Synthesis of ¥-Galactooligosaccharide Using Bifidobacterial ¥-Galactosidase Purified from Recombinant Escherichia coli

Oh, Seung Keun; Youn, So Youn; Park, Myung Soo; Kim, Hyoung‐Geun; Baek, Nam‐In; Li, Zhipeng; Ji, Geun Eog

doi:10.4014/jmb.1702.02058

Cited by 20 publications

(21 citation statements)

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“…This is obvious, especially when comparing its growth and fermentation activity scores with 4′GOS-P, which is also free of monosaccharides. These results substantiate the idea that oligosaccharides produced with enzymes from probiotic strains can have a more pronounced effect on the growth of these probiotic strains [16,17,18,43].…”

Section: Discussionsupporting

confidence: 85%

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Fermentability of a Novel Galacto-Oligosaccharide Mixture by Lactobacillus spp. and Bifidobacterium spp.

et al. 2018

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This study aimed to investigate the specific growth stimulation of certain desired intestinal bacteria by a novel galacto-oligosaccharide mixture, which was produced with a β-galactosidase from a potential probiotic Lactobacillus isolate that contained mainly oligosaccharides of β-1,3 and β-1,6 glycosidic linkages (termed Lb-GOS) using single-strain fermentations. The composition of this Lb-GOS mixture was 33.5% disaccharides, 60.5% trisaccharides, 4.8% tetrasaccharides, and 1.0% pentasaccharides with a negligible amount of monosaccharides, lactose, and lactobionic acid (0.3%). Eight Lactobacillus spp. strains and three Bifidobacterium spp. strains were used in single-strain fermentations to determine the fermentation activity scores of this Lb-GOS preparation compared to two commercially available prebiotic mixtures, 4′GOS-P and Vivinal GOS (V-GOS). The highest scores were obtained when L. reuteri Lb46 and the two Bifidobacterium strains, B. animalis subsp. lactis Bif1 and Bif3, were grown on these galacto-oligosaccharide mixtures. In addition, the Lb-GOS mixture was found to have higher fermentation activity scores; hence, it stimulated the growth of these probiotic strains more than 4′GOS-P and V-GOS, which may be attributed to the different glycosidic linkage types that are found in the Lb-GOS mixture compared to the other two commercial preparations. These findings suggested that the Lb-GOS mixture that is described in this work should be of interest for the formulations of new carbohydrate-based functional food ingredients.

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Section: Discussionsupporting

confidence: 85%

“…These oligosaccharides might then be preferentially utilized by the producing probiotics, as was shown for GOS produced by Bifidobacterium spp. derived β-galactosidases [16,17,18].…”

Section: Introductionmentioning

confidence: 99%

Fermentability of a Novel Galacto-Oligosaccharide Mixture by Lactobacillus spp. and Bifidobacterium spp.

et al. 2018

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“…BgaC exhibited transglycosylation activity at a relatively low lactose concentration (~200 mM) compared to known and commercial β-galactosidases that require high lactose concentrations (40% w/v or 1.17 M) (Guerrero et al 2015;Hsu et al 2007;Oh et al 2017;Reuter et al 1999). The ability of this enzyme to synthesise transglycosylation products, presumably GOS, could result in reduced GOS production costs using BgaC compared to currently available commercial enzymes.…”

Section: Discussionmentioning

confidence: 98%

“…Some unrelated Bifidobacterium βgalactosidases have been isolated and characterised through expression of the recombinant enzyme in E . c o l i , a nd am on g st t he s e, a fe w d is pl ay e d transglycosylation activity and synthesis of GOS (Arreola et al 2014;Hsu et al 2007;Oh et al 2017;Yi et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Metagenomic identification, purification and characterisation of the Bifidobacterium adolescentis BgaC β-galactosidase

Mulualem

Agbavwe

Ogilvie

et al. 2021

Appl Microbiol Biotechnol

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Members of the human gut microbiota use glycoside hydrolase (GH) enzymes, such as β-galactosidases, to forage on host mucin glycans and dietary fibres. A human faecal metagenomic fosmid library was constructed and functionally screened to identify novel β-galactosidases. Out of the 16,000 clones screened, 30 β-galactosidase-positive clones were identified. The β-galactosidase gene found in the majority of the clones was BAD_1582 from Bifidobacterium adolescentis, subsequently named bgaC. This gene was cloned with a hexahistidine tag, expressed in Escherichia coli and His-tagged-BgaC was purified using Ni2+-NTA affinity chromatography and size filtration. The enzyme had optimal activity at pH 7.0 and 37 °C, with a wide range of pH (4–10) and temperature (0–40 °C) stability. It required a divalent metal ion co-factor; maximum activity was detected with Mg2+, while Cu2+ and Mn2+ were inhibitory. Kinetic parameters were determined using ortho-nitrophenyl-β-d-galactopyranoside (ONPG) and lactose substrates. BgaC had a Vmax of 107 μmol/min/mg and a Km of 2.5 mM for ONPG and a Vmax of 22 μmol/min/mg and a Km of 3.7 mM for lactose. It exhibited low product inhibition by galactose with a Ki of 116 mM and high tolerance for glucose (66% activity retained in presence of 700 mM glucose). In addition, BgaC possessed transglycosylation activity to produce galactooligosaccharides (GOS) from lactose, as determined by TLC and HPLC analysis. The enzymatic characteristics of B. adolescentis BgaC make it an ideal candidate for dairy industry applications and prebiotic manufacture.Key points• Bifidobacterium adolescentis BgaC β-galactosidase was selected from human faecal metagenome.• BgaC possesses sought-after properties for biotechnology, e.g. low product inhibition.• BgaC has transglycosylation activity producing prebiotic oligosaccharides.

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“…35 Other Table 4 Increase in the cell density between time 0 and time 24 h and final pH for pure cultures of selected probiotic strains grown on various oligosaccharide substrates not purified and purified studies have evaluated the prebiotic effect of lactose and lactulose derived oligosaccharides on pure cultures of potential probiotic strains by assessing growth and/or metabolite production. 31,35,40,47,48 However, substrate consumption has not been assessed. Our results show that, although no strong growth was obtained with purified substrates, the microorganisms were able to metabolize to different extents the oligosaccharides derived from lactose and lactulose, in agreement with previous studies using this type of bacteria and oligosaccharides.…”

Section: Determination Of the Prebiotic Capacity Of Crl450-gos And Crmentioning

confidence: 99%

Prebiotic galactooligosaccharides production from lactose and lactulose by Lactobacillus delbrueckii subsp. bulgaricus CRL450

Fara

Sabater²,

Palacios

et al. 2020

Food Funct.

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Synthesis of ¥-Galactooligosaccharide Using Bifidobacterial ¥-Galactosidase Purified from Recombinant Escherichia coli

Cited by 20 publications

References 0 publications

Fermentability of a Novel Galacto-Oligosaccharide Mixture by Lactobacillus spp. and Bifidobacterium spp.

Fermentability of a Novel Galacto-Oligosaccharide Mixture by Lactobacillus spp. and Bifidobacterium spp.

Metagenomic identification, purification and characterisation of the Bifidobacterium adolescentis BgaC β-galactosidase

Prebiotic galactooligosaccharides production from lactose and lactulose by Lactobacillus delbrueckii subsp. bulgaricus CRL450

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