Biochemical properties and substrate recognition mechanism of GH31 alpha-glucosidase from Bacillus sp AHU 2001 with broad substrate specificity

Saburi, Wataru; Okuyama, Masayuki; Kumagai, Yuya; Kimura, Atsuo; Mori, Haruhide

Biochemical properties and substrate recognition mechanism of GH31 alpha-glucosidase from Bacillus sp AHU 2001 with broad substrate specificity

Saburi, Wataru; Okuyama, Masayuki; Kumagai, Yuya et al.

Biochimie, 2015, 108, 140-148

Permalink : https://hdl.handle.net/2115/58508

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アクセス権	open access
URI	https://hdl.handle.net/2115/58508
タイトル	Biochemical properties and substrate recognition mechanism of GH31 alpha-glucosidase from Bacillus sp AHU 2001 with broad substrate specificity
著者	Saburi, Wataru ; Okuyama, Masayuki ; Kumagai, Yuya ; Kimura, Atsuo ; Mori, Haruhide
e-Rad_Researcher 00598089 著者名 Saburi, Wataru
e-Rad_Researcher 00344490 著者名 Okuyama, Masayuki
e-Rad_Researcher 00589997 著者名 Kumagai, Yuya
e-Rad_Researcher 90186312 著者名 Kimura, Atsuo
e-Rad_Researcher 80241363 著者名 Mori, Haruhide
言語	英語
キーワード	alpha-Glucosidase
	Glycoside hydrolase family 31
	Substrate specificity
	Neotrehalose
	Transglucosylation
発行日	2015-01
出版者	Elsevier
収録物名	Biochimie
巻(号)	108
ページ	140-148
抄録	alpha-Glucosidases are ubiquitous enzymes that hydrolyze the alpha-glucosidic linkage at the non-reducing end of substrates. In this study, we characterized an alpha-glucosidase (BspAG31A) belonging to glycoside hydrolase family 31 from Bacillus sp. AHU 2001. Recombinant B5pAG31A, produced in Escherichia colt, had high hydrolytic activity toward maltooligosaccharides, kojibiose, nigerose, and neotrehalose. This is the first report of an alpha-glucosidase with high activity toward neotrehalose. The transglucosylation products, nigerose, kojibiose, isomaltose, and neotrehalose, were generated from 440 mm maltose. Substitution of Tyr268, situated on the beta -> alpha loop 1 of B5pAG31A, with Trp increased hydrolytic activity toward isomaltose. This mutation reduced the hydrolytic activity toward maltooligosaccharides more than toward kojibiose, nigerose, and neotrehalose. Analysis of the Y173A mutant of B5pAG31A showed that Tyr173, situated on the N-terminal domain loop, is associated with the formation of subsite +2. In Y173A, the k(cad)/K-m for maltooligosaccharides slightly decreased with an increasing degree of polymerization compared with wild type. Among the amino acid residues surrounding the substrate binding site, Va1543 and Glu545 of B5pAG31A were different from the corresponding residues of Bacillus thermoamyloliquefaciens alpha-glucosidase II, which has higher activity toward isomaltose than B5pAG31A. The E545G mutation slightly enhanced isomaltase activity without a large reduction of hydrolytic activities toward other substrates. V543A showed 1.8-3.5-fold higher hydrolytic activities toward all substrates other than neotrehalose compared with wild type, although its preference for isomaltose was unchanged. (C) 2014 Elsevier B.V. and Societe francaise de biochimie et biologie Moleculaire (SFBBM). All rights reserved.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0300-9084
EISSN	0300-9084
NCID	AA00564792
関連情報	PMID 25450253
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.biochi.2014.11.010