Identification of Bacillus selenitireducens MLS10 maltose phosphorylase possessing synthetic ability for branched α-d-glucosyl trisaccharides

• Published in: Carbohydrate research Vol. 360; pp. 25 - 30
• Main Authors: Nihira, Takanori, Saito, Yuka, Kitaoka, Motomitsu, Otsubo, Ken’ichi, Nakai, Hiroyuki
• Format: Journal Article
• Language: English
• Published: OXFORD Elsevier Ltd 01.10.2012; Elsevier
• Subjects: Bacillus - enzymology; Bacillus selenitireducens; Biocatalysis; Biochemistry & Molecular Biology; Branched α-d-glucosyl trisaccharide; Carbohydrate Conformation; Chemistry; Chemistry, Applied; Chemistry, Organic; Cloning, Molecular; Disaccharides - biosynthesis; Disaccharides - chemistry; glucose; Glucosyltransferases - genetics; Glucosyltransferases - isolation & purification; Glucosyltransferases - metabolism; Glycoside hydrolase family 65; glycosides; Hydrogen-Ion Concentration; Kinetics; Life Sciences & Biomedicine; maltose; Maltose - analogs & derivatives; Maltose - biosynthesis; Maltose - chemistry; Maltose phosphorylase; mannose; phosphorylase; Physical Sciences; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Reverse phosphorolysis; Science & Technology; Substrate Specificity; Temperature; trisaccharides; Trisaccharides - biosynthesis; Trisaccharides - chemistry; Branched α-d-glucosyl trisaccharide; Maltose phosphorylase; Glycoside hydrolase family 65; Reverse phosphorolysis; STEAROTHERMOPHILUS SK-1; OLIGOSACCHARIDES; ENZYMATIC-SYNTHESIS; MICROBIOTA; Branched alpha-D-glucosyl trisaccharide; CLOSTRIDIUM-PHYTOFERMENTANS; LACTOBACILLUS-ACIDOPHILUS NCFM; NUCLEOTIDE; GENE; PURIFICATION; TREHALOSE PHOSPHORYLASE
• ISSN: 0008-6215; 1873-426X
• DOI: 10.1016/j.carres.2012.07.014

[Display omitted] ► A unique maltose phosphorylase utilizing glucobioses as acceptor was identified. ► The structures of products were identified as branched α-d-glucosyl trisaccharides. ► The enzyme was suggested to possess a binding site for C2 substituent of d-glucose. We discovered an inverting maltose phosphorylase (Bsel2056) belonging to glycoside hydrolase family 65 from Bacillus selenitireducens MLS10, which possesses synthetic ability for α-d-glucosyl disaccharides and trisaccharides through the reverse phosphorolysis with β-d-glucose 1-phosphate as the donor. Bsel2056 showed the flexibility for monosaccharide acceptors with alternative C2 substituent (2-amino-2-deoxy-d-glucose, 2-deoxy-d-arabino-hexose, 2-acetamido-2-deoxy-d-glucose, d-mannose), resulting in production of 1,4-α-d-glucosyl disaccharides with strict regioselectivity. In addition, Bsel2056 synthesized two maltose derivatives possessing additional d-glucosyl residue bound to C2 position of the d-glucose residue at the reducing end, 1,4-α-d-glucopyranosyl-[1,2-α-d-glucopyranosyl]-d-glucose and 1,4-α-d-glucopyranosyl-[1,2-β-d-glucopyranosyl]-d-glucose, from 1,2-α-d-glucopyranosyl-d-glucose (kojibiose) and 1,2-β-d-glucopyranosyl-d-glucose (sophorose), respectively, as the acceptors. These results suggested that Bsel2056 possessed a binding space to accommodate the bulky C2 substituent of d-glucose.