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GH36 α-galactosidase from Lactobacillus plantarum WCFS1 synthesize Gal-α-1,6 linked prebiotic α-galactooligosaccharide by transglycosylation.

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Relation http://ir.cftri.com/14393/
https://doi.org/10.1016/j.ijbiomac.2019.12.032
 
Title GH36 α-galactosidase from Lactobacillus plantarum WCFS1 synthesize Gal-α-1,6 linked prebiotic α-galactooligosaccharide by transglycosylation.
 
Creator Deepesh, Panwar
Shubhashini, A.
Sachin, R. Chaudhari
Harish Prashanth, K. V.
Mukesh, Kapoor
 
Subject 07 Enzyme Biochemistry
 
Description α-Galactosidases are potent industrial glycoside hydrolases which are relatively less explored for their transglycosylation potential, especially from Lactobacillus genera. A GH36 α-galactosidase from Lactobacillus plantarum WCFS1 was cloned and over expressed in Hi-control Escherichia coli BL21(DE3). Ni-NTA affinity gel chromatography resulted in purified α-galactosidase (LpαG; specific activity 3077.35 U mg−1) having a monomeric weight of ~80 kDa with 29.3% yield. Size exclusion chromatography of LpαG showed native molecular mass of ~240.5 kDa. LpαG displayed optimum activity at pH 6 and 37 °C. The Km, Vmax and kcat/Km of LpαG towards pNPαGal were found to be 0.93 mM and 714.3 μmol ml−1 min−1 and 12,075 s−1 mM−1, respectively. LpαG displayed maximum transglycosylation activity towards melibiose substrate (as both donor and acceptor) and synthesized majorly a trisaccharide with 0.26 mg ml−1 yield. Nuclear magnetic resonance (NMR) characterization revealed that trisaccharide consist of only single species of α-linked galactooligosaccharide (manninotriose; α-d-Galp-(1 → 6)-α-d-Galp-(1 → 6)-d-Glcp) with α-(1 → 6) regioselectivity. Manninotriose displayed prebiotic property by supporting the growth of probiotic L. plantarum WCFS1 and Bifidobacteria adolescentis DSM 20083.
 
Date 2020
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/14393/1/International%20Journal%20of%20Biological%20Macromolecules%202020.pdf
Deepesh, Panwar and Shubhashini, A. and Sachin, R. Chaudhari and Harish Prashanth, K. V. and Mukesh, Kapoor (2020) GH36 α-galactosidase from Lactobacillus plantarum WCFS1 synthesize Gal-α-1,6 linked prebiotic α-galactooligosaccharide by transglycosylation. International Journal of Biological Macromolecules, 144. pp. 334-342. ISSN 0141-8130