Review : Role of Galactooligosaccharides as Prebiotic

Abstract

Specific types of dietary carbohydrates, viz. Non-digestible oligosaccharides (NDOs), are known to pass through upper GI tract in intact or non-digestible form and promote the growth of beneficial bacteria in the colon. The acceleration of the industrial production of NDOs since the 1970s has mainly been achieved. Ability of galactooligosaccharide (GOS) to resists digestion and absorption in the small intestine and reaches the cecum and colon, where it is fermented by colonic bacteria. Because of bifidogenic nature and immuno-modulatory properties, GOS along with probiotics can ferment short chain fatty acids (SCFA) and gases, which is effective in genotoxicity, lowering cholesterol, improving infant health and efficient in diseases.

Full text

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Review : Role of Galactooligosaccharides as Prebiotic
M. N. Hingu,1 H.S.Shah,2
Dairy Science College, Amreli1, Gujarat, India
Department of Microbiology, Shri A. N. Patel P. G. Institute, Anand2, Gujarat, India
Email (Corresponding Author1):- hingumitesh@gmail.com
Abstract:
Specific types of dietary carbohydrates, viz. Non-digestible oligosaccharides (NDOs), are
known to pass through upper GI tract in intact or non-digestible form and promote the growth of
beneficial bacteria in the colon. The acceleration of the industrial production of NDOs since the
1970s has mainly been achieved. Ability of galactooligosaccharide (GOS) to resists digestion and
absorption in the small intestine and reaches the cecum and colon, where it is fermented by
colonic bacteria. Because of bifidogenic nature and immuno-modulatory properties, GOS along
with probiotics can ferment short chain fatty acids (SCFA) and gases, which is effective in
genotoxicity, lowering cholesterol, improving infant health and efficient in diseases.
Keywords: Galactooligosaccharide (GOS), Prebiotic, Probiotic, Non digestible oligosaccharides
1. Introduction:
The term “prebiotic” was coined by Prof. Glenn Gibson and Prof. Marcel Roberfroid in
1995 and defined it as a “non-digestible food ingredient that beneficially affects the host by
selectively stimulating the growth and/or activity of one or a limited number of bacteria in the
colon, and thus improves host health” (Gibson and Roberfroid, 1995). Specific types of dietary
carbohydrates, viz. non-digestible oligosaccharides (NDOs), are known to pass through upper GI
tract in intact or non-digestible form and promote the growth of beneficial bacteria in the colon,
and are thus recognized as Prebiotics (Fuller and Gibson, 1997). Milk is a classical natural
example of a „prebiotic diet‟ of mammals during infancy. The galactooligosaccharides (GOS)
present in milk, are the most relevant component for the prebiotic effect of human milk (Kunz et
al., 2000; Boehm and Stahl, 2007). As an alternative to probiotics, prebiotics can also modulate
the gut microbiota (Bhatia and Rani, 2007). Both popular concepts target the gastrointestinal
microbiota (Venter, 2007).
2. Production and charecterization of NDOs:

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Recent data on GOS production are not available. However, the market for prebiotics in food
is growing rapidly. A 2007 report on the world prebiotic market states that there are over 400
prebiotic food products and more than 20 companies producing oligosaccharides and fibres used
as prebiotics [http://www.ubic-consulting.com/template/fs/The-World-Prebiotic-Ingredient-
Market.pdf]. A Frost & Sullivan review reported that the European prebiotics market is currently
worth €87 million, and will reach €179.7 million by 2010. Global sales were approximately $1
billion USD in 2011; and GIA (Global Industry Analyst) predicts that figure will quintuple by
2018 (http://www.foodproductiondaily.com/Financial/Prebiotics-market-to-hit-4.8-billion-by-
2018).
The acceleration of the industrial production of NDOs since the 1970s has mainly been
achieved by the development of downstream processes using enzymatic and chemical reactions.
Today, over 20 different types of NDOs are on the world market, which are either extracted from
natural sources (e.g. raffinose and soybean oligosaccharides), obtained by the enzymatic
hydrolysis of polysaccharides (e.g. xylooligosaccharides and isomaltooligosaccharides), or
produced by enzymatic transglycosylation (e.g. GOS, and fructooligosaccharides-FOS). The most
abundantly supplied and utilized group of NDOs as food ingredients are GOS and FOS, which are
generally produced by enzymatic transglycosylation because of adequate supply of the raw
materials and the high efficiency of the reaction (Sako et al., 1999).
3. Galactooligosaccharides (GOS)
3.1. Technolgical Characteristics of Galactooligosaccharides
3.1.1 Production:
Galactose containing oligosaccharides of the form Glu α 1-4[β Gal 1-6]n where n=2-5, are
termed galactooligosaccharides and are present in both human as well as cow milk (Tuohy et al.,
2005). GOS is produced from lactose by the action of β-galactosidases having
transgalactosylation activity (Akiyama et al., 2001; Aslan and Tanrıseven, 2007) and the GOS
production can be enhanced by mixing glucose oxidase and β-galactosidase (Cheng-Chao et al.,
2006). One or more d-galactosyl units onto the D-galactose moiety of lactose used to produce
GOS by β–galactosidases during the hydrolysis of β–galactoside linkage of lactose (Moller et al.,
2001). The linkage between the galactose units, the efficiency of transgalactosylation, and the
components in the final products depend on the enzymes and the conditions used in the reaction
(Van Laere et al., 2000). The β -galactosidases used in different experiments are derived from

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various microbes viz. Bacillus circulans (Mozaffar et al., 1984), Bifidobacterium bifidum (Goulas
et al., 2007), Escherichia coli (Chen et al., 2003), Bullera singularis (Shin et al., 1998) and
Archaeon Sulfolobus solfataricus (Splechtna et al., 2001). Thermostable or immobilized β-
galactosidase is generally used to increase the production of GOS (Smart, 1991 and Cheng et al.,
2006). Goulas and co-workers (2007) reported that the toluene treated whole cells of
Bifidobacterium bifidum NCIMB 41171 showed better GOS conversion from lactose (80-85%).
3.1.2 Physiological characteristics:
Commercially available GOS are mixtures of several molecular species of
oligosaccharides (more than 55%), lactose (~20%), glucose (~20%), and a small amount of
galactose. GOS are available in liquid and powder forms (Voragen, 1998).
4. Effect of Galactooligosaccharides (GOS) on human health
4.1 Indigestibility and Energy Value
GOS having β -configuration, whereas human gastrointestinal digestive enzymes are
mostly specific for α-glycosidic bonds and the activity of β-galactosidase localized at the brush
border membrane of the small intestine, which has the potential to digest GOS, is usually weak or
often deficient (Ito and Kimura, 1993), so it resists digestion and absorption in the small intestine
and reaches the cecum and colon, where it is fermented by colonic bacteria (Ishikawa et al.,
1995). The indigestibility of GOS in vivo has been demonstrated by means of the hydrogen breath
test (Tanaka et al., 1983; Ishikawa et al., 1995). According to the standardized method developed
in Japan (Oku, 1996), the caloric value of GOS is 1.73 kcal g-1 (Watanuki et al., 1996).
4.2 Effect of GOS on intestinal microbial ecology and host
4. 2. 1 Utilization of 4’-galactosyllactose by intestinal bacteria in vitro:
Bacteria which inhabit the large intestine may have acquired divergent glycolytic activities
to efficiently utilize non-digestible carbohydrates which are abundant in the colon (Iino and
Morishita, 1990). GOS can be fermented by some strains of intestinal Bifidobacteria,
Lactobacillus, Bacteroides, and Clostridium (Tanaka et al., 1983; Ishikawa et al., 1995;
Matsumoto, 1990).
4. 2. 2 Effects of indigestion on intestinal microflora
(i) Bifidogenic activity:

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GOS is bifidogenic in nature (Olano-Martin et al., 2002) and also effective at increasing
numbers of Bifidobacteria (Ito et al., 1990; Bouhnik et al., 1997, 1999 and 2007; Djouzi and
Andlueux, 1997; Rivero and Santamaría, 2001; Tzortzis et al., 2005; Macfarlane et al., 2006 and
Depeint et. al., 2013) and lactate whilst generating the least gas (Rycroft et al., 2001), which
exchanges the predominant bacteria in the healthy adult human intestine along with reduction in
breath hydrogen (Bouhnik et al., 1997).
(ii) Metabolism in the colon
Oligosaccharides may directly inhibit infections by enteric pathogens due to their ability to
act as structural mimics of the pathogen binding sites that coat the surface of GI epithelial cells
(Shoaf et al., 2006), also change the composition of the mucosa associated flora significantly
(Langlands et al., 2004), hence the effectiveness of a prebiotic depends on its ability to be
selectively fermented by and to support growth of specific targeted organisms (Huebner et al.,
2007). The end products of fermentation of carbohydrates by colonic bacteria are short chain fatty
acids (SCFA) and gases (Bouhnik et al., 1997; Pereira et al., 2003). The in vitro fermentation of
GOS showed highest decrease in numbers of Clostridia and higher amount of SCFAs (Rycroft et
al., 2001). In another study, GOS and other prebiotics reduced the adherence Enteropathogenic
Escherichia coli on tissue culture cells (Shoaf et al., 2006). It is having laxative effect too (Alles
et al., 1996).
4. 2. 3 Physiological effects of Galactooligosaccharides (GOS)
(i) Improvement of Defecation and Elimination of Ammonia:
GOS showed improved defecation (Deguchi et al., 1997) and the SCFA production
contributed by increasing osmotic pressure and stimulating peristalsis (Ishikawa et al., 1995).
Bifidobacteria have the ability to assimilate ammonia as a nitrogen source, hence it reduces the
blood ammonia and suppress the ammonia-producing bacteria. The assimilation of amino acids by
intestinal bacteria yields also SCFA and/or branched- chain fatty acids as well as ammonia as
fermentation products (Deguchi et al., 1993; Mwenya et al., 2005).
(II) Stimulation of Mineral Absorption and Bone Mineralization:
GOS stimulates the calcium and magnesium absorption, which takes place in both the
small and large intestines, and accompanied by a reduction in cecal pH and increase in cecal and
cecal digesta weight (Chonan and Watanuki, 1995). GOS when fed to rats (Chonan et al., 2001),
postmenopausal women (van den Heuvel et al., 2000) and young male volunteers (van den
Heuval et al., 1998) showed increased calcium, magnesium and iron absorption. It was shown that

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prebiotics stimulated the absorption of iron and of bone-relevant minerals such as calcium,
magnesium, and zinc in short-term experiments and improved bone mineral content in the long-
term perspective (Abrahms et al. 2005; Scholz-Ahrens et al., 2007). Trinidad and co-workers
(1996) demonstrated that acetic acid and propionic acid when introduced into the six human
volunteers distal colon directly; increased calcium disappearance in the colon.
(iii) Effect on Bacterial enzyme activities associated with genotoxicity and cancer:
Several bacterial enzymes, such as β-glucuronidase, β-glucosidase, and nitroreductase may
play a role in colon carcinogenesis by converting pre-carcinogens to proximal carcinogens
(Rowland, 1988). In human studies, Ishikawa et al., (1995) detected a decreasing tendency in
fecal β-glucuronidase activity at a dose of 10 g GOS per day. GOS (10%, w/w) was associated
with decreases in β-glucuronidase activity and secondary bile acid and increased β-galactosidase
concentrations in feces of human flora-associated rats (Kikuchi et al., 1996). The daily intake of
15 g of GOS in young healthy volunteers resulted in a significant increase in fecal acetate and a
significant decrease in fecal β-glucuronidase activity (van Dokkum et al., 1999). In vitro studies
using a three-stage continuous culture of the large gut (McBain and Macfarlane, 2001) indicated
that 5% GOS (85% pure), strongly suppressed bacterial synthesis of arylsulphatase, β-glucosidase
and β-glucuronidase in all three culture vessels. Butyrate also play an important role in preventing
cancer (Topping and Clifton, 2001).
(iv) Effect on cholesterol and lipid metabolism:
The hypotriglyceridemic and Hypocholesterolemic effects of NDOs may be attributable to
the reduction of hepatic synthesis and/or absorption of triglycerides and cholesterol (Kok et al.,
1996). The mechanisms by which dietary carbohydrates elicit their lipid-lowering effects are,
however, still a matter of debate. Limited numbers of reports are available which indicate positive
effects of GOS on serum cholesterol metabolism in human subjects. Chonan et al., (1995)
reported increase in LDL cholesterol and triglycerides with a striking reduction in HDL
cholesterol after consumption of GOS in postmenopausal status of the rats.
(v) Neonates and infants:
Babies provide a unique opportunity to investigate the effects of prebiotics on the gut
microbiota, as they are born with what is an essentially sterile GI tract. For the first 2 years of life,
bifidobacteria predominate in the colonic microbiota in breast-fed infants, constituting up to 95%
of culturable bacteria (Tissier, 1900 and 1906; Bullen and Willis, 1971). The majority of trials
have focused on demonstrating the abilities of oligosaccharides to increase faecal bifidobacteria
populations (Moro and Arslanoglu, 2005), and relatively few studies have been made on disease

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prevention and on hypersensitivity response. GOS alone or along with probiotics showed
reduction in eczema, atopic eczema, diarrhoea and dysentery (Sazwal et al., 2004) as well as IgE
associated disease (Kukkonen et al., 2007). Despite a few studies at present, the failure to find a
satisfactory mechanism of action implies that more investigations will be needed before prebiotics
can begin to play a role in the routine management of atopic disease (Macfarlane et al,. 2008).
Bakker-Zierikzee et al., (2005), fed the prebiotic mixture of GOS/FOS (0.6%) to 19 infants,
showed higher faecal acetate contentand also resulted in a similar effect on metabolic activity of
the flora as in breast-fed infants.
(vi) Effects on diseases:
Obesity and diet have long been linked to atherosclerosis, cardiovascular disease, type 2
diabetes (Wong et al., 1989; Singh et al., 1992), rheumatoid arthritis (Feldmann et al., 1996) and
in some inflammatory diseases (Gross et al., 1992; Woywodt et al. 1999). GOS shows dose-
dependent benefits in erythema and swelling of limbs, as well as histological findings in the hind
paw joints (Abe et al., 2004). Song and his co-workers (2013), found that GOS and GOS-rich
prebiotic yogurt when administered to SOD1G93A mice, showed significant delay in the disease
Amyotrophic lateral sclerosis (ALS) onset and prolonged the lifespan in SOD1G93A mice and also,
these products increased the concentration of folate, VitB12. Trans-galactooligosaccharide
significantly enhanced faecal bifidobacteria (3.5 g/d P < 0.005; 7 g/d P < 0.001) (Silk D. B. et. al.
2009).
4. 2. 4 Immunomodulatory Properties
(i) Prebiotics and immune system:
Immunity comprises both innate and adaptive responses (Albers et al., 2005). The gut
contains a major part of the body‟s immune system, termed the gut-associated lymphoid tissue
(GALT). To date, few studies have been made on interactions between fermentable carbohydrates
and the immune system, or whether they exert direct or indirect modulatory effects. Schley and
Field (2002) reviewed immunological effects that have been observed after adding prebiotics to
the diet of dogs/mice/rats. The effects includes increase in mucosal immunoglobulin production,
mesenteric lymph nodes, Peyer‟s patches, and altered cytokine formation and lymphocyte
numbers in the spleen and intestinal mucosa. In the conclusion they suggested that the methods by
which prebiotics can exert their effects on the immune system, and attenuate inflammation in the
colon, include increased SCFA production and increases in immunogenic bacteria such as
lactobacilli and bifidobacteria. The SCFAs stimulates apoptosis (Rowland, 1988) and may be a
protective factor in carcinogenesis (Scheppach and Weiler 2004), propionate has been shown to

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be anti-inflammatory with respect to colon cancer cells (Nurmi et al., 2005). Glutamine is used by
immune cells in the body and increased production of butyrate may reduce the requirement of
epithelial cells in the gut for this amino acid, thereby enhancing immune system reactivity
(Jenkins et al., 1999). Administration of galactooligosaccharide mixture [Bi2muno (B-GOS)] to
overweight adults (n=45) showed positive effects on the composition of the gut microbiota, the
immune response, and insulin, total cholesterol, and trans- galactooligosaccharide concentrations
(Vulveic et. al., 2013). They have also mentioned that B-GOS may be a useful candidate for the
enhancement of gastrointestinal health, immune function, and the reduction of metabolic
syndrome risk factors in overweight adults.
(ii) The mucosal immune system:
Bakker-Zierikzee et al., (2006), fed the prebiotic mixture of GOS/FOS (0.6%) to 19
infants, which showed higher faecal SIgA level in the body. In another study (Scholtens et al.,
2008) 215 infants were fed GOS/FOS containing formula and they also found the higher level of
SIgA in the body. The intestinal mucosa contains large amounts of sIgA which has a protective
role against adherence and invasion by harmful bacteria and viruses (Yasui et al., 1991).
(iii) Allergy:
Allergic diseases are associated foods and environmental substances (Ozaki et al., 2007).
Atopic dermatitis (AD) is one of the early signs of allergy in infancy, and can affect 10–25% of
children in Western countries and prebiotics shows the beneficial effects on the same (Moro et al.,
2006). In one study, infants (n =461) with high risk of allergy, when fed with GOS alond with
probiotics showed significant reduction in eczema and atopic eczema (Kukkonen et al., 2007).
Administration of GOS reduced the allergic airway eosinophilia in ovalbumin-sensitizedbrown
Norway rats (Sonoyama et al., 2005).
(iv) Inflammatory bowel diseases (IBD):
Ulcerative colitis (UC) and Crohn‟s disease (CD) are both inflammatory conditions that
are thought to result from inappropriate immune responses to the normal commensal gut
microbiota (Cummings et al., 2003). Mucosa-associated lymphoid tissue normally maintains gut
integrity by tightly regulating immune responses to commensal and pathogenic bacteria and
dietary antigens, by a balance of pro-inflammatory and anti-inflammatory cytokines. Studies
involving the use of prebiotics in human IBD are distinctly lacking (Cummings et al., 2003;
Macfarlane et al., 2004). In one report by Holma et al., (2002), rats were fed 4 g kg -1 body mass
of either whey or lactose-derived GOS per day shows increase in bifidobacterial numbers in the
animals, there was no reduction in inflammatory processes.

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5. Applications of Galactooligosaccharides (GOS) as food ingredients
In the 1980s, consumers' demand for healthy foods led to a dramatic increase in the
development of NDO-containing foods. Low carcinogenicity, low calorific values, and low
sweetness were the most important characteristics of these oligosaccharides which show
beneficial effects on human health. At the same time, NDOs generally have preferable
physicochemical characteristics applicable to various processed foods, fermented milk products,
breads, jams, confectionery, beverages, also used in baby foods and specialized foods for elderly
and hospitalized people. In particular, the stability of GOS in acidic and high-temperature
conditions enable GOS to be applied without decomposition in a wider variety of foods
(Macfarlane et al., 2008).
6. Safety aspects
GOS have a generally recognized as safe (GRAS) status due to the fact that they are
components of human milk and traditional yoghurt and they are produced from ingested lactose
by resident intestinal bacteria which produce β -galactosidase. It is non toxic and non mutagenic.
The only adverse effect of GOS known so far is transient diarrhoea due to the so-called „osmotic
diarrhoea', which occurs when excess GOS are consumed (Macfarlane et al., 2008).
7. New vision in the future prospects of GOS : Conclusion
Recent advances in the research of the intestinal microflora clarify the importance of host
microbial interaction for human health. The prebiotic concept borne from the preferential growth
stimulating activity of food components on specific types of colonic bacteria is being accepted
from a scientific point of view as well as from a consumer point of view. NDOs including GOS
are recognized as bifidogenic factors. However, much still should have to be clarified about the
intestinal microflora and host microbe interaction. Modern biotechnology techniques such as
detection and identification methods, molecular dissection of bacteria, and in vitro cell culture
methods will promote a better understanding of the role of NDOs and intestinal bacteria. GOS
seem to have several unique characteristics which enable manufactures to utilize GOS in various
foods. Scientific substantiation of the biological effects is necessary to make this a reality
(Macfarlane et al., 2008).
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