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Synthesis of prebiotic alpha- and beta-galactooligosaccharides (GOS) using the whole cells of Bifidobacterium bifidum NCIMB 41171 was investigated. Determination of alpha- and beta-galactosidase activities showed them to be at 3 and 205 g(-1) of freeze dried biomass, respectively, and they increased to 5 and 344 U g(-1), respectively, when cells were treated with toluene. Starting with 450-500 mg mL(-1) lactose, maximum GOS concentrations were observed at 80-85% lactose conversions and the mixtures contained oligosaccharides (with a degree of polymerisation >= 3) at 77-109 mg mL(-1) and trans-galactosylated disaccharides between 85-115 mg mL(-1). The GOS yield values varied between 36% and 43%. An alpha-linked disaccharide was detected and its presence was confirmed by gas chromatography mass spectroscopy. Cells were re-used up to 8 times without changes in reaction times or the substrate conversions to GOS. Oligosaccharide synthesis was not inhibited by the presence of glucose or galactose. The mixtures were successfully purified from glucose (92% of glucose removed) by fermentation with Saccharomyces cerevisiae with no losses in the oligosaccharide content and only a small decrease on the galactose. (c) 2006 Elsevier Ltd. All rights reserved.

Prebiotics are nondigestible food ingredients that encourage proliferation of selected groups of the colonic microflora, thereby altering the composition toward a more beneficial community. In the present study, the prebiotic potential of a novel galactooligosaccharide (GOS) mixture, produced by the activity of galactosyltransferases from Bifidobacterium bifidum 41171 on lactose, was assessed in vitro and in a parallel continuous randomized pig trial. In situ fluorescent hybridization with 16S rRNA-targeted probes was used to investigate changes in total bacteria, bifidobacteria, lactobacilli, bacteroides, and Clostridium histolyticum group in response to supplementing the novel GOS mixture. In a 3-stage continuous culture system, the bifidobacterial numbers for the first 2 vessels, which represented the proximal and traverse colon, increased (P < 0.05) after the addition of the oligosaccharide mixture. In addition, the oligosaccharide mixture strongly inhibited the attachment of enterohepatic Escherichia coli (P < 0.01) and Salmonella enterica serotype Typhimurium (P < 0.01) to HT29 cells. Addition of the novel mixture at 4% (wt:wt) to a commercial diet increased the density of bifidobacteria (P < 0.001) and the acetate concentration (P < 0.001), and decreased the pH (P < 0.001) compared with the control diet and the control diet supplemented with inulin, suggesting a great prebiotic potential for the novel oligosaccharide mixture.

Batch syntheses of isomaltooligosaccharides (IMO) from sucrose, using the enzymes dextransucrase and dextranase were performed with the aim of understanding the reaction mechanism and the parameters which affect product characteristics and molecular size. Both activities described for dextransucrase (dextran formation and acceptor reaction) achieved synthesis whilst the hydrolytic activity of dextranase regulated the product molecular size and acceptor availability. Depending on the reaction conditions, the product oligosaccharide mixtures contained mainly sugars (up to 36%) with degrees of polymerization (DP) varying between 10 and 60 together with lower concentrations of both lower and higher molecular weight sugars. Alterations in substrate and dextranase concentrations (50-400 mg ml(-1) and 2.5-46 U ml(-1), respectively) affected the molecular weight of IMO, the reaction rate and the formation of leucrose. This permitted manipulation of the product characteristics. It was found that higher substrate and dextranase concentrations gave rise to products with lower molecular sizes and a dextransucrase:dextranase ratio of 1: 1 or 1:2 appeared to produce a polymer with a molecular weight which is desirable for prebiotic use. (C) 2004 Elsevier Inc. All rights reserved.

Nanofiltration (NF) of a model sugar solution and a commercial galacto-oligosaccharide mixture has been studied with a cross-flow filtration unit in an attempt to investigate the significance of the operating parameters and evaluate a possible purification process. The pressure, feed concentration and filtration temperature effects were studied in experiments carried out in full recycle mode of operation. The rejection factors of the sugar components present in the solutions increased with increasing pressure due to increased solvent flux and also compaction of the membranes (e.g. rejection factor increased from 0.10 to 0.42 for fructose), with this effect being greater for the low molecular weight sugars in the solutions. Increasing the total sugar concentration of the feed caused a decrease to the rejections of the sugars (e.g. from 0.58 to 0.43 for glucose). Increasing the filtration temperature caused a decrease to the observed rejections of the low molecular weight sugars of the solutions (e.g. from 0.72 to 0.48 for fructose). Continuous diafiltration (CD) purification using NF-CA-50 membranes (at 25 °C) and DS-5-DL membranes (at 60 °C), gave yield values of 14–18% for the monosaccharide, 59–89% for the disaccharide and 81–98% for the trisaccharide (oligosaccharide), respectively. The study clearly demonstrates the potential of cross-flow nanofiltration in the purification of oligosaccharide from mixtures containing contaminant monosaccharides.