ARTICLE IN PRESS

International Dairy Journal 18 (2008) 685– 694

Contents lists available at ScienceDirect

International Dairy Journal

journal homepage: www.elsevier.com/locate/idairyj

Review

Lactose: Crystallization, hydrolysis and value-added derivatives

Michael G. Gänzle , Gottfried Haase, Paul Jelen
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alta., Canada

abstract

Lactose, the most abundant component of milk of most mammals, has been thoroughly studied for its
physico-chemical properties, crystallization behavior and importance as a fermentation medium.
Studies of various approaches to lactose modifications to increase its value as a food ingredient or
nutraceutical component are more recent and presently predominate the research interest concerning
lactose. This review, while summarizing briefly some physico-chemical properties and older studies
concerning crystallization behavior (mutarotatory equilibrium, solubility, crystalline habit and form)
focuses also on the modification alternatives to increase the utilization of lactose through value-added
products. Various approaches to lactose hydrolysis leading to increased solubility, higher sweetness and
expanded availability of milk and dairy products for lactose intolerant consumers are compared with an
emphasis on crude enzyme extracts. Principles and processes for conversion of lactose to lactitol,
lactobionic acid, lactulose, lactosucrose, and galacto-oligosaccharides are highlighted.
& 2008 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
2. Characteristics of lactose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
2.1. Occurrence and properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
2.2. Mutarotation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
2.3. Solubility and sweetness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
3. Lactose crystallization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
3.1. Crystalline habits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 686
3.2. Shapes and crystallization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687
3.3. Effects of acidity, alkalinity, and salts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687
3.4. Other milk components, chemicals and food ingredients . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687
3.5. Industrial process conditions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 687
3.6. Amorphous noncrystalline lactose (lactose glass) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 688
3.7. Lactose in food products—quality and industrial applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 688
4. Lactose hydrolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 688
4.1. Principles and theoretical considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 688
4.2. Industrially applicable alternatives for lactose hydrolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 688
4.3. Lactose hydrolysis by mechanically disrupted bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 689
4.4. Future prospects for industrial applications of lactose hydrolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 689
5. Conversion of lactose to value added materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 689
5.1. Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 689
5.2. Lactulose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 690
5.3. Lactitol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 690
5.4. Lactobionic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691
5.5. Lactosucrose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691
5.6. Galacto-oligosaccharides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691
6. Conclusions and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 692
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 693

 Corresponding author. Tel.: +1780 492 0774; fax: +1780 492 4265.
E-mail address: [email protected] (M.G. Gänzle).

0958-6946/$ - see front matter & 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.idairyj.2008.03.003
 ARTICLE IN PRESS

686 M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694

1. Introduction water is ½a20 

D ¼ þ89:4 (anhydrous weight basis). The melting
point is 201.6 1C. The most predominant crystals forms are prisms,
Prior to the seventeenth century, milk was considered to have pyramids, or ‘‘tomahawks’’ depending on the conditions of
only three components, curd, fat, and whey (Whittier, 1944). crystallization. The crystals are hard and not very soluble. If their
Bartolettus (1633) isolated an ‘‘essential salt without nitrogen’’ size in food products is over 10–16 mm they can be detected
from whey in 1633; Ettmueller (1688) isolated lactose from sensorically and create a defect called ‘‘sandiness’’.
evaporated whey and purified it by recrystallization (both authors The anomer of a-lactose is b-lactose. Since no water is
as cited by Whittier, 1944). associated with the molecule in this case, its designation is b-
During the 18th century, lactose became a commercial anhydride. It crystallizes from supersaturated lactose solutions at
commodity. The foundation of the present knowledge of lact- temperatures above 93.5 1C. Its specific optical rotation is ½a20D ¼
ose—especially regarding its chemistry and molecular struc- þ35:0 and it has a melting point of 252.2 1C. The b-anhydride
ture—was laid during the early 20th century. From this basis crystals, which are sweeter and considerably more soluble than
over the years, the present concept of understanding of the the a-hydrate, commonly occur as uneven sided diamonds when
characteristics and the utility of this unique sugar was developed. crystallized from water, and curved and needle-like prisms from
The objective of this somewhat historical but mainly forward- alcohol-based solutions.
looking review is to highlight some of the well-known properties
related to the roles of lactose in modern dairy and food industries, 2.2. Mutarotation
bearing on the ongoing quest to find solutions that utilize one of
the main dairy by-products—the cheese whey, of which lactose is In aqueous solutions, a- and b-lactose are present in equili-
the main component. brium. Regardless of the form used in preparing a solution, the
optical rotation will change the a form into the b form and vice-
versa through the process of mutarotation until ½a20 
D ¼ þ55:3 at
2. Characteristics of lactose the equilibrium (anhydrous weight basis). This is equivalent to
37.3% a-lactose and 62.7% b-lactose; this equilibrium ratio is
2.1. Occurrence and properties affected slightly by differences in temperature (Nickerson, 1962),
but not by differences in pH.
Lactose is present in the milk of all mammals with only a few The rate of mutarotation is greatly influenced by both
minor exceptions. The approximate concentration in mammalian temperature and pH as well as by other sugars and salts (Haase
milk is between 2.0% and 10% (Holsinger, 1988; Whittier, 1944). & Nickerson, 1966; Patel & Nickerson, 1970). The rate is slow at
The lactose content of bovine milk ranges between 4.4% and 5.2% low temperatures but increases 2.8 times with every 10 1C rise in
averaging at 4.8% anhydrous lactose. This compares with 7% in temperature, becoming almost instantaneous at about 75 1C. The
human milk. rate of mutarotation is at a minimum at about pH 5.0 increasing
Lactose (4-0-b-galactopyranosyl-D-glucopyranose, C12H22O11) with changes on either side of this value.
is a disaccharide comprising one glucose molecule linked to a
galactose molecule. A distinctive feature of lactose is its 2.3. Solubility and sweetness
manifestation in different states and temperature-dependent
physico-chemical interrelationships. Lactose in aqueous solutions The solubility of lactose is low compared with other disacchar-
is present in a and b forms (Fig. 1). Crystals of a-lactose can be ides but the effect of temperature on solubility is more pronounced
prepared as monohydrate by concentrating an aqueous lactose (Sienkiewicz & Riedel, 1990). The solubility of lactose is only about
solution to supersaturation and allowing it to crystallize at a 10% of that of sucrose at ambient temperature (Ryder, 1988). The a-
moderate rate below 93.5 1C (Drapier-Beche, Fanni, & Parmentier, and b-lactose have vastly different solubilities. The b-lactose is
1999; Gillis, 1920; Hudson, 1908). Its specific optical rotation in much more soluble in the ambient conditions and the a-lactose
above 93.5 1C. The mutarotatory equilibrium favoring the a variant
at ambient temperature results in the low overall solubility of
CH2OH CH2OH
lactose in these conditions and much higher solubility at
H temperatures close to 93 1C. The sweetness of lactose solutions at
C CH HC O ambient conditions is about 20% that of sucrose. However, the
H sweetness of lactose in milk is clearly noticeable and a sweet taste
H C OH C HC OH CH following addition of 0.75% lactose to regular skim milk could be
H H detected by trained taste panels (Jelen & Michel, 1999).
O C C OH
HO C O
H

OH OH 3. Lactose crystallization
α−lactose

Lactose can be present in dairy products in two crystalline

CH2OH CH2OH
forms, a-hydrate and b-anhydride, as well as an amorphous
H ‘‘glass’’ mixture of a- and b-forms. This non-crystalline lactose
C CH HC O OH glass contains the a- and b-forms in the same ratio as in the
H solution from which it was generated. Solutions of lactose are
H C OH C HC OH CH capable of being highly supersaturated before spontaneous
H H crystallization occurs.
O C C
HO C O
H
3.1. Crystalline habits
OH OH
β−lactose
Crystallization in its most basic concept is a two-step process
Fig. 1. Structure of lactose molecules in a and b configuration. involving nucleation and growth of the nucleus to a macro-size
 ARTICLE IN PRESS

M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694 687

(Nickerson, 1974). In general, the rate of crystal growth increases (Jelen & Coulter, 1973b; Smart, 1988). Some salts led to significantly
rapidly as supersaturation (or precipitation pressure, expressed as higher crystal growth rates, and their effect was concentration
ratio of actual concentration to solubility) is increased (Twieg & dependent. Thus, salt concentrations per se appear to be important
Nickerson, 1968; van Kreveld & Michaels, 1965). The rate is factors influencing growth rates, with each salt exerting a different
different for the different faces of the crystals, an effect that alters effect. In addition, some salts seem to change the shape of crystals
the shape of the crystals during growth. Crystal surface integra- leading to elongations, needle-like shapes or triangular flake forms
tion is a critical factor in the formation of crystals, while the (Bhargava & Jelen, 1996; Jelen & Coulter, 1973b).
diffusion rate of a-lactose to the surface is not rate-limiting
(Thurlby, 1976). The rate of lactose crystal growth is not much
3.4. Other milk components, chemicals and food ingredients
different from that of sucrose, when considering that the growth
of lactose crystals occurs predominantly or solely on only one
Methanol and ethanol accelerate crystallization by as much as
face—the bottom—of the pyramidal crystal (Bhargava & Jelen
30–60% even at low (1%) concentrations (Nickerson, 1974; van
1996; Jelen & Coulter, 1973a, 1973b). Several older investigations,
Kreveld & Michaels, 1965). Alcohols reduce the solubility of lactose
especially in the Netherlands at the Cooperative Condensfabriek
and support spontaneous nucleation. The latter effect, accelerating
Friesland, focussed on the mechanisms and kinetics of the lactose
the formation of crystals, may support the step theory (step
crystal formation and growth in great detail (e.g. van Kreveld,
generation and rapid growth) in lactose crystallization (Michaels &
1969; van Kreveld & Michaels, 1965; Visser, 1983).
van Kreveld, 1966; Nickerson & Moore, 1974a, b).
In dairy products, the presence of ‘‘impurities’’ or chemicals
3.2. Shapes and crystallization used as food additives may inhibit or accelerate the growth of
lactose crystals, alter crystal form (Jelen & Coulter, 1973b) or have
The pyramid, tomahawk and prism shapes are the most no effect. In some instances, impurities may inhibit the formation
common forms but lactose crystals can be observed in a variety of the nuclei. The effects on crystal growth are a function of
of other shapes, depending on the conditions of crystallization. intricate physico-chemical interactions of available energies,
The principal factor governing the form of lactose crystals is the adsorption rate to the various crystal faces, steric hindrance
supersaturation of the solution (Herrington, 1934). High super- caused by proteins and polysaccharides, tendency to spontaneous
saturation forces rapid crystallization and only prisms form. As nucleation, and the concentration of substances in question
supersaturation decreases, the dominant crystal form changes to (Nasirpour, Scher, Lindner, & Desobry, 2006; Nickerson, 1974). In
diamond-shaped plates, then to pyramids and tomahawks, and the presence of impurities, lactose crystals tend to be irregularly
finally to fully developed crystal showing a multitude of faces shaped and clumped, instead of yielding the characteristic crystals
(Holsinger, 1988; Jelen & Coulter, 1973a; Nickerson, 1979; van obtained from simple lactose solutions (Nickerson, 1962).
Kreveld & Michaels, 1965). More recent studies demonstrated the Gelatine is an example of a crystallization inhibitor that
physico-chemical interrelationships of aqueous lactose solutions reduces the rate by 25–60%. However, gelatine cannot suppress
at different degrees of supersaturation, temperatures, different nucleation in highly supersaturated lactose solutions, which
stages of crystallization and in the presence or absence of explains its ineffectiveness in preventing sandiness in ice cream
different water-miscible organic solvents (Zeng, Martin, Marriott, (Nickerson, 1962). On the other hand, various marine and
& Pritchard, 2000b). Although the majority of lactose crystals vegetable gums are now widely used in ice-cream formulations,
were found to be either tomahawk-shaped or pyramidal, at higher because they inhibit the formation of lactose crystal nuclei.
concentrations elongated cuboidal crystals were observed, with Carrageenan-containing systems have been shown to inhibit
higher initial lactose concentrations resulting in more elongated crystallization (Kouassi, Jouppila, & Roos, 2002). Gels can provide
particles. a ‘‘protective environment’’, thus presenting a ‘‘control’’ mechan-
ism for the rate of crystallization as well as the uniformity of the
crystals (Zeng, Martin, Marriott, & Pritchard, 2000a).
3.3. Effects of acidity, alkalinity, and salts

The pH is an important factor in lactose crystallization 3.5. Industrial process conditions

(Nickerson & Moore, 1974b). The effect has been attributed to its
influence on the rate of mutarotation, as crystallization removes Lactose crystallization is particularly complicated in complex
only one anomer from the equilibrium, which may become dairy systems subjected to industrial process conditions, such as
depleted in crystallizing form if mutarotation is slow. However, spray-drying, freeze-drying, and various storage situations. In
Twieg and Nickerson (1968) and Nickerson and Moore (1974b) these cases, the distinct crystals start to coexist with amorphous
demonstrated that mutarotation becomes limiting only when lactose. During storage, the rate of crystallization in dry lactose-
crystallization occurs rapidly with a large surface area. Alkaline containing powders increases with increasing relative humidity
conditions speed up crystallization but also favor formation of (Miao & Roos, 2005), and is higher in spray-dried materials than in
lactose degradation products that may inhibit crystallization. freeze-dried materials. The crystallization rate was lower in all
However, the addition of calcium lactate or K2HPO4 (both of lactose/protein mixtures than in pure lactose (Haque & Roos,
which increase pH) resulted in opposite effects on crystallization 2004). Proteins have a profound effect on lactose crystallization
(Bhargava & Jelen, 1996), indicating additional effects of the (Kouassi et al., 2002; Miao & Roos, 2005). Lactose crystallized
mineral composition and the ionic strength. mainly as a-lactose monohydrate in spray-dried lactose/whey
Lactose solubility values in whey UF permeate solutions as protein isolate and lactose/gelatin mixtures. Anhydrous b-lactose
well as model systems seem to relate directly to growth rates of and a-lactose monohydrate crystals formed in freeze-dried
lactose crystals although the rates of crystal growth were lower in lactose/whey protein isolate and lactose/gelatin mixtures, while
permeate than in pure lactose solutions (Bhargava & Jelen, 1996). only a-lactose monohydrate crystallized in both spray-dried and
Presence of non-lactose impurities such as riboflavin or KCl could freeze-dried lactose/Na-caseinate mixtures (Miao & Roos, 2005).
be responsible for this effect (Jelen & Coulter, 1973b; Smart, 1988). In freeze-dried skim milk, lactose crystallized as an anhydrous
When salts were added specifically to pure aqueous solutions, mixture of a- and b-lactose in a molar ratio of 5:3 (Jouppila,
various salts had very different effects on crystal growth rates Kansikas, & Roos, 1997).
 ARTICLE IN PRESS

688 M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694

3.6. Amorphous noncrystalline lactose (lactose glass) Table 1

Technological alternatives for production of sweetening syrups based on lactose
hydrolysis
When a lactose solution is dried rapidly, its viscosity increases
so quickly that crystallization cannot take place and the dry Process Main characteristics
lactose forms an amorphous (noncrystalline) lactose ‘‘glass’’.
Lactose in milk powder (spray, roller, or freeze-dried) is Acid-catalysed hydrolysis pHo1.5
Direct acid addition Temperature 90 1C
noncrystalline and exists in the same equilibrium mixture of
Ion exchange resin Temperature 150 1C
a- and b-lactose as existed in the milk prior to drying (Nickerson, Immobilized enzyme technology Reactor containing suitably
1974; Zadow, 1984). The glass transition temperature of lactose is immobilized enzyme
influenced by its concentration, relative humidity, and the Membrane-based enzyme reactor Free enzyme hydrolysis with enzyme
presence of other milk components (Jouppila & Roos, 1994; Roos separation and reuse
Free (soluble) purified enzymes Single use of the enzyme added to
& Karel, 1992). Stickiness of dairy powders was shown to be each batch and not recovered
related directly to the glass transition temperature (Paterson,
Brooks, Bronlund, & Foster, 2005).
glucose and galactose enables its use as sweetening syrups in
ice cream and other dairy and non-dairy foods. In addition, lactose
3.7. Lactose in food products—quality and industrial applications
hydrolysis increases the availability of dairy nutrients worldwide
by alleviating lactose intolerance. Saccharomyces cerevisiae and the
The crystallization principles and the factors influencing the
majority of other yeast species are lactose-negative and the
growth of lactose crystals have been applied to dairy products to
lactose hydrolysis is a prerequisite for whey conversion to baker’s
understand the consequences of processing conditions of lactose-
yeast, animal feeds based on yeast biomass, ethanol or other
containing milk products on their quality. It is generally under-
value-added conversions by yeasts. Numerous technical reviews
stood that rapid crystallization produces small crystals (Fox &
(e.g. Geilman, 1993; Gekas & Lopez-Leiva, 1985; Harju, 1987a;
McSweeney, 1998; Nickerson, 1974). Innovative designs of equip-
Shukla, 1975) can be consulted for details of the various technical
ment and appropriate processing techniques (such as seeding a
approaches to accomplish lactose hydrolysis (Table 1). Thus, the
sufficient amount of small crystals) enable the formulation of high
subject will be discussed here only briefly.
quality products even under high-yield process conditions.
Generally, all that is needed to hydrolyze lactose is an enzyme
One of the most objectionable texture defects in dairy products
or a chemical process breaking the bond connecting the two
is sandiness that may occur particularly in ice-cream and the
monosaccharides. Young mammals and certain bacteria, yeasts,
Norwegian whey cheese mysost. Sandiness is caused by lactose
moulds, and plants exhibit b-galactosidase activity (Pritzwald-
crystals which are large enough to be detectable in the mouth but
Stegman, 1986). The enzyme has been characterized thoroughly
which do not dissolve readily, thus producing a rough or gritty
(Wallenfels & Weil, 1972), and its properties and potential
sensation. Lactose glass in spray-dried milk and whey powders is
applicability in the dairy industry described and reviewed (Smart,
stable if protected from humidity, but is very hygroscopic, causing
1993). Alternatively, a chemical route for hydrolysis of lactose was
stickiness and caking (Haque & Roos, 2005; Holsinger, 1988;
known since the beginning of the 20th century (Whittier, 1925),
Paterson et al., 2005).
requiring extremely acidic conditions (pHo1.5) and very high
In the pharmaceutical industry, lactose is commonly used as a
temperatures (up to 150 1C). As an alternative to the enzymatic
bulking agent in pharmaceutical formulations for both human and
lactose hydrolysis, this approach has attracted renewed interest in
veterinary medicinal products (EMEA, 2002; Miao & Roos, 2005).
Australia, the Netherlands, and New Zealand (e.g., de Boer &
Uniformity of crystals, surface smoothness as well as favorable
Robbertsen, 1981; Haggett, 1976; MacBean, Hall, & Willman,
shapes are of critical importance for medical applications. The source
1979), and was even patented (Block, 1952) but proved to be of
and grade of lactose can have a substantial effect on drug delivery.
limited industrial applicability.
For example, in a dry-powder-inhaler application increasing the
elongation ratio of the lactose crystals improved the medical profile 4.2. Industrially applicable alternatives for lactose hydrolysis
of the drug in question significantly. The higher dispersibility and
fine particle fraction of the lactose appeared to be responsible factors The move towards industrial applications of the lactose
(Larhrib, Martin, Prime, & Marriott, 2003). hydrolysis process gained momentum in mid-1980s. In the
International Dairy Federation, a very active Group of Experts
under the leadership of Wayne Modler was set up to foster
4. Lactose hydrolysis international cooperation on the subject, resulting in production
of a valuable technical monograph (IDF, 1993). Major projects in
4.1. Principles and theoretical considerations UK, Australia, Finland and several other countries were driven by
perceived major market opportunities. Unfortunately, most of
The explosive rise in cheese production in the second part of them proved to be deceptive as in the case of the Australian
the last century necessitated the search for new alternatives for initiative that focussed on the Far East (Zadow, 1993). The two
whey utilization, which means predominantly finding uses for major approaches included the ‘‘free enzyme’’ route and the
lactose. The uses for traditional crystalline lactose remained static, development of immobilized enzyme reactors. Several b-galacto-
or even declined as new, cheaper sources of fermentable sidase preparations were commercialized by large industrial
carbohydrate became available. The theoretically simple but producers in the 1990s, at the peak of these developments.
industrially and especially economically challenging process of The free enzyme route appeared initially to be the preferred
hydrolyzing lactose for production of sweetening syrups gained mode for production of lactose hydrolyzed milk and is still being
prominence in the lactose literature in the latter part of the 20th used today. Although the enzyme preparation cannot be reused,
century. The aims of these industrial developments were two- which represents a major economic disadvantage, ‘‘lactose-free’’
fold—to use lactose as a sweetening carbohydrate; and to enable milk produced by this method is available in limited quantities on
fermentation by lactose-negative microorganisms. Hydrolysis of retail shelves in many countries. Over-the-counter preparations of
lactose into the much sweeter monosaccharide components the soluble b-galactosidase such as Lactaids are sold for use by
 ARTICLE IN PRESS

M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694 689

lactose-intolerant consumers as pills, providing time-limited Table 2

relief of lactose intolerance after ingesting lactose-containing Current and potential industrial applications of the lactose hydrolysis process in
mainstream dairy processing or utilization of dairy by-products
foods. The ‘‘tetra-lacta’’ process by the Tetra-Pak company for
production of UHT lactose-free milk was based on dosing very  Lactose hydrolysis in fluid milk products
small quantities of sterile b-galactosidase preparation into  Control of lactose crystallization in concentrated dairy products (whey
product at the time of aseptic packaging-lactose hydrolysis being cheeses, ice cream, sweetened condensed milk)
accomplished during the unrefrigerated shelf life of the product.  Whey and permeate applications (food and non-food uses)
A very successful industrial application of the soluble enzyme  Formulated pet food applications
approach has been developed recently by the Finnish company  Production of oligosaccharides and exopolysaccharides

Valio, one of the principal industrial pioneers in the field of lactose  Enhancement of starter culture production

hydrolysis. The patented process is based on removing most of the  Development of new flavors

lactose from regular milk by a chromatographic process developed  New product ideas combining lactose hydrolysis and flavor developments

by Harju (1987b), followed by hydrolysis of the remaining lactose  New/modified products (whey cheeses, spreads, drinks, ‘‘milk honey’’, dairy-
based confectionery)
by a soluble enzyme. Now marketed successfully in several
Scandinavian and other European countries, the Finnish process
results in comparable taste to the regular cows milk and avoids
the uncharacteristic excess sweetness caused by liberated glucose cells and production of ‘‘enzyme cocktails’’ with high b-galacto-
and galactose associated with traditional in situ lactose hydrolysis sidase activities. The application of these ‘‘in-house’’ produced
methods (Jelen & Tossavainen, 2004). cocktails for lactose hydrolysis under carefully selected conditions
The requirement for enzyme preparations in the free enzyme could result in optimal hydrolysis rates, without significant
approach is reduced by membrane reactors in which the soluble detrimental effects of other enzymatic processes. Our research,
enzymes are being recovered for repeated use (e.g., Mehaia, using a high b-galactosidase producer Lactobacillus delbrueckii
Alvarez, & Cheryan, 1993). However, the practicality and econom- subsp. bulgaricus strain ATCC 11842, focussed on the economic
ical feasibility of this technology in industrial applications seems (Bury & Jelen, 2000) and technical (Kreft, Roth, & Jelen, 2001)
doubtful. Effectiveness of membrane reactors for lactose hydro- feasibility of the process; optimization of the growth media
lysis in milk or whey is affected by several of the typical problems (Vasiljevic & Jelen, 2001) and of the mechanical disruption process
of the membrane processes (membrane fouling, membrane (Bury, Jelen, & Kalab, 2001; Geciova, Giesova, & Jelen, 2002);
selectivity, effects of some solutes on flux) as well as by the suppression of competing enzymatic reactions (Vasiljevic & Jelen,
gradual loss of enzyme activity. 2002); stability of the enzyme during the hydrolysis process (Kreft &
At the height of the ‘‘lactose hydrolysis euphoria’’ in the mid- Jelen, 2000); and sensory aspects of the final products (Vasiljevic,
1980s, developments of the immobilized enzyme technology Wismer, & Jelen, 2003). Clearly, before this approach could become
appeared to offer the ultimate solution. The much publicized successful, additional research would be needed, including a search
‘‘Corning process’’, involving enzyme immobilization on sintered for a more effective microbial producer of the enzyme and/or
glass beads, seemed to be ‘‘winning the race’’ of the day involving genetic engineering leading to significant overproduction
(Anonymous, 1981, 1984). Unfortunately, even in this very of the b-galactosidase and suppression of some of the other enzymes
promising case, the unfavorable economic realities and technical catalyzing potentially undesirable reactions.
difficulties resulted its final demise. Several other commercial
processes using the immobilized enzyme approach were tabu-
4.4. Future prospects for industrial applications of lactose hydrolysis
lated by Harju (1987a) and the most prominent ones described
briefly by Sienkiewicz and Riedel (1990). The Finnish develop-
ment by the Valio company, based on the b-galactosidase Lactose hydrolysis as a major industrial process appears to be
immobilization on a special resin, is still being used for grossly underutilized today. The amount of lactose available just
production of a successful whey drink (Jelen, personal observa- in the dried whey produced in Europe and North America can be
tion, 2006) as well as for production of some of the many lactose- estimated as at least 1.4 MT annually (IDF, 2005). In addition to
hydrolyzed dairy products marketed under the HYLA (HYdro- one of the main uses of lactose hydrolysis for alleviation of the
lyzed-LActose) brand (Jelen & Tossavainen, 2004). lactose malabsorption problem, other prospective applications
both in mainstream processing and as a route for by-products
valorization are listed in Table 2. In this regard, lactose hydrolysis
4.3. Lactose hydrolysis by mechanically disrupted bacteria can be considered as belonging to the family of enzymatically
catalyzed or chemical processes leading to added-value conver-
The unfavorable economies of either free or immobilized sion of lactose to other industrially attractive products.
enzyme routes limit the large-scale applications of lactose
hydrolysis for many potential uses. In an attempt to find an
inexpensive solution to this problem, a recent research project has 5. Conversion of lactose to value added materials
been carried out within the framework of an NSERC-supported
Canadian Research Network centered at the Université Laval. The 5.1. Overview
project aimed at using traditional dairy bacteria, known to be high
producers of the b-galactosidase, as an ‘‘in-house’’ source of the An overview on commercially produced derivatives of lactose
enzyme that could be produced by the potential users themselves, is shown in Fig. 2 and their properties and applications are
similarly to the traditional production of bulk cheese starters in outlined below. Currently, only a small proportion of the lactose is
the cheese factories. However, in contrast to starter cultures for used as feedstock for the chemical, enzymatic or microbiological
use in cheese, the fermentative activities of b-galactosidase conversion to lactose derivatives. However, because the market
producing bacteria in the subsequent lactose hydrolysis process value is substantially higher than lactose, these compounds
would be detrimental to many such applications. Thus, the second provide significant opportunities for value-added conversion of
important aspect of this approach is the mechanical disruption of lactose to functional food ingredients. In recent years, attention
the bacterial cultures, resulting in the inactivation of the bacterial was focussed on prebiotic lactose derivatives. Prebiotics were
 ARTICLE IN PRESS

690 M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694

O OH
HO O OH
O O OH HO OH
HO
OH O O OH
HO
HO OH
OH Lactobionic acid HO OH
OH Lactulose

isomerisation
oxidation

OH O OH
HO transgalactosylation O OH
HO HO
O O OH O O OH OH
HO O O
HO OH O O
OH OH
HO
HO OH reduction HO OH
HO OH
OH Lactitol OH Lactose HO OH OH
OH

hydrolysis Galacto-oligosaccharides
fructosyl transfer (β-(1-4) or β− (1-6) linkage)

HO HO
HO
O O O
HO OH HO OH OH
O O
HO
OH HO OH HO OH
OH O O OH
HO
Galactose + Glucose OH
HO OH
OH Lactosucrose

Fig. 2. Overview on commercially produced lactose derivatives.

defined in 1995 as ‘‘non digestible food ingredients that to act as soluble receptors for pathogens (Kunz et al., 2000). While
beneficially affect the host by selectively stimulating the growth functions other than the prebiotic effects cannot be mimicked by
and/or activity of one or a limited number of bacteria in the colon, lactose derivatives, the addition of prebiotic oligosaccharides to
and thus improve host health’’ (Gibson & Roberfroid, 1995). infant formulae resulted in levels of bifidobacteria in the intestine
Prebiotics are frequently referred to as ‘‘bifidus factor’’ in the older that was comparable to the one found in breast-fed infants
literature. Prebiotic compounds are not digested and adsorbed in (Veereman-Wauters, 2005).
the small intestine, therefore, they are classified as dietary fibre
and contribute to a fibre-rich diet recognized as beneficial to
5.2. Lactulose
human health (Englyst & Englyst, 2005). The end products of the
metabolism of prebiotics by intestinal bacteria are lactate and
As small quantities of lactulose are formed during heating of
short chain fatty acids (SCFA) which contribute to the beneficial
milk, it has been used as an indicator for milk heat treatment
effects of prebiotics (Topping & Clifton, 2001). Moreover, pre-
(Elliott, Datta, Amenu, & Deeth, 2005). Lactulose is produced by
biotics are generally applied to stimulate growth and metabolism
isomerization of lactose in alkaline solution; boric acid shifts the
of bifidobacteria and the consumption of prebiotics results in an
isomerization equilibrium in favor of lactulose and prevents side
increase in both the occurrence and number of bifidobacteria
reactions. Lactulose, the first lactose derivative that was commer-
isolated from fecal material (Böhm et al., 2004; Cummings,
cialized (Kozempel, Kurantz, Craig, & Hicks, 1995; Timmermans,
Macfarlane, & Englyst, 2001). The establishment of metabolically
1997), can be produced enzymatically with b-galactosidases and
active bifidobacteria is considered to be beneficial for the host.
fructose as galactosyl acceptor (Lee, Kim, & Oh, 2004; Mayer et al.,
Beneficial effects may include the regulation of bowel habit,
2004). The relative sweetness of lactulose is 0.6 in comparison to
stabilization of the gut mucosal barrier and the prevention of
sucrose. Lactulose is a non-digestible carbohydrate which is
diarrhea, and increased mineral absorption (Cummings et al.,
fermented in the human colon (Nilsson & Nyman, 2005) but its
2001; Hopkins & Macfarlane, 2003; Lee, 1999). The application of
specific bifidogenic effects is less pronounced when compared with
prebiotics in infant foods is especially relevant to substitute the
fructo-oligosaccharides or galacto-oligosaccharides (Bouhnik et al.,
bifidogenic effect of oligosaccharides that are present in human
2004). Lactulose is widely applied as laxative and administration of
milk at a level of 1–1.3%, whereas the milk of ruminants has much
lactulose or lactitol is a standard treatment for chronic hepatic
lower concentrations of oligosaccharides (Kunz, Rudloff, Baier,
encephalopathy, although the scientific basis for the latter therapy
Klein, & Strobel, 2000). More than 130 different human milk
is questionable (Als-Nielsen, Gluud, & Gluud, 2004).
oligosaccharides were identified. Most compounds carry lactose at
the reducing end and are substituted with N-acetylglucosamine, L-
fucose, and/or N-acetylneuraminic acid (sialic acid). In addition to 5.3. Lactitol
the modulation of the microbial colonization of infants, possible
functions of these compounds are the stimulation of the immune Lactitol is produced by chemical hydrogenation of lactose; the
system, anti-microbial and/or anti-inflammatory protection, and relative sweetness of lactitol is 0.3–0.4 compared with sucrose.
 ARTICLE IN PRESS

M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694 691

Lactitol hydrolysis to galactose and sorbitol in the human hydrolyze lactose to glucose and galactose and alternatively
intestine is dependent on the intestinal microflora; most of the catalyze the transgalactosylation of lactose to produce galacto-
lactitol is metabolized to short chain fatty acids by the colonic oligosaccharides (Matthews, 2005, Fig. 3). Depending on the
microflora while a part of the galactose is resorbed (for review, see source of the enzymes, the oligosaccharides have predominantly
Dills, 1989). Lactitol is a strong laxative and its metabolism by b(1-4) and/or b(1-6) linkages. For example, the b-galactosidase of
humans is insulin-independent. Polyols such as mannitol, xylitol Kluyveromyces lactis produced predominantly b-(1-6) oligosac-
or arabitol, lactitol are applied as non-caloric sweeteners in charides (60 -galactosyl-lactose and b-D-Gal(1-6)D-Gluc), a b-
calorie-reduced and diabetic foods. Moreover, lactitol is used as an galactosidase of Sterigmatomyces elviae produced predominantly
alternative to lactulose in the treatment of hepatic encephalo- 40 -galactosyl-lactose whereas Bacillus circulans b-galactosidase
pathy (Als-Nielsen et al., 2004). forms b-(1-2), b-(1-3), b-(1-4) or b-(1-6) linkages to produce a
large variety of oligosaccharides (Asp, Burvall, Dahlqvist, Hallgren,
5.4. Lactobionic acid & Lundblad, 1980; Onishi, Yamashiro, & Yokozeki, 1995; Yanahira
et al., 1995). Glucose, galactose, mannose, fructose, maltodextrins,
Lactobionic acid is commercially produced by chemical oxida- N-acetylneuraminic acid, glucuronic acid and a number of
tion of lactose (Gerling, 1997). Alternative methods for the aromatic compounds have been shown to act as galactose-
production of lactobionic acid employing a glucose-fructose acceptor for b-galactosidases, providing a virtually unlimited
oxidoreductase from Zymomonas mobilis were described (Satory variety of oligosaccharides (Bridiau, Taboubi, Marzouki, Legoy, &
et al., 1997). Lactobionate is a strong chelator of calcium and is used Maugard, 2006; Lee et al., 2004; Miyasato & Ajisaka, 2004;
in calcium supplements in pharmaceuticals. The chelating proper- Takada, Ogawa, Saito, Murata, & Usui, 1998; Yanahira et al., 1998,
ties of lactobionate also enable applications as ion sequestrant in Fig. 3). Both 40 -galactosyl lactose and 60 -galactosyl lactose are
detergent solutions (Gerling, 1997). Moreover, lactobionate at a considered to have prebiotic properties (for review, see van Loo et
concentration of 100 mmol L1 is a key component of solutions al., 1999). However, different oligosaccharide preparations may
used for the cold storage of transplant organs. The cell impermeant vary with respect to technological benefits such as flavor
lactobionate prevents hypothermically induced cell swelling; in enhancing properties, sweetness, hygroscopicity, and solubility.
addition, iron chelation by lactobionate is thought to reduce The transgalactosylation reaction of b-galactosidases is favored
oxidative injury during storage (Southard & Belzer, 1995). at high lactose concentrations (Huber, Kurz, & Wallenfels, 1976)
and typical oligosaccharide yields range from 10% to 40% (GOS/
initial lactose). The effect of lactose concentration on lactose
5.5. Lactosucrose
turnover by a b-galactosidase from L. delbrueckii spp. bulgaricus is
depicted in Fig. 4. High temperatures enable increased initial
The trisaccharide lactosucrose, derived from transfructosylation lactose concentration (Boon, Janssen, & van’t Riet, 2000; Bruins,
of lactose, is commercially produced in Japan. Its sweetness relative Strubel, van Lieshout, Janssen, & Boom, 2003). Moreover, high
to sucrose is 0.3–0.6. Lactosucrose is not resorbed in the upper incubation temperatures strongly favored oligosaccharide forma-
intestine and is, thus, available for hydrolysis and metabolism by tion over lactose hydrolysis by b-galactosidase from L. delbrueckii
the colonic microflora. Lactosucrose has a bifidogenic effect and its spp. bulgaricus and the optimum yield of GOS was achieved under
consumption was reported to decrease fecal pH and to inhibit denaturing conditions at 50 1C (Fig. 4, Vasiljevic & Jelen, 2003).
growth of colonic clostridia (Ogata et al., 1993). However, the Other approaches to optimize oligosaccharide formation by b-
prebiotic effect of lactosucrose is not as well documented as is the galactosidases include screening for enzymes with high prefer-
case for fructo-oligosaccharides or galacto-oligosaccharides. ence for trangalactosylation (Boon et al., 2000; Cheng et al., 2006;
Transfructosylation of lactose is carried out by bacterial or fungal Vasiljevic & Jelen, 2003), enzyme immobilization to achieve an
fructosyltransferases using sucrose or raffinose as fructosyldonor. improved enzyme stability at high temperatures (Albayrak &
Fructosyltransferases catalyze the hydrolysis of sucrose as well as Yang, 2002), and use of high hydrostatic pressure to improve the
the transfructosylation to acceptor carbohydrates. Fructosyltrans- yield of oligosaccharides (Kawade, Sakakibara, Nomura, Suzuki, &
ferases catalyze the formation of high molecular weight fructan Kunugi, 1999). Moreover, protein engineering was successfully
polymers and the formation of oligosaccharides in addition to employed to obtain truncated or modified b-galactosidases with
sucrose hydrolysis. Depending on the type of polymer formed, high preference for production of oligosaccharides (Jørgensen,
fructosyltransferases are referred to as levansucrases or inulosu- Hansen, & Sougaard, 2001).
crases (Tieking & Gänzle, 2005; van Hijum, Kralj, Ozimek, The use of lactic acid bacteria (LAB) as producers of b-
Kijkhuizen, & van Geel Schutten, 2006). The formation of lactosu- galactosidase enzymes offers substantial potential for the produc-
crose by levansucrase was initially described using an enzyme from tion of GOS. First, LAB are known to be good producers of
Rhanella aquatilis (Hestrin & Avigad, 1958; Ohtsuka et al., 1992). extracellular b-galactosidases that enable GOS production from
Because the spectrum of acceptor-carbohydrates is essentially lactose (Garman, Coolbear, & Smart, 1996; Hung, Xia, Hu, & Lee,
comparable in all bacterial levansucrases characterized so far (Cote 2001; Smart, 1991). Second, LAB have a safe tradition in food
& Ahlgren, 1993; Hestrin & Avigad, 1958; Tieking, Kühnl, & Gänzle, fermentations and exhibit rapid anaerobic growth on agricultural
2005), it can be assumed that the synthesis of lactosucrose from substrates including waste products such as whey. Therefore, GOS
sucrose and lactose is a general property of bacterial levansucrases. may be produced from crude cellular extracts without costly
Levansucrase activity is frequently found in food fermenting lactic downstream processing (Vasiljevic & Jelen, 2001, 2003). Moreover,
acid bacteria, particularly L. reuteri, L. pontis and L. acidophilus. These GOS may be produced in situ during food fermentations or by
organisms have been successfully used for the generation of high using whey to produce food-grade GOS preparations. Bifidobacter-
levels of oligosaccharides in food fermentations (Tieking, Ehrmann, ium spp. deserve special attention as a relevant source of enzymes
Vogel, & Gänzle, 2005; Tieking et al., 2005). suitable for production of galacto-oligosaccharides. The competi-
tiveness of bifidobacteria in the intestinal tract of mammals is
5.6. Galacto-oligosaccharides linked to their ability to metabolize a large variety of oligo- and
polysaccharides. It was estimated that 8% of the genomic
Galacto-oligosaccharides (GOS) are commercially produced information in Bifidobacterium longum is dedicated to the
from lactose using fungal b-galactosidases. The b-galactosidases metabolism of complex carbohydrates (Ventura, van Sinderen,
 ARTICLE IN PRESS

692 M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694

Transgalactolysation Other acceptor molecules

Hydrolysis (Lactose as acceptor) (examples)

O
O O
Lactose O OH
O
O OH HO
O
O
O OH
O
Glucose
O H2N
OH
OH
O O
O O
O O HO
O O OH
E537 O OH

β (1-4) O O
H2O
O OH
O OH
O O OH
O O O Mannose, fructose
O
OH Galactose

Fig. 3. Schematic overview of the hydrolysis- and acceptor-reaction catalyzed by b-galactosidases. Carbohydrate moieties are drawn to indicate the carbon atoms and the
oxygen atoms of interest; galactose moieties are shaded gray. Lactose hydrolysis involved intermediate linkage of the galactosyl moiety to the glutamine residue in the
active site of the enzyme (E537 in the b-galactosidase of Escherichia coli) and subsequent transfer to H2O acting as galactosyl acceptor. Transgalactosylation to lactose or
galacto-oligosaccharides yields b(1-2), b(1-3), b(1-4) or b(1-6) linked galacto-oligosaccharides (the latter two predominating in most enzymes); other monosaccharides as
well as phenolic compounds have additionally been shown to act as galactosyl acceptor.

3.2 3.2
[Products] (%)

[Products] (%)

2.4 2.4

1.6 1.6

0.8 0.8

0.0 0.0
0 5 10 15 20 25 30 30 40 50 60
[Lactose ] (%) Temperature (°C)

Fig. 4. Lactose turnover by b-galactosidase in crude cellular extract from Lactobacillus delbrueckii spp. bulgaricus. Panel A: influence of lactose concentration on product
composition and Panel B: influence of temperature; K, glucose; ’, galactose; J, disaccharides; &, trisaccharides; D, tetrasaccharides. Replotted from Vasiljevic and Jelen
(2003).

Fitzgerald, & Zink, 2004). Several enzymes with activity on lactose biological as well as physico-chemical treatments are becoming
or galacto-oligosaccharides were characterized at a biochemical more complex, and (iv) available waste treatment scenarios are
level. The enzyme b-Gal-II from Bifidobacterium adolescentis becoming more expensive. Therefore, the desire to convert lactose
DSM20083 has a low transgalactosylation activity and a remark- and lactose-containing components into commercially viable
able specificity for hydrolysis of lactose, 40 galactosyllactose and products is increasing, and additional innovative applications in
higher b-(1-4) linked galacto-oligosaccharides (Hinz, van den the pharmaceutical and chemical industries are needed.
Broek, Beldman, Vincken, & Voragen, 2004). Bifidobacterium The traditional view of lactose as a commodity, produced by
infantis HL96 harbors two b-galactosidase genes, b-Gal-I and b- traditional crystallization or possibly other processes such as
GaI-III. Both enzymes are active towards lactose but the spray drying, has not been economically advantageous in the past.
oligosaccharide yield of b-Gal-I exceeded that of b-gal-III more Although there are several industrial lactose manufacturers
than threefold (Hung et al., 2001). offering a wide array of lactose products for diverse uses (from
infant foods to confectionery to pharma lactose), the overall
market for the traditional lactose products is relatively static and
6. Conclusions and future prospects radically new approaches to lactose utilization are needed to
achieve a ‘‘quantum leap’’ in converting this unique carbohydrate
Lactose is presently a most underutilized dairy component. The to new industrially attractive products. Some of the lactose
continued struggle to find new uses for the cheese whey is tied to derivatives discussed above are just a few examples of the new
finding new uses for lactose, the main whey component. It trends in surplus lactose management. Additional major oppor-
becomes increasingly difficult to dispose of whey or any other tunities may arise in using lactose (with or without prior
process streams containing high amounts of lactose from dairy hydrolysis) for various fermentation applications. Lactose used
and cheese operations as: (i) environmental discharge standards to be one of the cheapest fermentation carbohydrate on the
are getting higher and more difficult to comply with, (ii) market for some time. It is not inconceivable that lactose, in its
authorities are becoming more vigilant in all regions, (iii) crudest form as whey or UF permeate, could become an
 ARTICLE IN PRESS

M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694 693

economical source of biogas or bioethanol. Unfortunately, the Fox, P. F., & McSweeney, P. L. H. (1998). Dairy chemistry and biochemistry. London,
literature of the last 65 years is replete with similar suggestions UK: Blackie Academic and Professional.
Garman, J., Coolbear, J., & Smart, J. (1996). The effect of cations on the hydrolysis of
and pronouncements, while the dairy world is still waiting for the lactose and the transferase reactions catalysed by b-galactosidase from six
‘‘final solution’’ of the whey (and thus lactose) problem. strains of lactic acid bacteria. Applied Microbiology and Biotechnology, 46,
Finally, addressing the widespread problem of lactose intoler- 22–27.
Geciova, J., Giesova, M., & Jelen, P. (2002). Disruption of Streptococcus thermophilus
ance on a global level could bring major health benefits to many 143 culture by three mechanical methods for increased b-galactosidase
lactose intolerant populations around the world who presently activity. Milchwissenschaft, 57, 509–511.
have little access to milk and dairy products. This would also Geilman, W. G. (1993). Preparation and properties of syrups made by the hydrolysis of
lactose. Bulletin 289, pp. 33–37. Brussels, Belgium: International Dairy
benefit the industrial dairy processors in opening up new
Federation.
attractive markets for dried whey and other dairy products Gekas, V., & Lopez-Leiva, M. (1985). Hydrolysis of lactose—a literature review.
containing lactose. Such a development would bring about a Process Biochemistry, 20, 2–12.
recognition of milk as a truly ‘‘mother nature’s original nutraceu- Gerling, K. G. (1997). Large scale production of lactobionic acid—use and new
applications. In Whey (pp. 251–261). Brussels, Belgium: International Dairy
tical product’’, as well as converting lactose from money-losing Federation.
inconvenience of the past to money-making opportunity. Gibson, G. R., & Roberfroid, M. B. (1995). Dietary modulation of the human colonic
microbiota: Introducing the concept of prebiotics. Journal of Nutrition, 125,
1401–1412.
References Gillis, G. (1920). Nouvelles recherches sur le sucre de lait. Recueil des Travaux
Chimiques des Pays-Bas, 38, 88–125.
Haase, G., & Nickerson, T. A. (1966). Kinetic reactions of a- and b-lactose. I.
Albayrak, N., & Yang, S. T. (2002). Production of galacto-oligosaccharides from
Mutarotation. Journal of Dairy Science, 49, 127–132.
lactose by Aspergillus oryzae b-galactosidase immobilized on cotton cloth.
Haggett, T. O. R. (1976). The acid catalysed hydrolysis of lactose using a cation
Biotechnology and Bioengineering, 77, 8–19.
exchange resin. New Zealand Journal of Dairy Science and Technology, 11,
Als-Nielsen, B., Gluud, L. L., & Gluud, C. (2004). Non-absorbable disaccharides for
176–179.
hepatic encephalopathy: Systematic review of randomized trials. British
Haque, M. K., & Roos, Y. H. (2004). Water sorption and plasticization behavior
Medical Journal, 328, 1046–1050.
of spray-dried lactose/protein mixtures. Journal of Food Science, 69,
Anonymous. (1981). Corning and Kroger turn whey to yeast. Chemical and
E384–E391.
Engineering News, 59, 9.
Haque, M. K., & Roos, Y. H. (2005). Crystallization and X-ray diffraction of crystals
Anonymous. (1984). Immobilized enzymes and fermentation technologies com-
formed in water-plasticized amorphous spray-dried and freeze-dried lactose/
bine to produce baker’s yeast from whey. Food Technology, 38, 26–27.
protein mixtures. Journal of Food Science, 70, E359–E366.
Asp, N. G., Burvall, A., Dahlqvist, A., Hallgren, P., & Lundblad, A. (1980).
Harju, M. (1987a). Lactose hydrolysis. Bulletin 212, pp. 50–55. Brussels, Belgium:
Oligosaccharide formation during hydrolysis of lactose with Saccharomyces lactis
International Dairy Federation.
lactase (Maxilacts): Part 2-oligosaccharide structures. Food Chemistry, 5, 147–153.
Harju, M. (1987b). A method for the specific separation of lactose from skim milk.
Bhargava, A., & Jelen, P. (1996). Lactose solubility and crystal growth as affected by
Finnish Journal of Dairy Science, 45, 82–93.
mineral impurities. Journal of Food Science, 61, 180–184.
Herrington, B. L. (1934). Some physico-chemical properties of lactose IV. Journal of
Block, R. J. (1952). US Patent 2 592 509.
Dairy Science, 17, 659–670.
Böhm, G., Jelinek, J., Stahl, B., van Laere, K., Knol, J., Fanaro, S., et al. (2004).
Hestrin, S., & Avigad, G. (1958). The mechanisms of polysaccharide production
Prebiotics in infant formulas. Journal of Clinical Gastroenterology, 38, S76–S79.
from sucrose. Biochemical Journal, 69, 388–395.
Boon, M. A., Janssen, A. E. M., & van’t Riet, K. (2000). Effect of temperature and
Hinz, S. W. A., van den Broek, L. A. M., Beldman, G., Vincken, J.-P., & Voragen, A. G. J.
enzyme origin on the enzymatic synthesis of oligosaccharides. Enzyme and
(2004). b-Galactosidase from Bifidobacterium adolescentis DSM20083 prefers
Microbial Technolology, 26, 271–281. b(1,4) galactosides over lactose. Applied Microbiology and Biotechnology, 66,
Bouhnik, Y., Raskine, L., Simoneau, G., Vicaut, E., Neut, C., Flourié, B., et al. (2004). 276–284.
The capacity of nondigestible carbohydrates to stimulate fecal bifidobacteria in Holsinger, V. H. (1988). Lactose. In N. P. Wong, R. Jenness, M. Keeney, & E. H. Marth
healthy humans: A double-blind, randomized, placebo-controlled, parallel- (Eds.), Fundamentals of dairy chemistry, 3rd ed (pp. 279–342). New York, NY,
group, dose-response relation study. American Journal of Clinical Nutrition, 80, USA: Van Nostrand Reinhold Co.
1658–1664. Hopkins, M. J., & Macfarlane, G. T. (2003). Nondigestible oligosaccharides enhance
Bridiau, N., Taboubi, S., Marzouki, M., Legoy, M. D., & Maugard, T. (2006). b- bacterial colonization resistance against Clostridium difficile in vitro. Applied
Galactosidase catalyzed selective galactosylation of aromatic compounds. and Environmental Microbiology, 69, 1920–1927.
Biotechnology Progress, 22, 326–330. Huber, R. E., Kurz, G., & Wallenfels, K. (1976). A quantitation of the factors which
Bruins, M. E., Strubel, M., van Lieshout, J. F. T., Janssen, A. E. M., & Boom, R. M. affect the hydrolase and transgalactosylase activities of b-galactosidase (E. coli)
(2003). Oligosaccharide synthesis by the hyperthermostable b-glucosidase on lactose. Biochemistry, 15, 1994–2001.
from Purococcus furiosus: Kinetics and modeling. Enzyme and Microbial Hudson, C. S. (1908). Further studies on the forms of milk sugar. Journal of the
Technology, 33, 3–11. American Chemical Society, 30, 1767–1783.
Bury, D., & Jelen, P. (2000). Lactose hydrolysis using a disrupted dairy culture: Hung, M.-N., Xia, Z., Hu, N.-T., & Lee, B. H. (2001). Molecular and biochemical
Evaluation of technical and economical feasibility. Canadian Agricultural analysis of two b-galactosidases from Bifidobacterium infantis HL96. Applied
Engineering, 42, 75–80. and Environmental Microbiology, 67, 4256–4263.
Bury, D., Jelen, P., & Kalab, M. (2001). Disruption of Lactobacillus delbrueckii subsp. IDF. (1993). Lactose hydrolysis. Bulletin 289. Brussels, Belgium: International Dairy
bulgaricus 11842 cells for lactose hydrolysis in dairy products: A comparison of Federation, 71pp.
sonication, high-pressure homogenization and bead milling. Innovative Food IDF. (2005). World dairy situation. Bulletin 399. Brussels, Belgium: International
Science & Emerging Technologies, 2, 23–29. Dairy Federation, pp. 4 and 10.
Cheng, C.-C., Yu, M.-C., Cheng, T.-C., Sheu, D.-C., Duan, K.-J., & Tai, W.-L. Jelen, P., & Coulter, S. T. (1973a). Effects of supersaturation and temperature on the
(2006). Production of high-content galacto-oligosaccharide by enzyme growth of lactose crystals. Journal of Food Science, 38, 1182–1185.
catalysis and fermentation with Klyveromyces marxianus. Biotechnology Letters, Jelen, P., & Coulter, S. T. (1973b). Effects of certain salts and other whey substances
on the growth of lactose crystals. Journal of Food Science, 38, 1186–1189.
Cote, L., & Ahlgren, J. (1993). Metabolism in microorganisms. Part 1. Levan and Jelen, P., & Michel, C. (1999). Sensory impact in protein-standardised milk.
levansucrase. In Science and technology of fructans (pp. 141–161). Boca Raton FL, Milchwissenschaft, 54, 438–441.
USA: CRC Press. Jelen, P., & Tossavainen, O. (2004). Low lactose and lactose-free milk and dairy
Cummings, J. H., Macfarlane, G. T., & Englyst, H. N. (2001). Prebiotic digestion and products: Prospects, technologies and applications. Australian Journal of Dairy
fermentation. American Journal of Clinical Nutrition, 73, 415S–420S. Technology, 58, 161–165.
de Boer, R., & Robbertsen, T. A. (1981). A purified hydrolyzed lactose syrup made Jørgensen, F., Hansen, O. C., & Sougaard, P. (2001). High-efficiency synthesis of
from ultrafiltration permeate. Netherlands Milk and Dairy Journal, 35, 95–111. oligosaccharides with a truncated b-galactosidase from Bifidobacterium
Dills, W. L. (1989). Sugar alcohols as bulk sweeteners. Annual Reviews in Nutrition, bifidum. Applied Microbiology and Biotechnology, 57, 647–652.
9, 161–186. Jouppila, K., & Roos, Y. H. (1994). Glass transitions and crystallization in milk
Drapier-Beche, N., Fanni, J., & Parmentier, M. (1999). Physical and chemical powders. Journal of Dairy Science, 77, 2907–2915.
properties of molecular compounds of lactose. Journal of Dairy Science, 82, Jouppila, K., Kansikas, J., & Roos, Y. H. (1997). Glass transition, water plasticization,
2558–2563. and lactose crystallization in skim milk powder. Journal of Dairy Science, 80,
Elliott, A. J., Datta, N., Amenu, B., & Deeth, H. C. (2005). Heat-induced and other 3152–3160.
chemical changes in commercial UHT milks. Journal of Dairy Research, 72, 1–5. Kawade, T., Sakakibara, M., Nomura, A., Suzuki, K., & Kunugi, S. (1999). Effect of
EMEA. (2002). Risk and regulatory assessment of lactose and other products prepared pressure on free and immobilized b-galactosidase. Faculty of engineering
for using Calf Rennet. Biotechnology working party. The European Agency for research report (Vol. 39, pp. 45–54). Fukui University, Japan.
the Evaluation of Medical Products. Kouassi, K., Jouppila, K., & Roos, Y. H. (2002). Effects of k-carrageenan on
Englyst, K. N., & Englyst, H. N. (2005). Carbohydrate bioavailability. British Journal of crystallization and invertase activity in lactose-sucrose systems. Journal of
Nutrition, 84, 1–11. Food Science, 67, 2190–2195.
 ARTICLE IN PRESS

694 M.G. Gänzle et al. / International Dairy Journal 18 (2008) 685–694

Kozempel, M. F., Kurantz, M. J., Craig, J. C., & Hicks, K. B. (1995). Development of a Smart, J. B. (1988). Effect of whey components on the rate of crystallization and
continuous lactulose process: Separation and purification. Biotechnology solubility of a-lactose monohydrate. New Zealand Journal of Dairy Science and
Progress, 11, 592–595. Technology, 23, 275–289.
Kreft, M. E., & Jelen, P. (2000). Stability and activity of b-galactosidase in sonicated Smart, J. B. (1991). Transferase reactions of the b-galactosidase from Streptococcus
cultures of Lactobacillus delbrueckii subsp. bulgaricus 11842 as affected by thermophilus. Applied Microbiology and Biotechnology, 34, 495–501.
temperature and ionic environments. Journal of Food Science, 65, 1364–1368. Smart, J. B. (1993). Transferase reaction of b-galactosidases—new product opportu-
Kreft, M. E., Roth, L., & Jelen, P. (2001). Lactose hydrolysing ability of sonicated nities. Bulletin 289, pp. 16–22. Brussels, Belgium: International Dairy Federation.
cultures of Lactobacillus delbrueckii subsp. bulgaricus 11842. Le Lait, 81, Southard, J. H., & Belzer, F. O. (1995). Organ preservation. Annual Reviews in
355–364. Medicine, 46, 235–247.
Kunz, C., Rudloff, S., Baier, W., Klein, N., & Strobel, S. (2000). Oligosaccharides in Takada, M., Ogawa, K., Saito, S., Murata, T., & Usui, T. (1998). Chemo-enzymatic
human milk: Structural, functional and metabolic aspects. Annual Reviews in synthesis of galactosylmaltooligosaccharidonolactone as a substrate analogue
Nutrition, 20, 699–722. inhibitor for mammalian a-amylase. Journal of Biochemistry (Tokyo), 123,
Larhrib, H., Martin, G. P., Prime, D., & Marriott, C. (2003). Characterisation and 508–515.
deposition studies of engineered lactose crystals with potential for use as a Thurlby, J. S. (1976). Chrystallization Kinetics of Alpha Lactose. Journal of Food
carrier for aerosolised salbutamol sulfate from dry powder inhalers. European Science, 41, 38–42.
Journal of Pharmaceutical Sciences, 19, 211–221. Tieking, M., Ehrmann, M. A., Vogel, R. F., & Gänzle, M. G. (2005). Molecular and
Lee, Y. K. (1999). Handbook of probiotics. New York, NY, USA: Wiley. functional characterization of a levansucrase from Lactobacillus sanfranciscen-
Lee, Y. L., Kim, C. S., & Oh, D. K. (2004). Lactulose production by b-galactosidase in sis. Applied Microbiology and Biotechnology, 66, 655–663.
permeabilized cells of Kluyveromyces lactis. Applied Microbiology and Biotech- Tieking, M., & Gänzle, M. G. (2005). Exopolysaccharides from cereal-associated
nology, 64, 787–793. lactobacilli. Trends in Food Science and Technology, 16, 79–84.
MacBean, R., Hall, R., & Willman, N. (1979). Heterogeneous acid catalysed Tieking, M., Kühnl, W., & Gänzle, M. G. (2005). Evidence for formation of
hydrolysis of lactose with cation exchange resins. Australian Journal of Dairy heterooligosaccharides by Lactobacillus sanfranciscensis during growth in
Technology, 34, 53–59. wheat sourdough. Journal of Agricultural and Food Chemistry, 53, 2456–2461.
Matthews, B. W. (2005). The structure of E. coli b-galactosidase. Comptes Rendus Timmermans, E. (1997). Lactose derivatives: Functions and applications. In Whey
Biologies, 328, 549–556. (pp. 233–250). Brussels, Belgium: International Dairy Federation.
Mayer, J., Contrad, J., Klaiber, I., Lutz-Wahl, S., Beifuss, U., & Fischer, L. (2004). Topping, D. L., & Clifton, P. M. (2001). Short-chain fatty acids and human colonic
Enzymatic production and complete nuclear magnetic resonance assignment function: Roles of resistant starch and nonstarch polysaccharides. Physiological
of the sugar lactulose. Journal of Food and Agricultural Chemistry, 52, Reviews, 81, 1031–1064.
6983–6990. Twieg, W. C., & Nickerson, T. A. (1968). Kinetics of lactose crystallization. Journal of
Mehaia, M. A., Alvarez, J., & Cheryan, M. (1993). Hydrolysis of whey permeate Dairy Science, 51, 1720–1724.
lactose in a continuous stirred tank membrane reactor. International Dairy van Hijum, S. A. F. T., Kralj, S., Ozimek, L. K., Kijkhuizen, L., & van Geel Schutten, I. G.
Journal, 3, 179–192. H. (2006). Structure–function relationships of glucansucrase and fructansu-
Miao, S., & Roos, Y. H. (2005). Crystallization kinetics and X-ray diffraction of crase enzymes from lactic acid bacteria. Microbiology and Molecular Biology
crystals formed in amorphous lactose, trehalose, and lactose/trehalose Review, 70, 157–176.
mixtures. Journal of Food Science, 70, E350–E358. Van Kreveld, A. (1969). Growth rates of lactose crystals in solutions of stable
Michaels, A. S., & van Kreveld, A. (1966). Influence of additives on growth rates in anhydrous a-lactose. Netherlands Milk and Dairy Journal, 23, 258–275.
lactose crystals. Netherlands Milk and Dairy Journal, 20, 163–181. Van Kreveld, A., & Michaels, A. S. (1965). Measurement of crystal growth of
Miyasato, M., & Ajisaka, K. (2004). Regioselectivity in b-galactosidase-catalyzed a-lactose. Journal of Dairy Science, 48, 259–265.
transglycosylation for the enzymatic assembly of D-galactosyl-D-mannose. Van Loo, J., Cummings, J., Delzenne, N., Englyst, H., Franck, A., Hopkins, M., et al.
Bioscience, Biotechnology and Biochemistry, 68, 2086–2090. (1999). Functional food properties of non-digestible oligosaccharides: A
Nasirpour, A., Scher, J., Lindner, M., & Desobry, S. (2006). Modeling of lactose consensus report from the ENDO project (DGXII AIRII-CT94-1095). British
crystallization and colour changes in model infant foods. Journal of Dairy Journal of Nutrition, 81, 121–132.
Science, 89, 2365–2373. Vasiljevic, T., & Jelen, P. (2001). Production of b-galactosidase for lactose hydrolysis
Nickerson, T. A. (1962). Lactose crystallization in ice cream. IV. Factors responsible in milk and dairy products using thermophilic lactic acid bacteria. Innovative
for reduced incidence of sandiness. Journal of Dairy Science, 45, 354–359. Food Science and Emerging Technologies, 2, 75–85.
Nickerson, T. A. (1974). Lactose. In B. H. Webb, A. H. Johnson, & J. A. Alford (Eds.), Vasiljevic, T., & Jelen, P. (2002). Lactose hydrolysis in milk as affected by
Fundamentals of Dairy Chemistry (2nd ed.), (pp. 273–324). Westport, CA, USA: neutralizers used for preparation of crude b-galactosidase extracts from
AVI Publishing. Lactobacillus delbrueckii subsp. bulgaricus 11842. Innovative Food Science &
Nickerson, T. A. (1979). Lactose chemistry. Journal of Agricultural and Food Emerging Technologies, 3, 175–184.
Chemistry, 27, 672–677. Vasiljevic, T., & Jelen, P. (2003). Oligosaccharide production and proteolysis during
Nickerson, T. A., & Moore, E. E. (1974a). Alpha lactose and crystallization rate. lactose hydrolysis using crude cellular extracts from lactic acid bacteria. Le Lait,
Journal of Dairy Science, 57, 160–164. 83, 453–467.
Nickerson, T. A., & Moore, E. E. (1974b). Lactose influencing lactose crystallization. Vasiljevic, T., Wismer, W., & Jelen, P. (2003). Sensory effect of lactose hydrolysis in
Journal of Dairy Science, 57, 1315–1319. milk by crude cellular extracts from Lactobacillus delbrueckii ssp. bulgaricus
Nilsson, U., & Nyman, M. (2005). Short-chain fatty acid formation in the hindgut of 11842. Milchwissenschaft, 58, 167–170.
rats fed oligosaccharides varying in monomeric composition, degree of Veereman-Wauters, G. (2005). Application of prebiotics in infant foods. British
polymerisation and solubility. British Journal of Nutrition, 94, 705–713. Journal of Nutrition, 93, S57–S60.
Ogata, Y., Fujita, K., Ishigami, H., Hara, K., Terada, A., Hara, H., Fujimori, I., & Ventura, M., van Sinderen, D., Fitzgerald, G. F., & Zink, R. (2004). Insights into the
Misuoka, T. (1993). Effect of a small amount ot 4G-beta-D-galactosylsucrose taxonomy, genetics and physiology of bifidobacteria. Antonie van Leeuwenhoek,
(lactosucrose) on fecal flora and fecal properties. Journal of Japanese Society of 86, 205–223.
Nutrition and Food Science, 46, 317–323. Visser, R. A. (1983). Crystal growth kinetics of alpha-lactose hydrate. Ph.D. Thesis,
Ohtsuka, K., Hino, S., Fukushima, T., Ozawa, O., Kanematsu, T., & Uchida, T. (1992). University of Nijmegen, The Netherlands, 142pp.
Characterization of levansucrase from Rahnella aquatilis JCM-1683. Bioscience Wallenfels, K., & Weil, R. (1972). b-Galactosidase. In P. D. Boyer (Ed.), The enzymes,
Biotechnology Biochemistry, 56, 1373–1377. Vol. 7 (pp. 617–663). New York, NY, USA: Acad. Press.
Onishi, N., Yamashiro, A., & Yokozeki, K. (1995). Production of galacto-oligosac- Whittier, E. O. (1925). Lactose. Chemical Reviews, 2, 85–125.
chyaride from lactose by Sterigmatomyces elviae CBS8119. Applied and Whittier, E. O. (1944). Lactose and its utilization—a review. Journal of Dairy Science,
Environmental Microbiology, 61, 4022–4025. 27, 505–537.
Patel, K. N., & Nickerson, T. A. (1970). Influence of sucrose on the mutarotation Yanahira, S., Kobayashi, T., Suguri, T., Nakakoshi, M., Miura, S., Ishikawa, H.,
velocity of lactose. Journal of Dairy Science, 53, 1654–1958. et al. (1995). Formation of oligosaccharides from lactose by Bacillus
Paterson, A. H. J., Brooks, G. F., Bronlund, J. E., & Foster, K. D. (2005). Development of circulans b-galactosidase. Bioscience, Biotechnology and Biochemistry, 59,
stickiness in amorphous lactose at constant T–Tg levels. International Dairy 1021–1026.
Journal, 15, 513–519. Yanahira, S., Yabe, Y., Nakakoshi, M., Miura, S., Matsubara, N., & Ishikawa, H. (1998).
Pritzwald-Stegman, B. F. (1986). Lactose and some of its derivatives. Journal of the Structures of novel acidic galactooligosaccharides synthesized by Bacillus
Society of Dairy Technology, 39, 91–97. circulans b-galactosidase. Bioscience, Biotechnology and Biochemistry, 62,
Roos, Y., & Karel, M. (1992). Crystallization of Amorphous Lactose. Journal of Food 1791–1794.
Science, 57, 775–777. Zadow, J. G. (1984). Lactose: Properties and uses. Journal of Dairy Science, 67,
Ryder, D. N. (1988). Hydrolysis of lactose in whey products. Bulletin 202, pp. 45–52. 2654–2679.
Brussels, Belgium: International Dairy Federation. Zadow, J. G. (1993). Economic considerations related to the production of lactose and
Satory, M., Führlinger, M., Haltrich, D., Kulbe, K. D., Pittner, F., & Nidetzky, B. (1997). lactose by-products. Bulletin 289, pp. 10–15. Brussels, Belgium: International
Continuous enzymatic production of lactobionic acid using glucose-fructose Dairy Federation.
oxidoreductase in an ultrafiltration membrane reactior. Biotechnology Letters, Zeng, X. M., Martin, G. P., Marriott, C., & Pritchard, J. (2000a). Crystallization of
19, 1205–1208. lactose from carbopol gels. Pharmaceutical Research, 17, 879–886.
Shukla, T. P. (1975). b-Galactosidase technology—a solution to the lactose problem. Zeng, X. M., Martin, G. P., Marriott, C., & Pritchard, J. (2000b). The influence of
CRC Critical Reviews in Food Technology, 5, 325–356. crystallization conditions on the morphology of lactose intended for use as a
Sienkiewicz, T., & Riedel, C.-L. (1990). Whey and whey utilization (2nd ed.). carrier for dry powder aerosols. Journal of Pharmacy and Pharmacology, 52,
Gelsenkirchen-Buer, Germany: Verlag Th. Mann (pp. 145; 263–264). 633–652.