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Probiotic potential of noni juice fermented with lactic acid bacteria and
bifidobacteria
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Probiotic potential of noni juice fermented with lactic acid bacteria and
bifidobacteria
Chung-Yi Wang a; Chang-Chai Ng a; Hsuan Su a; Wen-Sheng Tzeng a; Yuan-Tay Shyu a
a
Department of Horticulture, National Taiwan University, Taipei, Taiwan
To cite this Article Wang, Chung-Yi, Ng, Chang-Chai, Su, Hsuan, Tzeng, Wen-Sheng and Shyu, Yuan-Tay(2009)'Probiotic potential of
noni juice fermented with lactic acid bacteria and bifidobacteria',International Journal of Food Sciences and Nutrition,
To link to this Article: DOI: 10.1080/09637480902755095
URL: http://dx.doi.org/10.1080/09637480902755095
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International Journal of Food Sciences and Nutrition
2009, 19, iFirst article
Abstract
The present study assesses the feasibility of noni as a raw substrate for the production of
probiotic noni juice by lactic acid bacteria (Lactobacillus casei and Lactobacillus plantarum) and
bifidobacteria (Bifidobacterium longum). Changes in pH, acidity, sugar content, cell survival and
antioxidant properties during fermentation were monitored. All tested strains grew well on noni
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juice, reaching nearly 109 colony-forming units/ml after 48 h fermentation. L. casei produced
less lactic acid than B. longum and L. plantarum. After 4 weeks of cold storage at 48C, B. longum
and L. plantarum survived under low-pH conditions in fermented noni juice. In contrast,
L. casei exhibited no cell viability after 3 weeks. Moreover, noni juice fermented with B. longum
had a high antioxidant capacity that did not differ significantly (P B0.05) from that of lactic acid
bacteria. Finally, we found that B. longum and L. plantarum are optimal probiotics for
fermentation with noni juice.
Introduction
The use of probiotic microorganisms to ferment food is traditional. Fermented
products may be part of a daily diet, improving the health and quality of life of
consumers. Most of the probiotic bacteria used in commercial products today are
members of the genera Lactobacillus and Bifidobacterium (Daly and Davis 1998). The
beneficial effects of probiotic bacteria in food include prophylaxis against some
intestinal infections, improvement of lactose digestion, control of gastrointestinal
infections, reduction of serum cholesterol levels and stimulation of the immune
system (Gilliland 1990; Lin et al. 1998; Salminen et al. 1998; Mattila-Sandholm et al.
1999). During the past few decades, fermented milk products such as yogurt have
become the most familiar probiotic products, and other foods fermented using
probiotic bacteria have been demonstrated to provide potential health benefits (Ou
et al. 2006). Recently, numerous lactic acid bacteria such as Lactobacillus acidophilius,
Lactobacillus plantarum, Lactobacillus casei, Bifidobacterium longum and Bifidobacterium
lactis have been used in fruits to produce probiotic beverage (Gardner et al. 2001;
Tien et al. 2005; Yoon et al. 2006; Kun et al. 2008). Chen et al. (2008) utilized
Correspondence: Prof. Yuan-Tay Shyu, Department of Horticulture, National Taiwan University, 140
Keelung Road Section 4, Taipei 10600, Taiwan. Tel: 886 2 33664850. E-mail: [email protected]
various ginger plants as the fermentation substrate. Ginger juice fermented with three
probiotic lactic acid bacteria has a stronger antioxidant capability.
Noni (Morinda citrifolia) is a tropical and subtropical plant that grows on the Pacific
islands. It has been of special interest to Polynesians for over 2,000 years because of its
natural medicinal qualities (Dixon et al. 1999; McClatchey 2002). Different parts of
the plant, including the fruits, leaves, bark and root, have been established to contain
various biologically active compounds (Chan-Blanco et al. 2006). In traditional
pharmacopoeia, the juice, extracts or isolated biological compounds from the noni
are claimed to prevent and cure numerous diseases. There are adopted primarily
to stimulate the immune system, to inhibit low-density-lipoprotein oxidation, to
scavenge free radicals, to provide anti-inflammatory benefits and to regulate of
cholesterols (Chong et al. 2004; Kamiya et al. 2004; Su et al. 2005; Basu and Hazra
2006; Calzuola et al. 2006). Most noni fruit is consumed as juice, which is
traditionally made by the natural fermentation of noni fruit in sealed containers for
48 weeks at ambient temperature (Wang et al. 2008). During the fermentation
process, the temperature, oxygen and microorganisms can cause undesirable chemical
reactions, which normally reduce the health benefits of the noni juice (Chan-Blanco
et al. 2007).
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The present study demonstrates the fermenting of noni fruit with probiotic lactic
acid bacteria and bifidobacteria strains to select an appropriate starter culture for
developing a probiotic noni juice. Accordingly, chemical and microbiological changes,
the number of viable cells and antioxidant properties during fermentation were
monitored.
for 72 h. Samples were taken at 0, 24, 48 and 72 h for chemical and microbiological
analysis.
Reducing power
The reducing power was measured using the approach of Duh and Yen (1997). One
milliliter of NJS, phosphate buffer (0.2 M, pH 6.6, 0.5 ml; Merck), and potassium
hexacyanoferrate solution (1% v/w, 2.5 ml; Merck) were mixed and heated at 508C for
20 min. After the mixture had been cooled to room temperature, 0.5 ml of 10%
trichloroacetic acid (Merck) was added. Following centrifugation at 3,000 g for
10 min, an 1 ml aliquot of the supernatant was mixed with 1 ml distilled water and
0.1 ml ferric chloride (0.1%; Merck) and the reaction was then maintained for 10 min.
4 C.-Y. Wang et al.
Statistical analysis
All experimental results were mean of triplicate. The data were recorded as the
mean9standard deviation (SD) and analysis was performed with a statistical analysis
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system (SAS Inc., Cary, NC, USA). One-way analysis was conducted using analysis of
variance. Significant differences between means were determined by Duncan’s
multiple range tests. Results were regarded as statistically significant at P B0.05.
Each value represents the mean9SD (n3). Data bearing different uppercase superscript letters in the
same column are significantly different (PB0.05).
Probiotic potential of fermented noni juice 5
Table II. Time course of lactic fermentation of noni juice by B. longum.
Each value represents the mean9SD (n3). Data bearing different uppercase superscript letters in the
same column are significantly different (PB0.05).
titratable acidity expressed as lactic acid. The fermentation substrate seemed to affect
the viability of the cultures during storage (Gardner et al. 2001), perhaps because
L. casei may require essential growth nutrients that may absent from noni juice.
In these assays, the number of viable cells of lactic acid bacteria and bifidobacter
reached almost 109 CFU/ml in the first 48 h in the noni juice, but the population
tended to decreased to 103 CFU/ml during 3 weeks of storage at 48C (Table IV).
A similar phenomenon has been reported in other studies (Gardner et al. 2001; Yoon
et al. 2006). L. plantarum was capable of surviving in the fermented noni juice at 48C
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for several weeks. The viable cell counts of L. plantarum and B. longum remained at
2.690.7 105 and 3.490.4 105, respectively, after 4 weeks of storage at 48C.
However, L. casei was unable to survive at low pH or high acidity conditions in
fermented noni juice at 48C, and was completely unviable after 3 weeks. To be
effective as a health promoter, a food product should contain 106 CFU/ml probiotics
(Angelov et al. 2005). Accordingly, the viability of the lactic acid bacteria is the most
important factor during refrigeration or frozen storage. The viability of probiotic
organisms depends on the oxygen level in the products, the oxygen permeability of the
package, the fermentation time, and the storage temperature (Shah 2000). The
viability of probiotic bacteria also depends on inhibitory substances such as lactic acid
that is produced during cold storage. Other factors that govern the loss of viability of
probiotic organisms are been the decrease in pH of the medium and accumulation of
organic acid as a result of growth and fermentation (Hood and Zottola 1988; Shah
and Jelen 1990). In the present study, both L. plantarum and B. longum survived at low
pH in the fermented noni juice
Each value represents the mean9SD (n3). Data bearing different uppercase superscript letters in the
same column are significantly different (PB0.05).
6 C.-Y. Wang et al.
Table IV. Effect of cold storage on the viability of lactic acid bacteria in fermented noni juice.
CFU/ml
antioxidative ability of intact cells and the intracellular cell-free extract of lactic acid
bacteria were determined using various antioxidant assays. Table V presents the
antioxidant activity of noni juice using different cultures of L. casei, B. longum and
L. plantarum. Notably, regardless of the starter used, the fermented noni juice
exhibited a high antioxidative ability, measured in terms of total antioxidant activity,
reducing power and DPPH radical scavenging activity. However, after 72 h of
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fermentation, the fermented noni juice that contained B. longum had a high
antioxidant activity of 77% that did not differ significantly (P B0.05) from that of
fermented noni juice that contained L. casei and L. plantarum (72% and 71%,
respectively) after 72 h of fermentation. Fermentation with B. longum reduced the
antioxidant activity below that at the beginning of experiment (77.6% versus 71.7%),
representing a decrease of approximately 8% over 4 weeks. A more apparent decrease
in the antioxidant activity was observed in the noni juice with L. casei (decreased about
12%). The reducing power in the noni juice fermented with lactic acid bacteria and
bifidobacteria did not change significantly during storage at 48C. Samples that were
fermented with B. longum exhibited a higher reducing power (0.42) than those of
fermented L. plantarum and L. casei, whose reducing powers were 0.38 and 0.31,
respectively. Similar results were obtained for DPPH radical scavenging activity: noni
juice fermented with B. longum retained the highest scavenging activity (70%). Four
weeks of storage at 48C reduced the DPPH radical scavenging activity of noni juice,
especially of that fermented with L. casei. Wang et al. (2008) noted the antioxidative
effects produced during the fermentation of various carbohydrates by common
intestinal lactic acid bacteria. The data in Table V reveal that the sugar content of noni
juice is probably a sufficient source of essential carbon for B. longum. Therefore,
depending on the strain used, bacterial metabolism may affect the total antioxidant
activity in noni juice.
Recent studies have reported the effect of fermentation on antioxidant properties in
some food products. Nazzaro et al. (2008) observed an increase in antioxidant
properties in carrot juice fermented upon fermentation by lactic acid bacteria. Tien
et al. (2005) adopted sugar apple (Annona squamosa L.) as a substrate for fermentation
with Lactobacillus delbrueckii, Lactobacillus paracasei and L. casei, and found no
significant different between fermented juice and fresh juice. The starter used in
this study also was similar to those used by Chen (2008), who also found B. longum to
be the optimal fermentation starter in ginger juice. The fermented sugar apple juice
exhibited antioxidant activity from 65% to 75% and DPPH scavenging efficiency as
high as 72%. In this work, fermented noni juice exhibited antioxidant activity,
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Total antioxidant capacity (%) Reducing power (absorbance 700 nm) DPPH free radical scavenging activity (%)
Time (weeks) L. plantarum B. longum L. casei L. plantarum B. longum L. casei L. plantarum B. longum L. casei
Reported values are the means9SD (n3). Data bearing uppercase superscript letters in the same column (different sampling time) and lowercase superscript letters in
the same row (different inoculation of starter) are significantly different (PB0.05).
7
8 C.-Y. Wang et al.
reducing activity and the scavenging effect of DPPH radicals, all of which varied with
the starters used, but L. casei and L. plantarum did not influence the antioxidant
activities of noni juice upon fermentation when compared with unfermented noni
juice (P B0.05). Noni juice that had been fermented with B. longum had greater
antioxidant activity than unfermented noni juice (Table V). Therefore, the increase in
the antioxidant activities varied with the starter organism.
Conclusion
The ability of lactic acid bacteria (L. casei and L. plantarum) and bifidobacteria
(B. longum) to utilize noni juice in cell synthesis and lactic acid production without
external nutrient supplement was investigated. These lactic cultures grew well in noni
juice at 308C, and the viable cell count was nearly 109 CFU/ml after 48 h of
fermentation. Both B. longum and L. plantarum retained viability at low pH and under
highly acidic conditions in fermented noni juice during cold storage at 48C. In
contrast, L. casei could not survive the low pH of fermented noni juice, and lost all
viability during 3 weeks of cold storage at 48C. The increase in antioxidant capability
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associated with B. longum driven fermentation did not differ significantly (P B0.05)
from that with L. casei and L. plantarum during 72 h of fermentation. Based on the
findings in this study, B. longum and L. plantarum are the optimal probiotics for the
production of a health beverage.
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