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Carbohydrate Preferences of Bifidobacterium Species-Isolated

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Carbohydrate Preferences of Bifidobacterium Species-Isolated Curr. Issues Intest. Microbiol. (2003) 4: 71-75. Carbohydrate Preferences of Bifidobacterium Species 71 Carbohydrate Preferences of Bifidobacterium Species Isolated from the Human Gut Richard J. Palframan, Glenn R. Gibson (De Vriers, 1967) and are therefore unable to carry out the and Robert A. Rastall* usual glycolysis pathway or the hexose monophosphate shunt pathway. The bifidus pathway depends on the Food Microbial Sciences Unit, School of Food Biosciences, presence of fructose-6-phosphate phosphoketolase The University of Reading, PO Box 226, Whiteknights, (F6PPK) (Fandi et al., 2001). The formation of acetate and Reading, RG6 6AP lactate as end products of this pathway are significant in terms of the health benefits of probiotics. In some species of bifidobacteria however, pyruvate is converted into formic Abstract acid and ethanol, yielding an extra ATP from 1 mole of glucose The presence of the Bifidus pathway allows The growth of nine species of Bifidobacterium on bifidobacteria to produce more ATP from carbohydrates media containing glucose, xylose, xylo- than conventional hetero and homofermentative pathways. oligosaccharides (XOS), xylan or fructo- The bifidus pathway yields 2.5 ATP from 1 mole of glucose, oligosaccharides (FOS) as the sole carbon source were as well as theoretically 1.5 moles of acetate and 1 mole compared in pure culture. The bifidobacteria differed lactate. These short chain fatty acids play a part in the in fermentation profiles when tested on different purported health promoting properties associated with carbohydrates. All species grew to their highest final prebiotics. It is known that acetate produced in the gut is optical density (OD) on a glucose containing medium, transported to the liver where it is utilised in the formation with the exception of B. catenulatum which of ATP. Lactate is a known to possess anti-microbial activity demonstrated a preference for xylose over glucose, which is active against a number of potentially pathogenic and XOS over FOS. B. bifidum grew to the highest OD bacteria (Roberfroid et al., 1995). on XOS compared to xylose suggesting a specific In order to develop new and improved prebiotic transport system for the oligosaccharide over the carbohydrates, it is imperative that we fully understand the monomer. This is consistent with a lack of β-xylosidase fermentation pathways and uptake mechanisms that activity present in the culture medium. Lactate, formate operate in the target organisms i.e. bifidobacteria and and acetate levels were determined and the ratios of lactobacilli. these metabolites altered between and within species In this study, the fermentation of a number of growing on different carbohydrates. In general, high carbohydrates by nine bifidobacteria species was lactate production correlated with low formate examined. The carbohydrates tested were xylose, XOS production and low lactate concentrations were and xylan with FOS and glucose as controls. These obtained at higher levels of formate. Bifidobacteria may carbohydrates were chosen because the fermentation of alter their metabolic pathways based upon the pentose sugars i.e. xylose, are poorly understood in carbohydrates that are available for their use. Bifidobacterium Spp. As xylooligosaccharides are emerging as prebiotics the understanding of their fermentation by Introduction bifidobacteria is of importance. XOS have a structure of [β-Xyl- (1-4)-]n, with a chain length between 2 and 9, with xylan having the same structure but a higher molecular Prebiotics such as fructooligosaccharides (Gibson, 1999) weight. and xylooligosaccharides (Vazquez et al., 2000) have been The highest optical density (OD) was used to shown to elicit a prebiotic effect both in vitro and in vivo. determine the carbohydrate preferences of each species Prebiotics are defined as “non-digestible food ingredients tested. In order to examine the biochemical activities of that stimulate the growth and/or the activity of one or a the species, high performance liquid chromatography limited number of bacteria in the colon, thereby improving (HPLC) analysis of acetate, lactate and formate the host health” (Gibson and Roberfroid, 1995). concentrations were also carried out. The xylosidase Prebiotics selectively stimulate the growth of activity of the species able to ferment XOS was also commensal probiotic bacteria in the colon due to their analysed using a β-xylosidase assay, this generated fermentation profiles. Species from the genus information about the location of xylose and XOS transport Bifidobacterium are the main target for these prebiotics mechanisms within each species. and numerous clinical trials have demonstrated the bifidogenic nature of prebiotics (Gibson et al., 1995). Materials and Methods Bifidobacteria differ from other colonic genera in their fermentation of carbohydrates. They lack the enzymes aldolase and glucose-6-phosphate NADP+ oxidoreductase Bacterial Growth Media Bacteria were grown on basal medium containing (g/l): peptone water (Oxoid Ltd. Basingstoke, UK) 2, yeast extract *For correspondence. Email [email protected]. © 2003 Horizon Scientific Press Further Reading Caister Academic Press is a leading academic publisher of advanced texts in microbiology, molecular biology and medical research. 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BlankBlank a) BlankBlank b) GlucoseGlu c ose *1 GlucoseGlu co se *1 XylanXy lan XylanXylan XyloseXy lose XyloseXy lose *2 XOSXOS *2 XOSXOS FOSFOS *1 FOSFOS *2 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 BlankBlank c) BlankBlank d) GlucoseGlu c ose *1 GlucoseGlu co se *1 XylanXy lan XylanXylan XyloseXy lose XyloseXy lose XOSXOS XOSXOS FOSFOS *1 FOSFOS *1 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 BlankBlank e) BlankBlank f) GlucoseGlu c ose *1 GlucoseGlu co se *1 XylanXy lan XylanXylan XyloseXy lose *2 XyloseXy lose *2 XOSXOS *2 XOSXOS FOSFOS *1 FOSFOS *2 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 BlankBlank g) BlankBlank h) GlucoseGlu c ose *2 GlucoseGlu co se *1 XylanXy lan *1 XylanXylan XyloseXy lose XyloseXy lose XOSXOS XOSXOS FOSFOS FOSFOS *1 0.00 0.20 0.40 0.60 0.80 1.00 1.20 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 FiFiguregure 1.1, Maximum Maximum OD values OD obtained va lues for obtained each test species for BlankBlank i) (n=3). eaca) hBifidobacterium test bifidums pecies (n=3). a) GlucoseGlu c ose *1 Bifidobacterium.bifidum,b) B. pseudolongum XylanXy lan *2 c) B. angulatum b)d) B. B. infantispseudolongum, c) B.angulatum, d) XyloseXy lose *1 B.infane) B. adolesentistis, f) B.
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