or Q-Orbitrap would be a good choice due
to their high sensitivity and quantitative ability. Orbitrap is also used to
obtain the accurate mass of the targeted molecules, which reveals the
elemental compositions of the molecules (Chang et al., 2021). In the iden-
tification of di- and tripeptides, it is necessary to use high mass accuracy and
high resolution instruments such as Quadrupole Time-of-Flight (Q-ToF) or
Triple Quadrupole (QQQ) analyzers, although QTrap can realize MSn
analysis, which is beneficial to qualitative analysis. In this regards, the main
difficulty is the optimization of the collision energy necessary for the frag-
mentation of the di- and tripeptides. In longer peptides, it is possible to get
several ion fragments and differentiate the precursor, whereas in small
peptides the amounts and quality of fragments are lower.
In fact, in the identification of dipeptides by mass spectrometry it is nec-
essary to consider retention time, precursor mass, and ion fragments in order
to establish an adequate correlation between this information and the
sequence of the peptide. In this sense, most studies focused on the identifi-
cation of small di and tripeptides also cover the quantitation of the
molecules.
Dipeptide sequences can be repeated many times in protein sequences so

they are not unique. For this reason, dipeptides identification by matching
the obtained spectrum with protein databases such as UniProt and NCBInr
is not feasible. Furthermore, current search algorithms are often limited to
peptides containing 4 or more amino acids, as smaller peptides would result
in unacceptable scores because the search engines are not prepared for
this use (Tang, Li, Lin, & Li, 2014). Thus, de novo identification of the
sequences by highly-experienced personnel is frequently needed, which
can be difficult and time-consuming, mostly for untargeted analysis
(Mora et al., 2017). However, this has been the method of choice of many
researchers although a limited amount of spectra can be analyzed.
The main proteomic approach used for the identification and quantita-
tion of small peptides is the Multiple Reaction Monitoring (MRM) using
QQQ instruments. Quadrupole is often placed in front of other mass
analyzers to pre-select targeted ions. Interferences from matrix ions are
removed, significantly reducing chemical noises. In general, QqQ mass
spectrometer has the highest sensitivity. The QqQ is the most commonly
used mass analyzer for quantitative detection due to its higher accuracy.
MRM also presents certain difficulties, as it needs to be previously opti-
mized, and requires trained personnel and expensive instruments.
On the other hand, mass spectrometry approaches based on matrix-
assisted laser desorption/ionization (MALDI) mass spectrometry have the
advantage of convenience, speed, and accuracy, but also present good
resolution, robustness, and sensitivity. Relative and absolute quantification
can be performed using MALDI approaches based on mass spectrometry in
tandem (MS/MS) (Wang & Giese, 2017). However, this methodology is
not very popular for the identification of small peptides mainly due to the
potential interferences of low-molecular peptides with MALDI matrices
used for ionization although the high sensitivity of MALDI-MS and its
tolerance against interference by salts and other components allowed for
the detection of native peptides from complex biological mixtures with
reduced sample preparation efforts. Recently, a MALDI Time-of-Flight
(ToF) approach was developed to achieve the fast detection of dipeptides
in dry-cured ham peptide extracts with the advantage of using very low
amounts of sample, while reducing time and cost (Heres, Saldaña,
Toldrá, & Mora, 2021; Heres, Yokoyama, et al., 2021). The characteristic
matrix ions appearing between 200 and 300 m/z were eliminated by sub-
traction using bioinformatics tools, and an example of MALDI-ToF MS
spectra is shown in Fig. 6. Later, the identification of the dipeptides was
confirmed by QQQ (Fig. 7).
Standard Mix
35000
a.i.
Standard Mix
30000
25000
AH VF
DD
20000
AL
EV
15000
10000
5000
190 200 210 220 230 240 250 260 270 280
m/z
Fig. 6 MALDI-ToF MS spectra of peptide mixture (Mix). Peptide sequences are given as
amino acid one-letter code. Source: Reproduced from Alejandro Heres, Celia Saldaña, Fidel
Toldrá, Leticia Mora, (2021). Identification of dipeptides by MALDI-ToF mass spectrometry in
long-processing Spanish dry-cured ham, Food Chemistry: Molecular Sciences, 3, 100048,
with permission from Elsevier.
2.1.2 Pentapeptides or longer peptides identification

Bioactive peptides between 5 and 30 amino acids length can be generated
either naturally due to the action of endogenous/digestive enzymes or
due to the action of commercial enzymes. The extraction and purification
strategy will depend on the characteristics of the peptides of interest and it
would be optimized according to their physico-chemical profile although
the use of C18 reversed phase columns is the most popular separation
methodology used in this type of peptides (see an example in Fig. 8).
However, whereas peptides obtained from the hydrolysis with commer-
cial enzymes can be more abundant due to hydrolysis conditions and have
been optimized to obtain a high recovery in bioactive peptides, those pep-
tides naturally generated by endogenous or gastrointestinal enzymes will
be at very low concentrations within complex biological matrices, and
therefore the analytical methods used for their identification have to be very
sensitive and rely on a specific sample preparation strategy. Regarding the
identification by mass spectrometry in tandem, traditional proteomics pro-
cedures could be used since the size of the peptides is similar to those sizes
obtained in traditional proteomics protocols using trypsin but with a certain
Fig. 7 ESI-QQQ spectra of the dipeptides AH, AL, DD, EV and VF identified in 18 months
dry-cured ham extracts. Source: Reproduced from Alejandro Heres, Celia Saldaña, Fidel
Toldrá, Leticia Mora, (2021). Identification of dipeptides by MALDI-ToF mass spectrometry
in long-processing Spanish dry-cured ham, Food Chemistry: Molecular Sciences, 3, 100048,
with permission from Elsevier.
adjustment of the basic techniques in order to identify a larger number of

endogenous peptides. Best option for the identification of these peptides
is the use of tandem mass spectrometry instruments with high resolution.
Ionization sources used are MALDI or ESI that are usually coupled to tan-
dem mass spectrometers such as quadrupole-ion trap, commonly used to
Fig. 8 Reversed-phase chromatographic separation after in vitro gastrointestinal diges-

tion of an Alcalase hydrolysate of orange by-products for the production of bioactive
peptides after gastrointestinal digestion. The figure shows the fractions from 44 to
54 obtained after SEC separation. The fractions were automatically collected and
assayed for their DPPH scavenging activity (A), ferric reducing capacity (B), ACEI-
inhibitory activity (C) α-amylase inhibitory activity. Reproduced from Mazloomi, S.N.;
Mahoonak, A.S.; Mora, L.; Ghorbani, M.; Houshmand, G.; Toldrá, F. Pepsin Hydrolysis of
Orange By-Products for the Production of Bioactive Peptides with Gastrointestinal
Resistant Properties. Foods 2021, 10, 679.
conduct MSn fragmentation of the parent peptides, quadrupole/time-of-

flight (Q/ToF), or time-of-flight/ time-of-flight (ToF/ToF) (Panchaud
et al., 2012). In this regards, Fig. 9 shows the distribution of peptides iden-
tified in dry-cured ham before and after gastrointestinal digestion according
to their protein of origin. In this example, chromatographic and mass
A) B)
Fig. 9 Distribution of the peptides identified by nLC-MS/MS according to their origin proteins in (A) undigested and (B) digested dry-cured
ham samples simulating in vitro gastrointestinal digestion. The identification of peptides was performed by nanoliquid
chromatography-tandem mass spectrometry (nLC-MS/MS) using a Nano-LC Ultra 1D Plus system (Eksigent of AB Sciex, CA, USA) coupled
to the quadrupole/time-of-flight (Q/ToF) TripleTOF® 5600+ system (AB Sciex Instruments, MA, USA) with a nanoelectrospray ionization
source (nESI). Reproduced from Gallego, M., Mauri, L., Aristoy, MC., Toldrá, F., Mora, L. (2020). Antioxidant peptides profile in dry-cured ham
as affected by gastrointestinal digestion, Journal of Functional Foods, 69, 103956, with permission from Elsevier.
spectrometry analysis revealed changes in the peptide profiles and evidenced

the degradation of proteins, mainly titin, by the digestive enzymes (Gallego,
Mauri, Aristoy, Toldrá, & Mora, 2020).
The data analysis approach has to be adapted according to the origin of
the peptides. Current search engines such as Mascot has been designed to
work with spectra data corresponding to peptides of sizes longer than 4
amino acids. However, mass spectrometry instrument parameters, peak list
parameters, and database search parameters have to be carefully selected in
order to get the maximum amount of identifications with high score. The
cleavage sites of bioactive peptides are not included in any of the descriptions
included in the search engines so frequently the choose of no-enzyme is
the unique option. This fact increases the search times and decreases the
specificity in the identification but good results could be obtained if all
parameters are well adjusted. It is recommended a previous optimization
of the parameters using a controlled protein digestion such as BSA. In order
to validate automatically obtained results using a search engine like Mascot
(https://www.matrixscience.com/search_form_select.html), several tools
have been developed such as the repetition of the search using identical
search parameters, against a database in which the sequences have been
reversed or randomized. Then, it is not expected to get any true matches
from the “decoy” database so the number of matches that are found is an
excellent estimation of the number of false positives that are present in
the results from the real or “target” database (Chen, Zhang, Xing, &
Zhao, 2009; Elias, Haas, Faherty, & Gygi, 2005).
2.2 Quantification of peptides

There is a lack of information about the concentration of the bioactive pep-
tides generated during food processing, the amount of peptides bioavailable
in the bloodstream after gastrointestinal digestion, and even the necessary
concentrations to establish an adequate dose to be used as ingredients. In fact,
the quantification of bioactive peptides faces up to several difficulties due to
their small molecular size, that is between metabolomics and proteomics dis-
ciplines, and the complexity of food matrices (Fig. 10). Thus, the use of
advanced mass spectrometry techniques is critical to identify the sequence
of these small peptides and to develop fast, precise and sensitive metho-
dologies for their quantitation. In peptidomics, the study of peptides often
includes their relative quantification with labeled or label-free methods.
The relative quantification of peptides is focused on the assessment of
peptide amounts in two or more samples, and the results are given by the
comparison of these amounts between samples. These methodologies
Fig. 10 Proteomics vs peptidomics. Main differences are in the generation and identi-
fication of peptides. Different approaches are needed when objectives are the identi-
fication of protein biomarkers and the identification of protein-derived bioactive
peptides. Reproduced from Mora, L., Gallego, M., Reig, M., Toldrá, F. (2017). Challenges
in the quantitation of naturally generated bioactive peptides in processed meats, Trends
in Food Science & Technology, 69, Part B, 306–314, with permission from Elsevier.
provide a simple, reliable, versatile, and cost-effective quantification that can

be based on peak intensity measurements or spectral counting (Bantscheff,
Schirle, Sweetman, Rick, & Kuster, 2007; Zhu, Smith, & Huang, 2010).
These approaches result very interesting in the comparison of peptides
profiles between different hydrolyzates as well as to elucidate the main pro-
teins responsible for the generation of bioactive peptides. In this regards,
Fig. 11 shows the results of a PCA analysis of the peptides profile obtained
from dry-cured ham by-products cooked and digested, and this label-free
analysis allowed the identification and relative quantitation of those peptides
responsible for the antioxidant activity detected in samples after 1 h of
cooking, which were collagen peptides (Gallego, Mora, Hayes, Reig, &
Toldrá, 2017).
On the other hand, other approaches such as multiple reaction monitor-
ing (MRM) provide the most accurate quantitative values for bioactive pep-
tides, but they require complex experimental protocols. MRM is a highly
sensitive and selective method of directed MS that uses three quadrupoles
for the analysis of short sequences and low concentration peptides in com-
plex mixtures (Hu et al., 2011; Nakashima et al., 2011; Priyanto et al., 2015;
Rawendra et al., 2014). Main important challenge of this procedure is the
Fig. 11 (A) Principal Component Analysis (PCA) score plot to assess the variance among
all the peptides of digested samples H2O, 100 °C 20 min, and 100 °C 1 h in three repli-
cates (n ¼ 3). Discriminant component 1 (t[1]) and discriminant component 2 (t[2])
explained a 52.9% and 20% of variability in the dataset, respectively. (B) PCA loading
plot showing the proteins of origin of those peptides more responsible for main differ-
ences between the uncooked and cooked samples after digestion. Reproduced from
Gallego, Mora, Hayes, Reig, Toldrá, Effect of cooking and in vitro digestion on the antiox-
idant activity of dry-cured ham by-products, Food Research International, 97, 2017,
296–306, with permission from Elsevier.
Fig. 12 Graphical abstract including the whole procedure for the identification, quan-
titation and confirmation of Ala-Ala dipeptide as antihypertensive peptide. Reproduced
from Heres, Yokoyama, Gallego, Toldrá, Arihara, Mora, (2021) Antihypertensive potential
of sweet Ala-Ala dipeptide and its quantitation in dry-cured ham at different processing
conditions. Journal of Functional Foods, 87, 104818, with permission from Elsevier.
optimization of the MRM parameters of low abundant peptides or small

size peptides to get a final accurate and sensitive quantification without inter-
ferences from other peptides or signal suppression (Capriotti, Cavaliere,
Piovesana, Samperi, & Lagana, 2016). Fig. 12 includes main results obtained
after the quantification of the antihypertensive dipeptide AA at different

times of processing of dry-cured ham using a triple quadrupole instrument.
3. In silico approaches
An example of a classical empirical approach to identify and confirm
the presence of bioactive peptides is shown in Fig. 13. This includes the
hydrolysis step to obtain bioactive peptides followed by their extraction
and later chromatographic separation. Frequently, several chromatographic
techniques are used for a better isolation of the peptides of interest. In fact,
the previous step to mass spectrometry identification usually consists of
liquid chromatography separation of the already isolated peptides. After
identification by mass spectrometry, those peptide sequences apparently
responsible for the biological activities are synthesized and the bioactivity
is confirmed in vitro and in vivo.
Protein of origin Food matrix

Hydrolysis Extraction
Peptides
1st fractionation
Isolation of bioactive fractions (SEC, CE, LC, GF-IEF,...)
In vitro test
2nd fractionation
Purification of peptides of interest (HPLC)
In vitro test
Identification by MS/MS
In vitro test
Synthesis of peptides
Bioactivity test in vitro and in vivo

Confirmation
Fig. 13 Scheme of the traditional empirical procedure for the identification and confir-
mation of bioactive peptides from food matrices. SEC, size-exclusion chromatography;
CE, capillary electrophoresis; LC, liquid chromatography; IEF, isoelectric focusing; HPLC,
high-performance liquid chromatography; MS/MS, mass spectrometry in tandem.
Reproduced from Mora, Gallego, Toldrá, F. (2018) ACEI-Inhibitory Peptides Naturally
Generated in Meat and Meat Products and Their Health Relevance. Nutrients, 10, 1259.
Despite this procedure is very effective, it results very expensive and

time-consuming. For this reason, modern in silico strategies based on simu-
lation using bioinformatics tools in most of the steps and peptide databases
are of high interest nowadays (see Fig. 14).
There are different tools that comprise from the selection of the protein
by determining the occurrence frequency of bioactive sequences in the pro-
tein in order to get bioactive peptide sequences after hydrolysis such as
PATTINPROT from PBIL server (https://npsa-prabi.ibcp.fr/cgi-bin/) or
BIOPEP (https://biochemia.uwm.edu.pl/). In this sense, BIOPEP database
contains a tool that permits the scanning of a protein sequence searching for
one or several patterns previously established to show the potential of the
protein to be a source of bioactive peptides (A) and the calculation of
the potential biological activity of each protein (B). The frequency of bio-
active fragments occurrence in protein sequence (A) is calculated using the
formula:
a
A¼
N:
where a is the number of fragments with given activity in a protein sequence,

and N is the number of amino acid residues of protein.
Fig. 14 Main steps of in silico approaches and open access databases for the selection of
the protein, hydrolysis simulation and bioactivity prediction. Reproduced from Mora,
Gallego, Toldrá, F. (2018) ACEI-Inhibitory Peptides Naturally Generated in Meat and
Meat Products and Their Health Relevance. Nutrients, 10, 1259.
The potential biological activity of protein (B) [μM1] is calculated with

the formula:
i
Σki¼1 ECa 50i
B¼
N
where ai is the number of repetitions of i-th bioactive fragment in protein

sequence, EC50i is the concentration of i-th bioactive peptide corresponding
to its half-maximal activity [μM] or half-maximal inhibition (IC50) in case of
peptides with inhibitory activity, k is the number of different fragments
with given activity, and N is the number of amino acid residues.
On the other hand, there are different tools designed to obtain theoret-
ical fragments from protein sequences by using enzymes with known
cleavage specificities. The hydrolysis of the selected protein is simulated
to generate in silico peptide profiles using bioinformatics tools such as
BIOPEP, where it is possible to simulate the digestion with up to three dif-
ferent enzymes simultaneously; PeptideMass from ExPASy (Gasteiger et al.,
2005; Wilkins et al., 1997), very intuitive and extended; or PoPS (Boyd,
Garcia de la Banda, Pike, Whisstock, & Rudy, 2004), a tool that allows
the prediction of substrate cleavage using protease specificity models that
have to be uploaded by the user.
However, the prediction when using commercial enzymes such as
Alcalase, Protamex, Neutrase, etc. is more complicated because they are
broad-spectrum endopeptidases. In this cases, the hydrolysis pattern of these
enzyme needs to be characterized in the laboratory to determine the poten-
tial fragments that would be obtained after optimal conditions of hydrolysis.
The online tool EnzymePredictor (http://bioware.ucd.ie/enzpred/
Enzpred.php) evaluates the evidence for which enzymes are most likely
to have cleaved a sample containing peptides from hydrolyzed proteins,
which would result very useful to drive hydrolysis processes toward the gen-
eration of certain specific bioactive peptides (Dalkiran et al., 2018). Also the
tool Proteasix is a web-based, peptide-centric tool (http://proteasix.cs.man.
ac.uk/index.html) dedicated to proteolytic events involved in naturally
occurring peptide generation. Proteasix takes a peptide list from the user
and then automatically reconstructs the N- and C-terminal cleavage sites
and identifies the observed and predicted proteases involved in the proteol-
ysis of these cleavage sites in three main species: Homo sapiens, Mus musculus
and Rattus norvegicus. This tool will allow to determine the potential enzymes
that are responsible for the generation of certain cleavage sites and results
very useful in the characterization of commercial enzymes main actions.
Once the theoretical peptide sequences are known, they can be analyzed
to identify desirable amino acids at certain position or interesting character-
istics that make them potential candidates to exert bioactivity by using
the software PeptideRanker (http://distilldeep.ucd.ie/PeptideRanker/),
which gives a list of scores that identifies those peptides that may be more
likely to be bioactive. This predictive tool is based on such general shared
features of bioactive peptides across different functional classes and aids in
the improved design of existing bioactive peptides (Mooney, Haslam,
Pollastri, & Shields, 2012).
Also there are many tools that predict structure and potential biological
activities. In this sense, some authors have reported the optimization of
Quantitative Structure–Activity Relationship (QSAR) models for revealing
relationships between structural properties of chemical compounds and bio-
logical activities. SAR modeling is essential for drug discovery but it is also
being used in bioactive peptides characterization (Kwon, Bae, Jo, & Yoon,
2019). Finally, the prediction of the three-dimensional structure of peptides
from their amino acid sequence and post-transductional modifications is
very important in peptidomics because the structure of the peptide can affect
its functionality. Thus, on-line tools such as PEPstrMOD (http://osddlinux.
osdd.net/raghava/pepstrmod/) (Singh et al., 2015) or PEP-FOLD (http://
bioserv.rpbs.univ-paris-diderot.fr/services/PEP-FOLD/) for long peptides,
allow the de novo prediction of multiple peptide structures for linear and
cyclic peptides (Lamiable et al., 2016; Shen, Maupetit, Derreumaux, &
Tuffery, 2014; Thevenet et al., 2012).
4. Mechanisms of enzymatic hydrolysis of food proteins

Proteolysis phenomena occurs in proteins that are hydrolyzed during
food fermentation and/or ripening. Endo- and exo-peptidases are the main
enzymes responsible for proteolysis and can be present in the food as either
endogenous enzymes or as microbial enzymes from the microorganisms
responsible for fermentation.
Proteins are first broken down by endopeptidases releasing protein frag-
ments and polypeptides. When such polypeptides are further fragmented
into smaller peptides, they act as substrates for further hydrolytic action
by exo-peptidases that will generate smaller tri- and dipeptides and free
amino acids (Toldrá et al., 2018). In this sense, peptidomics constitutes a
basic tool for the identification and quantification of the released peptides
and therefore the follow-up of the resulting peptide profiles (Mora
et al., 2017).
A scheme of how peptidases act on proteins is shown in Fig. 15. In such
example, endo-peptidases act on an internal linkage Phe-Pro (Mora,
Gallego, & Toldrá, 2019). The exo-peptidases may act on either the amino
or carboxy terminal. If a tripeptide is released, then they are named
tripeptidylpeptidases (TPP) and if it is a dipeptide, dipeptidylpeptidases
(DPP). For instance, X-prolyl dipeptidyl peptidase (PepX) releases dipep-
tides X-proline from the amino terminal. Tripeptidases may hydrolyze a
tripeptide into a dipeptide and a single amino acid while dipeptidases hydro-
lyze dipeptides into its two single amino acids (Toldrá, Gallego, Reig,
Aristoy, & Mora, 2020a). However, the major release of free amino acids
is caused by aminopeptidases (i.e., Pep N, Pep A, Pep C, Pep P or others)
acting on the amino terminal or by carboxypeptidases (A or B) acting on the
carboxy terminal (Mora, Gallego, Aristoy, & Toldrá, 2015).
The generation of bioactive peptides can take place in the food itself as
a consequence of the action of endogenous enzymes and autochthon-
ous microorganisms, and also during gastrointestinal digestion due to brush
A D E C C P C C
Thr Val Lys Glu Asp Gln Val Phe Pro Met Asn Pro Pro Lys Phe Asp Lys Ile Glu Asp
TVKEDQVFPMNPPKFDKIED
PPKFDKIED
TVKEDQVFPMNPPKFDKIED
VKEDQVFPMNPPKFDKIED
EDQVFPMNPPKFDKIED
VKEDQVFPMNPPKFDKIED
VKEDQVFPMNPPKFDKIE
VKEDQVFPMNPPKFDKI
TVKEDQVFPMNPPKFD
TVKEDQVFPMNPPK
TVKEDQVFPMNPP
Fig. 15 Scheme of food protein hydrolysis and enzymes involved. The

amino acids sequence is a fragment belonging to myosin heavy chain.
Aminopeptidase (A), Dipeptidylpeptidase (D), Endopeptidase (E), Carboxypeptidase
(C) and Peptidylpeptidase (P). Reproduced from Toldrá, F., Gallego, M., Reig, M.,
Aristoy, M.-C., Mora, L. (2020a). Recent progress in enzymatic release of food-derived
peptides and assessment of bioactivity. Journal of Agricultural & Food Chemistry, 68,
12842–12855, with permission from ACS.
Proteins in foods Isolated food proteins
Endogenous peptidases Microbial peptidases Peptidases Microbial peptidases
Ripening Fermentation Reactor Fermenter
Small amounts of bioactive Large amounts of bioactive

peptides within the food peptides as extracts
Gastrointestinal digestion
Absorption in the organism

Resistance to serum peptidases
Bioactive peptides
Physiological functions
Fig. 16 Scheme of the generation of bioactive peptides from protein hydrolysis in foods
and/or the hydrolysis of isolated food proteins. Reproduced from Toldrá, F., Reig, M.,
Aristoy, M.C., Mora, L. (2018). Generation of bioactive peptides during food processing.
Food Chemistry, 267, 395–404, with permission from Elsevier.
border intestinal epithelium proteases and blood stream enzymes that

complete protein digestion acting as exopeptidases. It can also be carried
out on food proteins previously extracted from foods or food by-products
and hydrolyzed with commercial peptidases (see Fig. 16). Both cases are
described below.
5. Release of bioactive peptides in foods by endogenous

peptidases
Food proteins may be hydrolyzed by endogenous or microbial pepti-
dases during ripening and/or fermentation (see Fig. 16). The result of such
proteolysis is the generation of numerous peptides and some of them may be
bioactive. Of course, such peptides must keep the bioactivity after the gas-
trointestinal digestion and absorption through the intestinal membrane in
order to exert its physiological benefit in the body (Gallego, Grootaert,
et al., 2016).
The action of endogenous peptidases is typical in long ripened foods like
meat and dairy products. For instance, in the case of meat products there
are several muscle endopeptidases, such as calpains and cathepsins, able to
hydrolyze myofibrillar and sarcoplasmic proteins into large protein

fragments and polypeptides. Such polypeptides constitute the substrates
for further hydrolysis by muscle exopeptidases such as tri- and
dipeptidylpeptidases, amino and carboxypeptidases (Mora, Escudero,
Arihara, & Toldrá, 2015; Mora, Sentandreu, & Toldrá, 2011; Toldrá,
Aristoy, & Flores, 2000) although such enzymes may be partly inhibited
by salt added during processing (Toldrá, Cerveró, & Part, 1993).
DPP I and II that are active at pH 5.5–6.5, near the pH found in most
meat products, can release dipeptides such as AG, AQ, RG, NP, PA, IL, SG,
SQ, among other from the N-terminal. TPP I, also active at pH 5.5–6.5,
releases specific tripeptides like RGA, IIP, GNP, GAG, GPG from the
N-terminal. As a result of dipeptidylpeptidases action during the processing
of dry-cured ham, dipeptides PA, GA, VG, EE, ES, DA, and DG were
reported to be generated (see Table 1), most of them with ACE inhibitory
activity and released in amounts ranging from 2 to 45 μg/g in dry-cured
ham (Heres et al., 2022). Dipeptides have important advantages for
biodisponibility and therefore are able to reach target organs because they
have small size and good resistance to further peptidase hydrolysis during
gastrointestinal digestion, intestinal absorption and circulation in the blood-
stream (Pentzien & Meisel, 2008).
Meat products like dry-cured ham, that have long processing times
like 9–12 months or longer, present numerous bioactive peptides
(Gallego et al., 2019; Toldrá, Gallego, Reig, Aristoy, & Mora, 2020b).
So, several ACE inhibitory peptides like AAATP and TLKRVP were iso-
lated from Spanish ham (Escudero et al., 2013; Gallego, Mora, & Toldrá,
2018), while peptides LGL, GVVPL and SFVIT were isolated from
Parma ham (Dellafiora et al., 2015). Numerous antioxidant peptides have
been also reported like GKFNV, FLKMN, and LPGGGHGNL in Jinhua
ham (Zhu et al., 2016), DLEE in Xuanwei ham (Xing et al., 2016),
AEEEYPDL and SNAAC in Spanish ham (Gallego, Mora, Reig, &
Toldrá, 2018; Gallego, Mora, & Toldrá, 2018). It is also noteworthy
the DPP IV inhibitory activity of some peptides isolated from Spanish
ham such as AAAAG, ALGGA and LVSGM (Gallego et al., 2014). An
extract with 36 peptide sequences from Xuanwei dry-cured ham was
reported exerted anti-inflammatory effect in RAW264.7 macrophage
cells, suppressing the secretion of nitric oxide, interleukin-6 and tumor
necrosis factor-α in the lipopolysaccharide peptides GPAGPL and
GPPGAP were those exhibiting the highest anti-inflammatory activity
(Fu et al., 2021).
Table 1 Examples of peptides released in foods during processing through the action of endogenous enzymes and microorganisms.
Food Type/Fermentation Peptide sequences Potential activity References
Dry-cured Chinese Jinhua FLKMN, GKFNV, Antioxidant Zhu, Zhang, Zhou, and Xu
ham LPGGGHGDL (2016)
Chinese Xuanwei DLEE Antioxidant Xing et al. (2016)
GPAGPL, GPPGAP Antiinflammatory Fu et al. (2021)
Italian Parma GVVPL, LGL, SFVIT ACE inhibitory Dellafiora et al. (2015)
Spanish SNAAC, AEEEYPDL Antioxidant Gallego, Mora, Reig, and
Toldrá (2018) and Gallego,
Mora, and Toldrá (2018)
FNMPLTIRITPGSKA Antiinflammatory, Gallego, Mora, and Toldrá
ACE inhibitory (2019)
HCNKKYRSEM, Antimicrobial, Castellano et al. (2016), Gallego
MDPKYR antiinflammatory, et al. (2019)
antioxidant, ACE
inhibitory
TKYRVP, Antiinflammatory, Gallego et al. (2019)
TSNRYHSYPWG antioxidant, ACE
inhibitory
AAATP ACE inhibitory Escudero et al. (2013)
AAAAG, ALGGA, LVSGM DPP IV inhibitory Gallego, Aristoy, and Toldrá
(2014)
PA, GA, VG, ES, DA ACE inhibitory Heres, Saldaña, Toldrá, and
Mora (2022)
Mutton ham MWTD, APYMM, FWIIE Antioxidant Wang, Lu, Li, Wang, and Huang
(2020)
Fermented Koji (Aspergillus, 10%, 30 °C, 24w) QYP Antioxidant Ohata, Uchida, Zhou, and
meat sauce Arihara (2016)
Cheese Italian Stracchino AVPYPQ, EAMAPK Antioxidant Pepe et al. (2016)
Parma cheese VPP, IPP, LHLPLP, HLPLP, ACE inhibitory Solieri et al. (2020)
ENLLRF
HLPLP, LHLPLP ACE inhibitory Basiricò et al. (2015)
APFPE DPP IV inhibitory Martini, Solieri, Cattivelli, et al.
(2021) and Martini, Solieri, and
Tagliazucchi (2021)
VPP, IPP, RYLGY, RYLG, ACE inhibitory Basiricò et al. (2015)
AYFYPEL, AYFYPE,
LHLPLP, and HLPLP
Brazilian Canastra artisanal Minas RPKHPIKHQG, Antimicrobial Fialho et al. (2018)
RPKHPIKHQ
Hard cow milk cheese EIVPN Antioxidant Timon, Andres, Otte, and
Petron (2019)
DKIHPF Antioxidant Timon et al. (2019)
VAPFPQ Antioxidant Timon et al. (2019)
Brazilian Prato/Lactobacillus helveticus QEPVLGPVRGPFPIIV ACE inhibitory Baptista et al. (2018)
(10%, 40 °C, 18 h)
YQEPVLGPVRGPFP ACE inhibitory Baptista et al. (2018)
Yoghurt Chinese Feng Wei Suan Ru/ FVAPFPEVF, PPFLQPEVM Antidiabetic, ACE Jin et al. (2016)
Streptococcus thermophellolus inhibitory
+ Lactobacilus bulgaricus
QEPVLGPVRGPFPIIV ACE inhibitory Jin et al. (2016)
Continued
Table 1 Examples of peptides released in foods during processing through the action of endogenous enzymes and microorganisms.—cont’d
Food Type/Fermentation Peptide sequences Potential activity References
Probiotic yoghurt with pineapple SLPQNIPPLTQTPVVVPPF Antioxidant, anticancer Sah, Vasiljevic, McKechnie, and
peel/S. thermophilus + L. bulgaricus Donkor (2016)
+ L. acidophilus + L. casei + L. paracasei
YQEPVLGPVRGPFPIIV Antioxidant, anticancer Sah et al. (2016)
(1%, 42 °C, pH 4.5)
Fermented Lactobacillus, Saccharomyces IPP, VPP ACE inhibitory Fekete, Givens, and Lovegrove
milk (2015)
Antiinflammatory, Chakrabarti and Wu (2015)
adipogneic
Antidiabetic Chakrabarti, Jahandideh,
Davidge, and Wu (2018)
Kluyveromyces marxianus (6%, 32 °C, LRFF, VLSRYP ACE inhibitory Li, Sadiq, Liu, Chen, and He
pH 6.5, 48 h) (2015)
Kombucha culture (1%, 37 °C, 72 h) FVAPEPFVFGKEK, ACE inhibitory Elkhtab, El-Alfy, Shenana,
LVYPFPGPLH, Mohamed, and Yousef (2017)
VAPFPEVFGK
L. actobacillus casei (1%, 37 °C, 72 h) LVESPPELNTVQ, ACE inhibitory Elkhtab et al. (2017)
VLESPPELN,
WGYLAYGLD
Fermented Lactobacillus pentosus (28 °C, 43d) IPP, KP, LPP, VPP ACE inhibitory Fideler, Johanningsmeier,
cucumber Ekel€
of, and Muddiman, 2019)
pickles
Fermented fish Malaysian pekasam/Lactobacillus AIPPHPYP, IAEVFLITDPK Antioxidant Najafian and Babji (2018)
plantarum (27 °C, 15d)
Fermented Thai Kapi Ta Dam IF, SV ACE inhibitory Kleekayai et al. (2015)
shrimp pastes
Thai Kapi Ta Dam, Kapi Ta Deang WP Antioxidant Kleekayai et al. (2015)
The use of starter cultures is an extended practice today for most fer-
mented foods that contributes to improve their safety and quality. The
microorganisms used as starter cultures contain a complex enzyme system
that may include peptidases (Flores & Toldrá, 2011) and their action may
result in the generation of bioactive peptides with beneficial health effects
(Martı́nez-Villaluenga, Peñas, & Frı́as, 2017). Particular attention is given
to lactic acid bacteria (LAB), microorganisms that have a high proteolytic
activity due to their content in extra and intracellular peptidases, that are
typically used for food fermentation. So, PepX, tripeptidase, dipeptidase,
PepN, PepA, PepC and other aminopeptidases have been reported in
LAB (González, Sacristán, Arenas, Fresno, & Tornadijo, 2010, Sinz &
Schwab, 2012; Stressler, Eisele, Kranz, & Fischer, 2014; Stressler et al.,
2016). Depending on the type and balance of such peptidases, different
peptide patterns are obtained in fermented foods and this may give
different bioactivity (Martı́nez-Villaluenga et al., 2017). For instance, cell
free extracts of Lactobacillus paracasei subsp. paracasei demonstrated X-prolyl
dipeptidylpeptidase, aminopeptidase, dipeptidase and carboxypeptidase
activities while Leuconostoc mesenteroides subsp. mesenteroides exhibited
endopeptidase, aminopeptidase, dipeptidase and carboxypeptidase activities
(Macedo, Vieira, Poças, & Malcata, 2010). Therefore, large amounts of
bioactive peptides may be expected in fermented foods. For instance, the
use of Lactobacillus pentosus and Staphylococcus carnosus in dry-fermented sau-
sages containing sodium caseinate as ingredient contributed to large amounts
of peptides with ACE inhibitory activity (Mora, Gallego, Escudero, et al.,
2015). Both microorganisms, Lactobacillus pentosus and Staphylococcus carnosus
are used for fermenting milk because both are able to hydrolyze casein
through their extracellular proteinase and the generated oligopeptides are
hydrolyzed by intracellular peptidases into smaller peptides once transported
into the cell (Chaves-López et al., 2014). Two antioxidant peptides were
reported after the simulated gastrointestinal digestion of soft cheese (Pepe
et al., 2016). Lactobacillus helveticus and Lactobacillus acidophilus are able to
hydrolyze Ƙ-casein and release short peptides, some with ACE inhibitory
activity (Ali et al., 2019). Other microorganisms of interest, typically used
for food fermentation and able to hydrolyze proteins, are yeasts (Santos
et al., 2001). So, peptidases such as PepX, leucine aminopeptidase, and
DPP IV and V have been reported in Aspergillus oryzae
(Matsushita-Morita et al., 2011), while proteinases A and D, and prolyl
and arginyl aminopeptidases were reported in Debaryomices hansenii
(Santos et al., 2001).
The presence of dipeptides have been reported in fermented foods as a

result of microbial dipeptidyl peptidases (DPP) hydrolysis on the amino
terminal. So, this is the case of dipeptides AF, PL, KL, LG and KF released
in Feta cheese by Lactobacillus paracasei (Bintsis, Vafopoulou-Mastrojiannaki,
Litopoulou-Tzanetaki, & Robinson, 2003), proline-containing dipeptides
released in sourdoughs by Leuconostoc mesenteroides and Lactobacillus curvatus
(Zotta, Ricciardi, & Parente, 2007) and by Lactobacillus helveticus in casein
hydrolysates (Stressler, Eisele, & Fischer, 2013), RP and GF by
Leuconostoc mesenteroides subsp. mesenteroides, and GP by Lactobacillus paracasei
subsp. paracasei in Serra da Estrela cheese (Macedo et al., 2010).
The release of bioactive peptides during food processing has been
widely documented, especially in milk, cheese and dairy foods, meat and
meat products, wine, fish and seafood, eggs, and other foods as shown in
Table 1. A large number of short peptides (>100) were identified, as endog-
enously released, in European sea bass and most of them were bioactive
(Cerrato et al., 2021). ACE inhibitory peptides VPP and IPP were reported
to increase with the ripening time in semi-hard cheeses (Meyer, Butikofer,
Walther, Wechsler, & Sieber, 2009). Two hexapeptides with relevant
antioxidant activity were isolated and identified after the simulated gastro-
intestinal digestion of Stracchino which is a soft cheese produced in the
Northern Italy (Pepe et al., 2016). More than 800 peptides were reported
to be released during the 12 months processing of Parma cheese. From them,
18 were reported as ACE inhibitors like VPP, IPP, LHLPLP, HLPLP, or
ENLLRF and 12 longer peptides (>12 amino acids) as antimicrobial
(Solieri et al., 2020). ACE inhibitory peptides VPP and IPP were observed
to increase up to 18 months of ripening of Parma cheese and then decreased
at 24 months of ripening (Martini, Conte, & Tagliazucchi, 2020). Six pep-
tides were selected for DPP-IV and α-glucosidase inhibitory activity by
combining peptidomics, in silico analysis, and a structure–activity relation-
ship, outstanding peptide APFPE as a powerful DPP IV inhibitor
(Martini, Solieri, Cattivelli, et al., 2021; Martini, Solieri, & Tagliazucchi,
2021). Furthermore, ACE inhibitory peptides HLPLP and LHLPLP were
demonstrated to be resistant to simulated gastrointestinal digestion and able
to be partially absorbed during transepithelial transport in a Caco-2 cell
monolayer (Basiricò et al., 2015).
6. Release of bioactive peptides through food proteins

hydrolysis
The activity of endogenous enzymes is quite limited and depends on
the time of processing and conditions found in the food (pH, moisture
content, temperature, redox potential) during the process. This usually

results in low yields of peptides for much foods. In this way, when the objec-
tive is the large production of bioactive peptides, the procedure is to hydro-
lyze food proteins with exogenous peptidases or proteolytic microorganisms
(see Fig. 16). Numerous peptidases are commercially available as shown in
Table 2. Mostly used peptidases in commercial production are Alcalase from
Bacillus licheniformis, Protamex from Bacillus sp., Thermolysin from Bacillus
stearothermophilus, Flavorzyme from Aspergillus oryzae, Prolidase from
Lactobacillus casei, Neutrase from Bacillus subtilis or Bacillus amyloliquefaciens,
among other (Toldrá et al., 2020a). Each enzyme has its specific active site
and mode of action and therefore the selected enzyme, the protein source
used as substrate and the degree of hydrolysis will determine the obtained
yield of peptides and their bioactivity (Xing, Li, Toldrá, & Zhang, 2021).
Some variations in protein hydrolysis may be due to altered enzyme
reproducibility and stability as a consequence of batch to batch variability
(Merz, Apple, et al., 2016; Toldrá et al., 2018).
Commercial peptidases may exert some side activities in addition to the
main enzyme activity shown in the manufacturers specifications and
this may affect the peptides pattern as well as the hydrolysis degree. For
instance, it has been reported that Flavourzyme, an enzyme extracted from
Aspergillus oryzae, was characterized to have a wide activity spectrum con-
sisting of three endopeptidases, two dipeptidylpeptidases, two aminopepti-
dases and one amylase (Merz et al., 2015; Merz, Apple, et al., 2016).
Furthermore, aminopeptidase and carboxypeptidase activity, based on the
release of free amino acids, was reported for other enzymes like Alcalase
2.4 L (Novozymes), Maxazyme NNP DS (DSM), Flavourzyme 1000 L
(Novozymes) and Protease AN (Amano Enzyme Inc.) while a major endo-
peptidase activity was reported for Bioprase SP-20FG (Nagase), Collupulin
200 L (DSM), Corolase2TS (AB Enzymes), Promod 439 L (Biocatalysts
Ltd.), Proteinase T (DuPont) and Protin SD-AY10 (Amano Enzyme
Inc.) (Merz, Claaßena, et al., 2016).
A large number of bioactive peptides generated through enzymatic
hydrolysis of food proteins, usually isolated from food waste and food
co-products have been reported in the literature and some examples are
given in Table 3. For instance, protein rich extract from almond flour
was hydrolyzed with a neutral protease from Bacillus subtilis and 208 small
bioactive peptides with 2–4 amino acids sequences were identified
(Huang et al., 2022). Some of the most relevant were reported to be VY
and FY, both dipeptides as ACE inhibitors with in vivo hypotensive activity
in animal assays (Suetsuna, Maekawa, & Chen, 2004), FK has also antioxi-
dant activity (Huang et al., 2022). Dipeptides GW, VF and YP found in
Table 2 Commercial enzyme preparations with specific characteristics and some relevant applications.
Commercial
preparation Origin Manufacturers Activity Cleavage sites Application References
Flavourzyme Aspergillus oryzae 3 endopeptidases Cereals Huang et al. (2015),
1000 M 2 aminopeptidases Calcium chelating Meinlschmidt,
2 peptides, Soy Sussmann,
dipeptidylpeptidases Schweiggert-Weisz,
1 α-amylase and Eisner (2016),
Merz et al. (2015)
Valkerase Bacillus licheniformis Bri Enzymes Keratinase, Serin Non especific Feather meal –
Endopeptidase
Prolidase L-lactis cremoris Dipeptidase Bonds including proline Cheese making Kitchener and Grunden
Other many sources or hydroxiproline (2012)
Bioprase Bacillus sp. Subtilisin Merz et al. (2015)
SP-20FG Endo
metalloprotease
Aminopeptidase
Neutrase Bacillus subtilis Novozymes Metalloprotease Collagen Ou et al. (2010),
B. amyloliquefaciens Calcium- and Meinlschmidt et al.,
iron-chelating 2016
peptides Soy
Alcalase 2.4 L Bacillus licheniformis Novozymes Subtilisin Non especific Calcium-chelating Choi, Lee, Chun, and
Alkaline serin peptides Song (2012),
endopeptidase Charoenphun,
Extracellular neutral Cheirsilp, Sirinupong,
metallo protease and Youravong (2013)
Aminopeptidase
Savinase Bacillus lentus. Novozymes Subtilisin Higher affinity by Phe Lentil proteins Garcı́a-Mora, Peñas,
Alkaline serin and Leu Frias, and
endopeptidase Martı́nez-Villaluenga
(2014)
Esperase Bacillus lentus. Novozymes Subtilisin Broad especificity Georgieva, Stoeva,
Alkaline serin Voelter, Genov, and
endopeptidase Betzel (2001)
Protamex Bacillus licheniformis Novozymes Subtilisin Calcium- chelating Sung et al. (2014),
Bacillus Serin peptides Slizyte et al. (2016),
amyloliquefaciens endopeptidase, Bioactivity in Fish, Meinlschmidt et al.
Metallo Antiallergenic in (2016)
endopeptidase cereals
Neutral protease Soy
Protex 6 L Bacillus licheniformis Genencor Alcaline serine Bioactivity in Fish Slizyte et al. (2016)
endopeptidase
Protease N-01 Bacillus subtilis ASA Endoprotease
Spezialenzyme
GmbH
Protease M Endo & Exo Calcium chelating Lv, Bao, Liu, Ren, and
peptides in soybean Guo (2013)
Promod 439 L Bacillus licheniformis Biocatalysts Subtilisin Generation of flavor Mihulová, Vejlupková,
in animal, vegetable Hanušová, Štětina, and
and fish proteins. Panovská (2013)
Pronase Streptomyces griseus Sigma-Aldrich Endo & Exo Yoshida et al. (1988)
Corolase 7089 Bacillus Subtilis AB Enzymes Neutral Bioactivity in Fish Garcı́a-Mora, Peñas,
GmbH Endopeptidase Frias, and
Martı́nez-Villaluenga
(2014), Slizyte et al.
(2016)
Continued
Table 2 Commercial enzyme preparations with specific characteristics and some relevant applications.—cont’d
Commercial
preparation Origin Manufacturers Activity Cleavage sites Application References
Corolase PP Porcine pancreatic AB Enzymes Endopeptidase, Bioactivity in Fish Slizyte et al. (2016)
gland GmbH Amino- &
carboxypeptidase
Corolase 2TS Bacillus AB Enzymes Endopeptidase Cereals, soy Merz et al. (2015),
thermoproteolyticus Meinlschmidt et al.
Bacillus (2016)
stearothermophylus
GC710 Genencor Neutral proteinase Cereals
Biotech
GC106 Aspergillus niger Genencor Acid proteinase Antiallergenic in Sung et al. (2014)
Biotech Aspartic-type cereals
peptidase
Pancreatic Porcine pancreatic Novozymes Endopeptidase Meinlschmidt et al.
trypsin Novo glands (2016)
Trypsin Bovine pancreas Sigma-Aldrich Endopeptidase Broad specificity Calcium-chelating Guo et al. (2015)
Fluka Arg-j-Xaa, Lys-j-Xaa peptides from shrimp
by-products, Tilapia
or Alaska Pollock skin
Chymotrypsin A Bovine pancreas Sigma-Aldrich Endopeptidase Tyr-j-Xaa, Trp-j-Xaa,
Fluka Phe-j-Xaa, Leu-j-Xaa
Chymotrypsin C Bovine pancreas Sigma-Aldrich Endopeptidase Leu-j-Xaa, Tyr-j-Xaa,
Fluka Phe-j-Xaa, Met-j-Xaa,
Trp-j-Xaa, Gln-j-Xaa,
Asn-j-Xaa
Pepsin Porcine Gastric Sigma-Aldrich Hydrophobic,
mucosa Fluka preferably aromatic,
residues in P1 and P1’
positions
Alkaline protease Bacillus licheniformis Genencor Serine-type Aromatic or Antiallergenic in Sung et al. (2014)
peptidase hydrophobic residues cereals
Seabzyme L 200 Carica papaya Speciality Endoprotease Hydrolysis of proteins
Enzymes & with broad specificity
Biotechnologies for peptide bonds, but
preference for an amino
acid bearing a large
hydrophobic side chain
at the P2 position
Bromelain Pineapple stem Great food Cysteine-type Broad especificity Antiallergenic in Sung et al. (2014)
(Biochem) peptidase cereals
Collupulin Carica papaya Gist-brocades Cysteine-type Aromatic or Antiallergenic in Sung et al. (2014)
peptidase hydrophobic residues cereals
Papain Carica papaya Sigma-Aldrich Cysteine-type Broad especificity Collagen Sung et al. (2014)
Fluka peptidase Antiallergenic in
cereals
Ficain (ficin) Figs latex Sigma-Aldrich Cysteine-type Broad especificity
Fluka peptidase
Xaa: chain with amino acids at the C-terminal.
Reproduced from Toldrá, F., Reig, M., Aristoy, M.C., Mora, L. (2018). Generation of bioactive peptides during food processing. Food Chemistry, 267, 395–404. Generation of bioactive
peptides during food processing. Food Chemistry, 267, 395–404, with permission from Elsevier.
Table 3 Examples of bioactive peptides released by hydrolysis with commercial peptidases under controlled conditions.
Food Type Treatment hydrolysis Peptide sequence Potential activity References
Almond Flour Neutral protease Bacillus subtilis VY, FY ACE inhibitory Huang, Goncalves
(0.5%, 50 °C, pH 9, 2 h) Dias, de Moura, Bell,
and Barile (2022)
Milk Goat (Capra Trypsin (3%, 37 °C, pH 8, 3 h) INNQFLPYPY, Antidiabetic Zhang, Chen, Ma, and
hircus) milk SPTVMFPPQSVL Chen (2015)
MHQPPQPL
Algae Gracilariopsis Trypsin (2%, 2 h) FQIN[M(O)]CILR, TGAPCR ACE inhibitory Deng et al. (2018)
lemaneiformis
(Rhodophyta)
Palmaria palmata Corolase PP (2%, 50 °C, pH 7, 4 h) SDITRPGGQM Antioxidant Harnedy, O’Keeffe,
and FitzGerald (2017)
Red seeweed Pepsin (1%, 37 °C, pH 2, 3 h) GGSK, ELS Antidiabetic Admassu, Gasmalla,
(Porphyra spp) Yang, and Zhao
(2018)
Spirulina platensis Pepsin (6%, 37 °C, pH 2, 10 h) CANPHELPNK, NPVWKRK, Anti-obesity Fan, Cui, Zhang, and
NALKCCHSCPA, Zhang (2018)
LNNPSVCDCDCMMKAAR
Fish Atlantic salmon Corolase PP (1%, 50 °C, pH 7, 1 h) GPAV, FF ACE inhibitory Neves, Harnedy,
(Salmo salar) Antidiabetic O’Keeffe, and
Antioxidant FitzGerald (2017)
Cuttlefish (Sepia Bacillus mojavensis (3 U/mg, 50 °C, AFVGYVLP ACE inhibitory Balti et al. (2015)
officinalis) pH 10)
Cuttlefish hepatopancreas enzymes EKSYELP, VELYP ACE inhibitory Balti et al. (2015)
(3 U/mg, 50 °C, pH 8)
Leatherjacket Insoluble bromelain (0.5%, 50 °C, AER, EQIDNLQ ACE inhibitory Salampessy, Reddy,
(Meuchenia sp.) 2 h) Phillips, and
Kailasapathy (2017)
Insoluble papain (0.5%, 50 °C, 6 h) DPHI, EPLYV ACE inhibitory Salampessy et al.
(2017)
Insoluble flavourzyme (1.25%, WDDME ACE inhibitory Salampessy et al.
50 °C, 2 h) (2017)
Sardinelle Bacillus amyloliquefaciens (4%, 37 °C, ITALAPSTM, SLEAQAEKY ACE inhibitory, Jemil et al. (2016)
(Sardinella aurita) 24 h) Antioxidant
GTEDELDKY Antioxidant Jemil et al. (2016)
Bacillus subtilis (4%, 37 °C, 24 h) NVPVYEGY Antihypertensive, Jemil et al. (2016)
Antioxidant
Smooth hound Esperase (50 °C, pH 9, 9.5 h) IAGPPGSAGPAG, ACE inhibitory Abdelhedi et al. (2016)
viscera VVPFEGAV, PLPKRE
Pacific herring Trypsin (1.39 U/Kg, 32.06 °C, pH KEEKFE, LHDELT Antioxidant Wang et al. (2019)
(Clupea pallasii) 6.78, 7 h)
Legumes Soy Alkaline proteinase (6 U/Kg, 50 °C, LLPLPVLK, SWLRL, WLRL Antidiabetic Wang et al. (2019)
pH 9)
Erythrina edulis Alcalase (0.5%, 50 °C, pH 8.3, 2 h) CCGDYY, DGLGYY, Antioxidant Intiquilla et al. (2018)
(pajuro) GESWCR, YDLHGY,
SQLPGW, WAL
FLWGKSY, WPW, Antiinflammatory Intiquilla et al. (2018)
SFMNVKHWPW
Duck (Anas Protamex (0.75%, 50 °C, pH 6, 4 h) AGRDLTDYLMKIL Antioxidant
platyrhynchos) GYDLGEAEFARIM
Continued
Table 3 Examples of bioactive peptides released by hydrolysis with commercial peptidases under controlled conditions.—cont’d
Food Type Treatment hydrolysis Peptide sequence Potential activity References
LQAEVEELRAALE Wang, Huang, Chen,
NWDDMEK Huang, and Zhou
IEDPFDQDDWGAWKK (2015)
Kacang goat Protamex + Flavourzyme (0.5%, FQPS ACE inhibitory Mirdhayati,
(Capra aegagrus 50 °C, pH 7, 4 h) Hermanianto, Wijaya,
hircus) Sajuthi, and Arihara
(2016)
Pork loin Thermolysin (0.008%, 5 °C, 24 h) LVGRPRHGQ, VFPS ACE inhibitory Choe et al. (2019)
By- Chicken combs Alcalase (5%, 4 h) APGLPGPR, FPGPPGP, Piro- ACE inhibitory Bezerra et al. (2019)
products and wattles GPPGPT
Bovine gelatin Thermolysin (65 °C, pH 8, 6 h) AG, AGP, VGP, PY, QY, DY, ACE inhibitory Herregods et al. (2011)
IY
Oil palm (Elaeis Alcalase (0.5%, 45 °C, pH 8.5, 2 h) ADVFNPR, VIEPR, LPILR, ACE inhibitory Zheng, Li, Zhang,
guineensis Jacq) + flavourzyme (0.5%, 50 °C, pH 7, VVLYK Ruan, and Zhang
kernel expeller 2 h) + pepsin (0.3%, 37 °C, pH 2, (2017)
1 h) + trypsin (0.3%, 37 °C, pH 7,
1 h)
Tomato seeds Bacillus subtilis (2%, 37 °C, 24 h) DGVVYY ACE inhibitory Moayedi et al. (2018)
GQVPP Antioxidant Moayedi et al. (2018)
Another random document with
no related content on Scribd:
yet tempting often to extravagance in those weeks when it is high; a
twelve-year limit permitted by the child labor law, and adult wages
that necessitate the children’s going to work as soon as that law
allows; the father rarely earning much more, and sometimes even
less, than the younger members of the family; scant amusement,
usually only the moving picture show, possible on the meager
income; poor health with the doctor often an impossible luxury.
JOTTINGS
DETROIT BOOSTING FOR SAFETY
The campaign for safety is taking firm root in Detroit. The Detroit
Manufacturers’ Association has in its employ two safety inspectors
who are at the call of members for work in their plants at any time.
They are constantly hunting for danger points and suggesting
methods of eliminating them.
More recently, following the enactment of the Workmen’s
Compensation Law, there has been organized the Detroit Accident
Prevention Conference. There have been three meetings so far, with
such men as John Calder of the Cadillac Motor Car Company and W.
H. Bradshaw, safety director of the New York Central lines as
speakers and papers by those members who were equipped by
reason of experience to give instructive information. The meetings
are held in the evening in a down town hotel where a moderate
priced dinner is served, the addresses and discussions following. The
average attendance has been about one hundred. As no membership
fee is charged and as great enthusiasm is displayed it is hoped that
shortly the attendance will be double this number.
TRADE SCHOOL FOR PRINTERS
In Printing Trade News the recently established School for

Printers’ Apprentices in New York is described by A. L. Blue, director
of the school. The school is co-operative in the extreme; it is
managed by a joint committee of employers (The Printers’ League),
workmen (the New York Typographical Union) and the public (the
Hudson Guild). Its headquarters are at the guild. The courses, which
are for working apprentices, are so planned as to develop
individuality. Afternoon classes are held for boys employed on the
morning papers, evening classes for others. The present enrolment is
ninety-six.
SICKNESS INSURANCE IN WISCONSIN
A bill marking the initial step towards the establishment of state

accident and sick benefit insurance is pending in the Legislature of
Wisconsin. This is one of the first proposals of the kind submitted in
any state. Its insurance features are modelled after the English act.
The bill applies solely to vocational diseases. Both employer and
employe are to contribute toward the premiums. Single employes
earning less than $600 a year, who have someone dependent upon
them, are eligible to protection under the provisions of the bill; no
person may come under its terms who earns over $900. Persons
earning $800 a year must have two dependent upon them, and those
earning $900 annually must have four persons dependent upon
them in order to come within the proposed statute.
Employers are to be allowed to deduct 1 per cent of the wages of
employes and they must add to this sum one-half of 1 per cent of the
pay roll, the entire sum to be paid into a state insurance fund. When
ill, the employe is to receive 65 per cent of his wages during the
period of his illness, but for not more than twenty-six consecutive
weeks nor more than thirty-nine weeks in a single year. If the
employe is sent to a hospital, his regular wages are to be paid to him
weekly. The State Industrial Commission is empowered to enforce
the provisions of the act in the event of its passage.
MUNICIPAL MINIMUM WAGE
A minimum wage of 25s. ($6.08) a week for all able-bodied men

will henceforth rule, says Life and Labor, in the municipal service in
Glasgow. It is now many years since the corporation of Glasgow
acknowledged the principle of a minimum wage, the rate then
introduced being 21s. ($5.11). Since that time improvements have
brought the wages up to an average minimum of about 23s. ($5.60).
so that the proposal for a minimum of 25s., which was carried in the
town council, means an advance of about 2s. ($0.48 2–3) weekly to
many of the lower-paid workmen. To give effect to the proposal an
additional expenditure of $41,365 will, it is estimated, be involved.
The position in Manchester is better, from the workers’ point of
view, than it will be in Glasgow even when the minimum weekly
wage is raised to 25s. ($6.08). Seven years ago the Manchester city
council raised the minimum wage to 25s. Early in the present year
there was an agitation for an increase of 2s. ($0.48 2–3) a week in
view of the increased cost of living. A special committee reported in
favor of an advance to 26s. ($6.33) a week, and this the council
agreed to. This sum is paid to all the laborers (as distinct from skilled
workers in the several departments) throughout the city.
FULL CREW BILLS
An unusual publicity campaign on the part of railroads has

resulted from the passage by the state Legislatures of the so-called
Full Crew Bills in New Jersey and New York, regulating the number
of employes on trains. In the New York newspapers for several days
in succession the railroads used three-quarter page advertisements
for a joint statement of their opposition. In this space they urged the
governor to veto the bill, and the public to protest against its
enactment. It is claimed by the railroads that the law will cost them
$2,000,000 annually in the state of New York without bringing any
increase in efficiency or safety. They point out that Governors
Hughes and Dix both refused to approve similar measures on the
ground that such questions should logically be decided by the Public
Service Commission.
In their advertisements the railroads urged that the matter be left
to the state Public Service Commissions, and promised to abide by
their decisions.
The Brotherhood of Railway Trainmen, which is urging such
legislation all over the country, insists that it is necessary to promote
safety. The Railroad Trainman, organ of the brotherhood says:
“Today our men are asking for legislation that is no more of a
departure from the beaten path than the safety device legislation of
twenty years ago was. They have tried to regulate the car limit of
trains and the number of men to be employed on them through their
contracts. They have failed in the first instance altogether and for the
most part in the other. They realize that, operated as trains are,
freight train service is often performed under unsafe conditions. Two
men for an unlimited number of cars is the rule for the most part.
Because of it there are freight trains running today averaging
between fifty and one hundred and thirty-five cars and two men are
in charge with the conductor.
“There will be trains, perhaps, on which the extra man will not be
needed, but if the companies had been forehanded enough to put
men where they were needed they could have saved the ones not
needed, but they did not and legislation does not find a way to
discriminate as readily as the exercise of common sense does.”
The bills have been signed and have become laws in both New
Jersey and New York.
CHURCH AND COMMUNITY
Edited by GRAHAM TAYLOR
TERRE HAUTE’S LABOR PARLIAMENT

BENJAMIN B. TOWNE
Failure on the part of the churches of Terre Haute, Ind., to grasp
the problems of its 11,000 workingmen led to the holding of a “labor
parliament.” This parliament, convened last May, was directed by
Harry F. Ward of the Methodist Federation for Social Service. There
were three meetings in different churches, where the problems of
industry and Christianity were discussed in an open and frank
manner.
But the prime movers realized, early in April, that to make this
parliament a success much local work would have to be done. As a
stepping-stone, the ministers adopted an industrial creed, which was
floated over the city, with the result that the laboring man discovered
that he and the church had common ideals toward which to aim.
The local work in the churches was adapted to the particular
condition of the locality, all efforts, however, being focused on the
labor parliament to be held in May. Shop meetings were held,
lantern slides of existing conditions were shown, and mass meetings
for working men and girls conducted. Besides these features, the
newspapers helped this most interesting scheme along, so that by the
time set for the labor parliament, all Terre Haute was prepared for
the co-operative discussion, which was to prove so beneficial to the
church and organized labor. The Central Labor Union co-operated
well with the movement and appointed a committee of three
prominent labor men to help the ministerial committee.
The labor parliament was, indeed, a success. Dr. Ward chose as his
subjects, Industry and Social Waste, Democracy in Industry, and the
Industrial Problem of Christianity. In all his talks Dr. Ward opened
the eyes of labor world and church. One, he showed, could not be of
full benefit in its community without the co-operation of the other.
And now, nearly a year after this industrial revival, what are the
results? Are any permanent effects apparent from these efforts, or
did the movement, swelling into the three days’ parliament,
gradually fade away and become forgotten by the laboring man? A
few pointed statements of those nearest the problem of both the
church and laboring man will show the result.
A. M. Powers, president of the Central Labor Union, has this to say
of its success. “The movement has been beneficial, as far as I can see,
to both sides. When the church can show that the laboring man is not
an insect to be placed upon a sociological dissecting table for amused
speculations of theologians, but a man to be helped and to help
advance the cause of the brotherhood of man through the church,
then the antagonism will be replaced by a hearty co-operation
because this spirit of brotherhood is the basis of the organized labor
movement.
“I believe the churches of Terre Haute have shown that this is the
spirit of their activity in their last year’s efforts, and as an individual I
endorse the movement and think that as long as the same spirit is
shown the labor unions will be willing to work hand in hand with the
church.”
George W. Greenleaf, secretary-treasurer of District Lodge No. 72,

International Association of Machinists, and city councilman, says:
“The labor parliament and the preceding church services held in Terre Haute last
winter were beyond the question of a doubt a benefit to organized labor. The chief
benefit derived, in my estimation, consisted in the dispelling of the popular
prejudice against our organizations and the placing of our cause on a higher plane
in the minds of the public.”
Terre Haute’s Industrial Creed

United we stand:
For equal rights and perfect justice to all men.
For the principle of conciliation and arbitration.
For the protection of workers from dangerous machinery, occupational
diseases, injuries and mortality.
For the abolition of child labor.
For such regulations of conditions of labor for women as shall safeguard
the physical and moral health of the community.
For the suppression of “the sweating system.”
For a reasonable reduction of the hours of labor to the lowest practical
point with labor for all and a reasonable degree of leisure.
For release from employment one day in seven, and whenever at all
possible that this be the Sabbath Day.
For the highest wage that each industry can afford and for the most
equitable division of the profits of industry that can be devised.
For the recognition of the Golden Rule (Matt. 7:12) and the teachings of
Christ as the supreme law of society and the sure remedy of all ills.
The ministers of the city feel much the same way about the effects
of the parliament.
Rev. A. E. Monger, pastor of the largest Methodist church in the
city and one of the promoters of the movement, says:
“Since the campaign there has been crystalized in the churches a sentiment of
responsibility for the welfare of the laboring man. The laboring men have found
that the gospel does have a message against the great sins under which they are
struggling.”
As a further evidence of the parliament’s lasting effect, Rev. John
G. Benson, another of its promoters, may be quoted:
“We are getting requests from every quarter for a repetition of the parliament.”
NEW RECOGNITION OF SOCIAL
CHRISTIANITY
In religious periodical literature two high notes of social
significance have recently been struck. The Constructive Quarterly
has appeared from the press of the George H. Doran Company in
America and Hodder & Stoughton in England. It is planned to be a
free forum where all the churches of Christendom may frankly and
fully state their “operative beliefs” and their distinctive work,
“including and not avoiding differences,” but making “no attack with
polemical animus on others.”
The purpose of this undertaking is to afford opportunity for the
churches, without compromise, “to re-introduce themselves to one
another through the things they themselves positively hold to be vital
to Christianity,” “so that all may know what the differences are and
what they stand for, and that all may respect them, in order to
cherish and preserve whatever is true and helpful and to discover
and grow out of whatever is harmful and false.”
As it has no editorial pronouncements and no scheme for the unity
of Christendom to promote, the Quarterly will depend upon the
catholicity and representative influence of its editorial board,
selected from all countries and communions, to promote a fellowship
of work and spirit. The middle term of the Quarterly’s subtitle—a
journal of the Faith, Work and Thought of Christendom—is likely to
prove the basis for the correlation of the other two. For long before
the faith and the thought of Christendom may be correlated, the
churches will surely co-operate in their common work.
The Hibbert Journal, which for ten years has been the ablest
technical quarterly review of theology and philosophy, announces a
department of social service. This policy was foreshadowed by the
editor as early as October, 1906, in a notably direct and able protest
against the church standing aloof from “the world.” He stoutly
maintained that
“the alienation from church life of so much that is good in modern culture, and so
much that is earnest in every class, is the natural sequel to the traditional attitude
of the church to the world.”
How false and unintelligible, as well as untenable, this attitude is
appears in these categorical imperatives:
“If by ‘the world’ we mean such things as parliamentary or municipal
government, the great industries of the nation, the professions of medicine, law,
and arms, the fine arts, the courts of justice, the hospitals, the enterprises of
education, the pursuit of physical science and its application to the arts of life, the
domestic economy of millions of homes, the daily work of all the toilers—if, in
short, we include that huge complex of secular activities which keeps the world up
from hour to hour, and society as a going concern—then the churches which stand
apart and describe all this as morally bankrupt are simply advertising themselves
as the occupiers of a position as mischievous as it is false.
“If, on the other hand, we exclude these things from our definition, what, in
reason, do we mean by ‘the world?’ Or shall we so frame the definition as to ensure
beforehand that all the bad elements belong to the world, and all the good to the
church? Or, again, shall we take refuge in the customary remark that whatever is
best in these secular activities is the product of Christian influence and teaching in
the past? This course, attractive though it seems, is the most fatal of all. For if the
world has already absorbed so much of the best the churches have to offer, how can
these persist in declaring that the former is morally bankrupt?
“Extremists have not yet perceived how disastrously this dualistic theory thus
recoils upon the cause they would defend. The church in her theory has stood aloof
from the world. And now the world takes deadly revenge by maintaining the
position assigned her and standing aloof from the church.”
No better prospectus for the social work of either of these great
quarterlies could be framed than the intention to demonstrate and
bear home to the intelligence, conscience and heart of the churches
these very affirmations. For, while enough of church leaders and
followers thus face forward to warrant Professor Rauschenbusch in
declaring that it has at last become orthodox to demand the social
application of Christianity, yet there is a sharp reaction within every
denomination, which threatens to retard this hopeful movement of
the churches to serve their communities and thereby save
themselves.
But the ultimate issue between those who are thus fearlessly facing
the present and those who persist in backing up into the future
cannot be doubtful. Social Christianity is not only demonstrably
orthodox, but has won its recognition and its own place in any
theological, philosophical, historical or experiential conception of
Christianity that claims to be comprehensive, not to say intelligent.
Without a much larger emphasis upon the social aims and efforts of
Christianity in the thought, belief and work of the church, the need
that is finding expression in every parish and community cannot be
met—that which the Constructive Quarterly well states to be “the
need of the impact of the whole of Christianity on the race.”
THE FIRST ORPHAN ASYLUM IN THE
[8]
UNITED STATES
THAT OF THE URSULINE NUNS AT NEW
ORLEANS
8. This account of the founding of our first orphanage in the quaint language
of the time was obtained for The Survey from a friend of the institution by Albert
H. Yoder.
At the outset of the colonization of Louisiana by the French, ten
Ursuline nuns of France, with noble generosity and self-sacrifice,
volunteered to go to New Orleans, there to instruct the children of
the colonists. They left Rouen in January, 1727.
After great difficulties and countless perils, they reached the
mouth of the Mississippi whose waters they ascended in pirogues.
They finally landed in the Crescent City on the morning of August 7,
1727, after a sea voyage of nearly six months. They had set sail from
the port of Havre on February 23, 1727 after a month spent in Paris.
Arriving in New Orleans, they were met by Bienville, governor of
the province of Louisiana. As there were no proper accommodations
yet provided, the governor vacated his own residence and placed it at
their disposal for a convent and school. Immediately was begun the
erection of a new building which was completed in 1734.
The Ursuline nuns upon its completion took possession and
occupied it till 1824 when they removed to their present home below
the city. This structure, which is now the Archbishopric, or official
place for the transaction of the business of the Archdiocese of New
Orleans, is the oldest building in Louisiana and also in the vast
extent of what was known as the Louisiana Purchase.
The Ursulines began their self-sacrificing work immediately upon
their arrival on August 8, 1727 and opened a free school to which
were added a select boarding school and then a little later a hospital.
Moreover, in order to inculcate principles of civilization and,
especially, of religion in the hearts of the wives and daughters of the
Negroes and Indians, the nuns devoted one hour each day to their
instruction.
Shortly after their arrival a new field of labor was open to their zeal
in the shape of a poor orphan whom Father de Beaubois, had
withdrawn from a family of dissolute morals. Although their lodgings
at the time were insufficient, the nuns being still in Bienville’s house
(their new convent, the present old Archbishopric, was not ready for
occupancy until July 17, 1734), they adopted the child. This was the
tiny mustard-seed from which sprang the flourishing orphanage
which exists to the present day. It proved a real providence for the
country, especially in colonial times, as may be gleaned from
history’s record of the Natchez massacre, which took place on
November 28, 1729.
After this frightful tragedy, so pathetically described by
Chateaubriand, the Indians, who had spared only the young wives
and daughters of their French victims, were forced to give up their
hostages or to be massacred in turn. The generous Ursulines then
opened their home to these unfortunate little ones and mothered
them.
This act of disinterestedness and charity was truly heroic,
considering the great difficulties usually attendant on the founding of
a colony and was highly commended by Rev. Father le Petit, Jesuit,
in a letter addressed, July 12, 1730, to Rev. Father d’ Avaugour,
procurator of the American missions. Having given an account of the
appalling massacre of the French at Fort Rosalie by the Natchez
Indians, Rev. Father le Petit adds:
“The little girls, whom none of the inhabitants wished to adopt, have greatly
enlarged the interesting company of orphans whom the religieuses [Ursulines] are
bringing up. The great number of these children serves but to increase the charity
and the delicate attentions of the good nuns. They have been formed into a
separate class of which two teachers have charge.
“There is not one of this holy community that would not be delighted at having
crossed the ocean, were she to do no other good save that of preserving these
children in their innocence, and of giving a polite and Christian education to young
French girls who were in danger of being little better raised than slaves. The hope
is held out to these holy religieuses that, ere the end of the year, they will occupy
the new house which is destined for them, and for which they have long been
sighing. When they shall be settled there, to the instruction of the boarders, the
orphans, the day scholars, and the Negresses, they will add also the care of the sick
in the hospital, and of a house of refuge for women of questionable character.
Perhaps later on they will even be able to aid in affording regularly, each year, the
retreat to a large number of ladies, according to the taste with which we have
inspired them.
“So many works of charity would, in France, suffice to occupy several
communities and different institutions. But what cannot a great zeal effect? These
various labors do not at all startle seven Ursulines; and they rely upon being able,
with the help of God’s grace, to sustain them without detriment to the religious
observance of their rules. As for me, I fear that, if some assistance does not arrive,
they will sink under the weight of so much fatigue. Those who, before knowing
them, used to say they were coming too soon and in too great a number, have
entirely changed their views and their language; witnesses of their edifying conduct
and great services which they render to the colony, they find that they have arrived
soon enough, and that there could not be too many of the same virtue and the same
merit.”
After giving details relative to the visit of the Illinois chiefs, who
had come to condole with the French and to offer help against the
Natchez, Father Le Petit adds:
“The first day that the Illinois saw the religieuses, Mamantouenza, perceiving
near them a group of little girls, remarked: ‘I see, indeed, that you are not
religieuses without an object.’ He meant to say that they were not solitaries,
laboring only for their own perfection. ‘You are,’ he added, ‘like the black robes,
our fathers; you labor for others. Ah! if we had above there two or three of your
number, our wives and daughters would have more sense.’ ‘Choose those whom
you wish.’ ‘It is not for me to choose,’ said Mamantouenza. ‘It is for you who know
them. The choice ought to fall on those who are most attached to God, and who
love him most....’”
The records make mention of Therese Lardas, daughter of a
Mobile surgeon. After her father’s death, her mother brought her to
the Ursuline orphanage, where she intended leaving her just long
enough to make her first communion; but, when she came to take
her home, so earnestly did the child plead to remain, that the mother
could not resist her entreaties. At the age of sixteen, she entered the
novitiate. She led the life of an exemplary lay sister, and died at the
age of twenty-nine on November 22, 1786.
In testimony of the good education given to all classes by the
Ursulines, the Rt. Rev. Luis Penalvery Cardemas said in a dispatch
forwarded to the Spanish court, November 1, 1795:
“Since my arrival in this town, on July 17, I have been studying with the keenest
attention the character of its inhabitants, in order to regulate my ecclesiastical
government in accordance with the information which I may obtain on this
important subject.... Excellent results are obtained from the Convent of the
Ursulines, in which a good many young girls are educated. This is the nursery of
those future matrons who will inculcate in their children the principles which they
here imbibe. The education which they receive in this institution is the cause of
their being less vicious than the other sex....”
Up to 1824, that is, for well nigh a century, the Ursulines
maintained their orphanage in what is now the old Archbishopric. At
this period, New Orleans having spread considerably and become too
densely populated to afford the advantages and charms of the
country so necessary to a large boarding school, the institution was
removed three miles lower down, to the magnificent place which the
Ursulines hold to the present day. Owing to the encroachments of
the great Father of Waters, they are to transfer again, within a year,
to another site.
After 1824, several asylums having been founded for orphans of
both sexes, the Ursulines received but thirty or forty poor children.
In keeping with their sphere of life and future career, these children
are taught English, French, geography, arithmetic, elementary
history, and some housekeeping, sewing and laundry work. The nuns
endeavor, above all, by religions instruction and careful training, to
inculcate in the hearts and minds of their youthful charges principles
of duty, so as to form for the future women of confidence, courage,
self-sacrifice and devotion.
SOCIAL SERVICE OF THE PRESBYTERIAN
CHURCH IN CANADA
J. G. SHEARER
The Presbyterian church in Canada does social service work

through its Department of Social Service and Evangelism. Efforts are
directed along several lines.
Social surveys of both urban and rural communities are
conducted, considering not only religious and moral, but also social
and economic conditions. An expert is employed who gives all his
time to the work. He secures the co-operation of a large number of
volunteer helpers, many of whom are proficient in various phases of
social service work.
The problems of the city are studied and practical solutions
sought. This is attempted in the following ways:
By evangelical social settlements, of which there are one in
Montreal, one in Toronto and one in Winnipeg. Eight or ten others
in the not distant future are planned for various other growing
cities in the Dominion, especially where non-Anglo-Saxon
immigrants are numerous. Our organizer and supervisor of this
work is Sara Libby Carson, founder of Christodora House and
various other settlements in New York, St. Christopher House,
Toronto, and Chalmers House, Montreal. We also have established
a training school for settlement workers, in connection with St.
Christopher House, Toronto.
By securing the co-operation of churches and sympathetic
organizations in every variety of general social betterment effort.
By establishing special redemptive and social missions on the
crowded thoroughfares. The first of these was Evangel Hall,
Toronto, in which evangelistic work, as well as various sorts of
social work, is carried on.
The department has taken up in a large way redemptive and
preventive work in the interest of girls, and associated with that
educational work along the line of sex teaching among boys and men.
There are five homes which are called social service houses, in which
girls and women requiring special help are taken care of. Fifteen
trained Christian women give their time to this phase of the
department’s endeavor, and there is also a large army of volunteer
helpers. In connection with this work an educational campaign
through pulpit and platform and the distribution of literature
throughout the Dominion is carried on. From time to time
legislation, federal or provincial, for the more adequate protection of
girls and women is sought.
In co-operation with other interested bodies the department keeps
up a steady campaign for the suppression of gambling,
intemperance, sale of immoral literature, unclean theatricals, the
social vice, and the promotion of the positive virtues, the opposite of
these.
Special attention is being directed to positive effort and
constructive work along all lines aiming at social uplift, and a good
deal of legislation toward this end has been successfully put through.
The department has established a lantern slide and film service,
and is endeavoring to supply through illustrated means elevating
entertainment as well as information and inspiration.
All the evangelistic work of the Presbyterian church is done
through this department, so that evangelism and social service are
kept in close association in all effort undertaken.
SYNAGOGUE AND COMMUNITY
RABBI HORACE J. WOLF
Temple ‘Berith Kodesh’, Rochester, N. Y.
The changing relation of the synagogue and the community is

proving the truth of the hoary platitude that history repeats itself.
During the Middle Ages the synagogue was the heart of the secular as
well as of the religious life of the community; it was a social center as
well as a house of prayer. There the poor man found succor, the
stranger acquaintances, the children their teachers, and the young
people “their fates.” It would be almost impossible to list all the
private and public interests which, clustering about the synagogue,
bore witness to the vital part this institution played in medieval
Jewish life.
This prominent role was due to the enforced isolation of the
Jewish community; thanks to the Ghetto walls the Jewish group
constituted a city within a city. Once the Jewish population was
concentrated into separate quarters, the synagogue became to the
segregated community what the home was to the individual family; it
was not only a place of meeting, but also a clearing house for
individual and communal joys and sorrows.
But the intimacy was broken down by the political emancipation
that came to Jewry at the end of the eighteenth century. Slowly, as
the old functions of the synagogue were taken over by special
institutions housed in their own buildings, the synagogue began to
be used purely as a house of worship; aside from this, its sole
concern seemed to be the Sunday school. Applicants for charity were
referred to the charity office across the street; social functions took
place at the clubs; legal disputes were no longer decided by a
rabbinical court. True, there were few large cities in this country in
which the Jewish community did not point with pride to its
magnificent house of worship; but in the majority of cases these
gorgeous buildings (I am writing throughout of the synagogues of the
reform wing) were dark six days and nights a week. In this respect,
they differed little from the churches about them.
But the last decade, which has seen the rise of the institutional
church, is witnessing the return of the synagogue to its former close
relationship to communal Jewish life. The change is due to the same
causes that made for the broadening of the work of city churches.
The popular criterion of a social institution’s value, it was seen, is its
working efficiency. Men who judged by concrete and tangible
standards, and their number is legion, were becoming indifferent to
religion because it appeared divorced from life. The leaders of
American Judaism began to appreciate that it was insufficient to
proclaim from the pulpit that religion included charity, social
amelioration, good citizenship, as well as morality and reverence;
they began to insist that the synagogue should “monument its
claims.” It was urged that the synagogue should not only strive to
touch the religious nature of the people with the conventional
methods of prayer and praise and preachment, but should also bring
to bear a system of institutional activities, social, educational and
philanthropic which would bring it into contact with its members’
physical, mental and social nature as well.
As a result of this awakening there is hardly a synagogue in the
United States which has not some form of institutionalism—be it
only a sewing circle. A questionnaire sent out by the Committee on
Social and Religious Union of the Central Conference of American
Rabbis to its various members elicited ninety-seven replies. In these
answers seventy-one report the existence of congregational libraries;
eleven congregations conduct classes for the teaching of the English
language and instruction in citizenship; six maintain settlements;
two have labor bureaus; fifty list philanthropic activities, glee and
choral societies, athletic clubs, kindergartens, industrial schools and
dancing classes.
The committee in summarizing its report says: “The majority [of
our colleagues] feel that all these institutional creations have helped
to deepen the interest of the members in the synagogue and in each
other; that they have helped to make the temple a center for Jewish
communal life; ... that they religionize social functions; that they
stimulate the Jewish consciousness; that they prevent
disintegration....”
Once again the synagogue is playing a splendid role in Jewish
communal life. Men are beginning to perceive that the ideal
synagogue will be in use at practically all hours every day in the
week, will never be dark and deserted. The impressive appearing
edifice that was tenanted by silence and gloom on every day except
the Sabbath is becoming an anachronism. Our hope is that the
synagogues that continue to slumber may awaken before it is too
late, and take their proper share in the work of communal uplift.
DR. HOWARD KELLY’S APPEAL FOR
CHURCH CIVIC SERVICE
The demands for a better trained ministry and membership in the
churches are being strongly emphasized by such statements of what
the community expects of them as Dr. Howard A. Kelly of the Johns
Hopkins University medical faculty recently made in an address at
the annual meeting of the New York Probation and Protective
Association. In giving his consent to print some of his remarks, he
writes, with special reference to his efforts against the social evil:
“I feel as though my own work in this field were to bring the churches together
for neighborhood social interests. If we do not get the churches actively to work, I
believe all the social developments of the last thirty years are destined to failure. I
fully believe that a few strong men, say five or six in a city like Baltimore, can
effectively put persistent effort into the work of amalgamating our churches for the
expression of the Christian life in the active service of their fellow men.”
In his address in New York, after stoutly combating, from his
professional and public points of view, the policy of segregating vice,
he declared that the social work of the church is indispensable to
progress, and that it is the duty and the opportunity of the church to
fulfil the need in this direction. He spoke substantially as follows:
“The most effective of all agencies in breaking down the strongholds of vice and
in building up the national character is the church. For some reason unknown and
unfathomable, some of my associates in this beneficent work who don’t go to
church fight shy of discussing any enlistment of the churches everywhere. Not a
few who have never had any personal interests in the church even stand ready to
declare, with a distinguished head of our public libraries, that the church
represents the largest outlay of capital for the smallest return in interest the world
has ever seen.
“The utility of the church in the social field is best defended perhaps by citing an
investigation of over 1000 social workers of all kinds showing that over 90 per cent
are church people, and I venture confidently to affirm that if the inspiration of the
church direct and indirect is taken away from our various social movements, they
will die outright in short order. I can furthermore now aver what I could not have
said twenty years ago, of a group of splendid humanitarian workers who have no
church affiliations, that this indefatigable but weary band has at last come to
realize that unless the church comes to the front and does her duty this great
purifying work will never be done.
“The difficulty has been that our churches have been too much afflicted with
myopia, seeing little beyond the confines of their own four walls. They have also
one and all slipped into the easy ways of formalism, and worse still, the laity have
thrust the burden of their religious obligations onto the shoulders of a groaning,
overladen clergy, trusting to discharge their own personal responsibilities on a
cash basis by check. I am sure that the clergy are well aware that there is much to
be desired in the social relations of the church to the community and I believe no
set of men will show themselves more ready to advance on new lines if they can see
that the movement is really a spiritual one and that a large service can thus be
inaugurated.
“There are many reasons why the churches must be depended upon as the
backbone of any morals movement:
They are ideally distributed among the people.
They have the intelligence and the means.
They have a source of continuous inspiration needed in dealing with chronic
distressing problems.
They alone can guarantee perpetuity of effort.
“In utilizing the church, the minister must be the organizer and leader of his
people. A new relationship between pastor and layman will ensue, and laymen,
once drawn into a local work, will soon branch out into all forms of civic work for
the weal of the community. Again, the churches possess the community buildings
so much needed. The only other similar institution capable of a similar co-
operation on a large scale is the public school which, while valuable and necessary
in this movement, has not the independence and lacks the great inspiration.
“What, then, is the specific program for the church? First, of all, she must not
abate but rather increase her dependence upon God. She must never yield to
temptation to abandon the one really valuable quality she possesses by relegating
to the background the living fountains of inspiration she holds in God’s word, for a
mere mundane horizontal social Gospel which makes a religion of the human
activities which are but its appropriate outward expression. First a glance upward,
then outward to God for the life, and to the human arena for the sphere in which
the life must be manifested. This does not hinder but quickens the impulse to
effective service.
“The profounder my faith, the more am I able to work in affectionate association
and harmony with the many who do not see eye to eye with me here on earth; I
cannot, however, continue to work with any who demand as the price of their help
that I shall stifle all outward expression of my faith. He who walks in the light must
sing of the light lest the light he has shall fade into darkness, and he too shall be
left to flounder along the dead level of merely human self-guided impulses.
A PRAYER FOR EFFICIENCY

O God, as to an earthly father, we bring thee each our yearning confession
of failure to realize to the full the powers thou hast given us as laborers in
thy kingdom on earth. May we learn through this, our mutual prayer, to be
charitable to one another’s shortcoming. Teach us, by love if it may be, by
bitter rebellion, if it must be, that our prayer may be answered only as we
are firm to lend a hand in mutual aid and sympathy to the less fortunate.
Let each in strength supply his neighbors’ weakness, and build up in him the
efficiency which is his birthright.
Thus, in humility of heart, we pray for justice to our overstrained and
blighted brothers who never catch up, who grind their lives into sieves of
despair and deficit, each grist the harder because there is less of life to
spare. Think upon the handicapped in body and in soul, for whose
backwardness we are jointly responsible through our inefficiency. May we
give them health and leisure and knowledge and so joy and inspiration so
that, restored to themselves, they may in free good will repay them a
hundredfold, in deeds of brotherly gratitude and justice to others, for thy
sake.
And chiefly we pray for those in whom we have put our trust; that their
strength may be equal to the temptations of the power we have given them
from thee. May they realise that not their own gain, but social justice, must
measure the efficiency of their efforts. Bring home to their minds and hearts
the far-reaching power, for evil and for good, of industry and government,
of church and press; let them remember vividly the remote effects of
indifference and negligence in the web of modern life.
May the getters of gold give justice to its producers; may its earners have
charity toward its spenders; may the givers of gold be gifted with wisdom
and courage; and may all social workers feel the weight of an especial
responsibility; that the surplus wealth of which they are guardians may be
husbanded for its true purposes and not be betrayed, nor delayed, nor
wasted in their hands; that thou mayst have gratitude in turn toward all,
for thy children’s sake. Thus may thy kingdom grow on earth into fuller and
more abundant life for each and all.—AMEN.
“The church must be a great, perennial fountain of spiritual and moral energy to
the whole people in all the avenues of human interests. She must realize her
obligation to champion the cause of the oppressed, whatever the cause and
whoever the oppressor, whether in her fold or out of it. She must watch to prevent
the rich from grinding the faces of the poor. She must when necessary provide for
every legitimate desire of the people. If politics are corrupt, then she must enter
aggressively into the field of politics, only for purity and not for party. She must
fight all saloons and organize neighborhood opposition to their continuance, but
provide too for some form of social life to replace them.
“The rich churches most be big sisters to the poor, providing means and sending
talented workers wherever they are needed. If the church needs money for
neighborhood enterprise, let her lop off her choirs and stained glass windows and
bells, expensive altars, and put the money saved into human lives. She must
discourage all extravagances which give the poor just cause for bitterness and
arouse envy and set up unworthy standards. Let the church make a map of
neighborhood conditions. This will serve as an object lesson and as a basis for
action. In weekly classes she should then study such social problems as:
Social teachings in the Bible.

Tuberculosis in our city.
Prostitution.
Housing the poor.
Amusements.
Wages paid in department stores and factories.
Near town places of recreation.
Hotels, saloons and rathskellers.
The laws of city and state affecting social questions.
Our prison system—what help have the men?
Our various relief agencies—how far do they co-operate?”
ONE OF DAYTON’S MENACES
A heap of dead horses awaiting skinning and rendering at the

fertilizer plant
HEALTH
SANITATION AT DAYTON
[The widespread flood disaster in Ohio during the last week of
March led members of the Pittsburgh Flood Commission to study
the situation. Morris Knowles, a member of the Engineering
Committee of this commission, has had two assistants in the field
for this purpose. One of these, M. R. Scharff, who had previously
been employed by Mr. Knowles in making a sanitary survey of
the coal-mining camps in Alabama, paid particular attention to
the sanitary conditions resulting from the flood. The present
article embodies observations made on this trip.—Ed.]
Following in the wake of great disasters which descend from time
to time upon our cities, paralyzing the public services that make
crowded city conditions possible, is the outcropping of disease that
may, if unchecked, prove more disastrous even than the catastrophe
itself. This tendency was discernible in the first reports of the floods
that have recently devastated Ohio, Indiana and adjoining states, due
to the heavy rains of March 24–28. Nearly every flooded city
reported that its water works plant had been put out of commission,
or the water supply polluted, which with the increased chance of
infection, and the general lowering of vitality presented a situation of
unusual menace and one demanding complete and immediate
handling.
The most serious situation is Dayton, for here every sanitary
problem presented at any other point was involved. The complete,
immediate and effective organization to handle the situation which
was formed there was typical of the effective work now done at such
emergency periods.
At Dayton the water works plant was incapacitated by water that
reached ten feet above the boiler grates; there was unknown damage
to water distribution and sanitary sewerage and drainage systems;
storm sewers and catch basins were clogged with filth and debris;
dead animals were strewn on every side; the population was at high
nervous tension, their vitality lowered by shock, exposure, cold, and
lack of food and drink; hundreds of people were crowded for days in
single buildings or dwellings; thousands, probably, had been exposed
to intestinal infection by drinking the dirty flood water as it swirled
through the streets; hundreds had only wet cellars and rooms to
return to, if their homes were not altogether destroyed; and
everywhere on everything—walls, ceilings, floors, furniture, streets
and sidewalks—was a thick coating of the black, sticky, slimy mud
left by the retreating waters. This in a measure pictures the situation
at Dayton as the flood waters receded. And Dayton knew at once that
the toll of the flood would be as nothing compared to the pestilence,
unless attention and energy were directed to these problems.
This appreciation of the paramount importance of sanitation was a
striking revelation of the success of the campaign of sanitary
education that has characterized the last century. In every phase of
the work of recovery, in the warning signs and directions on almost
every post, in the placards on the automobiles of the sanitary
department stating that “This car must not be stopped or delayed day
or night,” in the daily exhortations in the free newspapers distributed
throughout the city, in a thousand ways, Dayton declared again and
again:
“Sanitation first and foremost. Then everything else.”
Such was the spirit of the members of the Dayton Bicycle Club,
when they met as the waters receded from their club-house to
consider what service they could best render to their stricken city,
and volunteered to remove the dead animals strewn it the streets.
Such also was the message reiterated by the Ohio State Board of
Health, the city health officials, the representatives of the national
government, the Red Cross, the Relief Committee, the Ohio National
Guard, and every one of the splendid organizations that are working
shoulder to shoulder to clean up Dayton and to prevent conditions
more costly in toll of life than the deluge itself.
One of the remarkable features of the handling of the relief work at
Dayton was the entire absence of red tape, the lack of conflict, and
the universal evidence of harmonious co-operation between the
various organizations at work, notwithstanding that there was no
complete centralization of direction and that some of the
organizations were proceeding practically independent of the others.
“Results, not credit,” was the watchword, and the results were such
as to reflect the most lasting credit upon all engaged in the work.
The Dayton Bicycle Club showed wisdom in volunteering to
remove the dead animals from the street. Nearly every horse in the
more than seven square miles of the city that was under water—and
this area contained all the important livery stables—was drowned,
and quick action was needed to remove the bodies to prevent serious
results. A sanitary department was organized, and as rapidly as
automobile trucks and wagons were volunteered, they were pressed
into service. Over 100 vehicles and about 600 men were engaged on
this work. A rendering company, which handles all the garbage
collected in the city, agreed to take care of the horses and did so as
fast as they came for a time. When the carcasses came so rapidly that
it was necessary to heap them up on the grounds of the plant, and
then on a vacant field nearby, the plant was a grewsome place
indeed. Up to the night of March 31, 1,002 had been received. A
number were picked up the next two days, so that the final total was
probably in the neighborhood of 1,100.
At about the time this work was started, a reconstruction
department was organized, under the Citizens Relief Committee,
with divisions, each under an engineer, assigned to street cleaning,
sewers and drains, streets, and levees. By March 31, the removal of
dead animals had been practically completed, and the organization
and equipment of the sanitary department were merged with those
of the street cleaning division of the reconstruction department.
Sanitary notices directed that all mud and rubbish be deposited at
the curb, the city was divided into districts and collection progressed
rapidly, considering the wagons and trucks available. More wagons
could have been put into service, but horses were lacking. All mud
and rubbish was hauled to one of the half-dozen city rubbish dumps
located in low outlying sections, or was dumped off bridges into the
river. The employes of the city water works department were able to
get into the pumping station on March 28 and the following day
pumping was resumed. Dayton’s water supply comes from a number
of deep drilled wells along the Mad River. It is pumped direct into
the mains without storage, by means of a Holly vertical, triple-
expansion, crank and fly-wheel engine. This pump has given rise to
the local name of “Hollywater” applied to the city supply. It was
feared at first that the distribution system had been badly damaged,
but investigation showed that only three small mains had been
broken. Water, at reduced pressure, was therefore possible, except in
one or two small sections.
AN IMPROVISED COMFORT
STATION
Dayton water is exceptionally pure, but it was feared that there

might have been leakage of flood water into the pipes while the
pressure was cut off and so notices to “boil all water, even the
Hollywater” were posted. Samples were promptly taken for analysis
from various portions of the distribution system by the chemist of
the National Cash Register Company, the bacteriologist of the city
Board of Health, and by the State Board of Health, but the injunction
to boil water was continued, even though the first analysis was
favorable.
The catch basins and storm sewers throughout the city were badly
clogged with wreckage and filth, and early cleaning was imperative.
The city was divided into seven drainage districts, and gangs of men
and wagons assigned to shoveling out catch basins and hauling the
rubbish to the dumps. At the same time systematic inspection of the
sanitary sewerage system was begun. It had been expected that the
sewers would be clogged, like the storm drains, and the early sanitary
notices issued contained these warnings:
IMPORTANT
Sanitary Notice
FOR YOUR OWN HEALTH
(1.) Do not use Sanitary sewers and Closets until notified by the Board of
Health. Even if the hollywater system is on, the sewers are full of mud and
will clog. Burn or bury all excreta garbage and filth. Add lime and bury deep.
Use disinfectant in out-door trenches also.
(2.) Thoroughly scrub, clean and dry your cellar. Keep your cellar
windows open. Remove and burn or bury all rubbish. Sprinkle lime around
cellar, especially in damp places. Sprinkle floor with disinfectant sent
herewith (two tablespoons-full to one quart of water.)
(3.) Thoroughly clean your in and out door premises.
(4.) Place concentrated lye or a tablespoon of disinfectant in each sink or
trap in toilet, basement and kitchen. Allow to stand over night. Do this every
evening.
(5.) Boil all water, even holly water, and thoroughly cook all food. Boil all
cooking utensils. Do this for months to come.
(6.) Do not enter houses which have been flooded until thoroughly
cleaned and dried.
(7.) Keep your own self clean.
Do these things to avoid pestilence and sickness.
Do it for yourself.
Do it for Dayton.
Take care of yourself and you will take care of Dayton.
Maj. L. T. Rhoades,
U. S. Army.
ONE OF THE EARLY NOTICES
“Do not use water closets. Contents will reach cellars. Use vessels, disinfect, and
bury in back-yards. Disinfectants: carbolic acid, chloride of lime, bichloride of
mercury, and creolin.”
“Do not use sanitary sewers and closets until notified by the Board of Health.
Even if the “Hollywater” system is on, the sewers are full of mud and will clog.
Burn or bury all excreta, garbage and filth. Add lime and bury deep. Use
disinfectant in out-door trenches also.”
Inspection showed a much better condition than was anticipated.
In all but three districts, the sanitary sewers were running freely and
the warnings were replaced by new notices:
“Sewers are open and ready for use. If the water supply is not sufficient for
flushing, fill the tank of the closet with a bucketful of water, and flush as usual.”
Wooden public convenience stations were also established over
sewer manholes in the business sections and in residential sections
without sewer connections.
The three sewer districts that were out of commission were the St.
Francis, the North Dayton, and the Riverdale low line. The St.
Francis sewer is a gravity line, and a manhole at the lower end was
completely choked up. It was necessary finally to dynamite this
manhole in order to open the line. The two latter lines are both low,
and sewage has to be pumped into the river by pneumatic ejectors.
The air lines from the compressor plant in the water works pumping
station were laid in the levees which were washed out and at one
point about 200 feet of pipe was lost. This was difficult to repair, and
these districts had to be left without sewerage until April 2, when a
by-pass on each line into the storm drains was opened, and the
backed-up sewage lowered sufficiently to clear most of the cellars
and to permit the use of water closets.
While this work was proceeding the organizations devoting their
energies to control of infectious disease, inspection, and
administration had been far from idle. The State Board of Health had
three sanitary engineers and two physicians, trained in public health
work, in the city before the waters receded. The city Board of Health
was one of the first in the field, and the medical corps of the Ohio
National Guard promptly took up the work. Co-operating with one
another, under the direction of Major L. T. Rhoades of the United

Ebook Advances in Food and Nutrition Research PDF Full Chapter PDF

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Ebook Advances in Food and Nutrition Research PDF Full Chapter PDF

Uploaded by

Copyright:

Available Formats

Advances in Food and Nutrition

Research - eBook PDF

First edition 2022

Copyright © 2022 Elsevier Inc. All rights reserved.

For information on all Academic Press publications

Publisher: Zoe Kruze

and quantitation, and in silico approaches. An analysis of posttranslational

constitutes an attractive alternative food preservation technique. Furthermore,

Peptidomics as a useful tool

2. Peptidomics in the characterization of peptides

Color, texture, flavour,

Bioactivities: antioxidant, antihypertensive,

Post-traductional Oxidation, deamidation,

Fig. 2 Major areas of peptidomics applications in foods.

and quantitation of bioactive peptides is shown in Fig. 3. The pretreatment

method of choice. Size-exclusion chromatography is based on the separation

2.1 Identification of peptides

bioactivities studied according to BIOPEP database (Minkiewicz et al.,

2.1.1 Identification of di-, tri-, and tetrapeptides

Dipeptide sequences can be repeated many times in protein sequences so

2.1.2 Pentapeptides or longer peptides identification

adjustment of the basic techniques in order to identify a larger number of

Fig. 8 Reversed-phase chromatographic separation after in vitro gastrointestinal diges-

conduct MSn fragmentation of the parent peptides, quadrupole/time-of-

spectrometry analysis revealed changes in the peptide profiles and evidenced

2.2 Quantification of peptides

provide a simple, reliable, versatile, and cost-effective quantification that can

optimization of the MRM parameters of low abundant peptides or small

after the quantification of the antihypertensive dipeptide AA at different

Protein of origin Food matrix

Bioactivity test in vitro and in vivo

Despite this procedure is very effective, it results very expensive and

where a is the number of fragments with given activity in a protein sequence,

The potential biological activity of protein (B) [μM1] is calculated with

where ai is the number of repetitions of i-th bioactive fragment in protein

4. Mechanisms of enzymatic hydrolysis of food proteins

Fig. 15 Scheme of food protein hydrolysis and enzymes involved. The

Proteins in foods Isolated food proteins

Endogenous peptidases Microbial peptidases Peptidases Microbial peptidases

Ripening Fermentation Reactor Fermenter

Small amounts of bioactive Large amounts of bioactive

Absorption in the organism

border intestinal epithelium proteases and blood stream enzymes that

5. Release of bioactive peptides in foods by endogenous

hydrolyze myofibrillar and sarcoplasmic proteins into large protein

The presence of dipeptides have been reported in fermented foods as a

6. Release of bioactive peptides through food proteins

content, temperature, redox potential) during the process. This usually

DETROIT BOOSTING FOR SAFETY

TRADE SCHOOL FOR PRINTERS

In Printing Trade News the recently established School for

A bill marking the initial step towards the establishment of state

MUNICIPAL MINIMUM WAGE

A minimum wage of 25s. ($6.08) a week for all able-bodied men

FULL CREW BILLS

An unusual publicity campaign on the part of railroads has

Edited by GRAHAM TAYLOR

TERRE HAUTE’S LABOR PARLIAMENT

George W. Greenleaf, secretary-treasurer of District Lodge No. 72,

Terre Haute’s Industrial Creed

The Presbyterian church in Canada does social service work

The changing relation of the synagogue and the community is

A PRAYER FOR EFFICIENCY