Molecular mechanism for the substrate specificity of Arthrobacter globiformis M6 α-glucosidase CmmB, belonging to glycoside hydrolase family 13 subfamily 30

Saburi, Wataru; Tagami, Takayoshi; Usui, Takuya; Yu, Jian; Ose, Toyoyuki; Yao, Min; Mori, Haruhide

Molecular mechanism for the substrate specificity of Arthrobacter globiformis M6 α-glucosidase CmmB, belonging to glycoside hydrolase family 13 subfamily 30

Saburi, Wataru; Tagami, Takayoshi; Usui, Takuya et al.

Food bioscience, 2024, 61, 104516

Permalink : https://hdl.handle.net/2115/95586

このアイテムのアクセス数:153件（2026-08-24 16:00 集計）

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論文情報

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アクセス権	open access
URI	https://hdl.handle.net/2115/95586
タイトル	Molecular mechanism for the substrate specificity of Arthrobacter globiformis M6 α-glucosidase CmmB, belonging to glycoside hydrolase family 13 subfamily 30
著者	Saburi, Wataru ; Tagami, Takayoshi ; Usui, Takuya ; Yu, Jian ; Ose, Toyoyuki ; Yao, Min ; Mori, Haruhide
e-Rad_Researcher 00598089 著者名 Saburi, Wataru
e-Rad_Researcher 70709849 著者名 Tagami, Takayoshi
著者名 Usui, Takuya
著者名 Yu, Jian
著者名 Ose, Toyoyuki
著者名 Yao, Min
e-Rad_Researcher 80241363 著者名 Mori, Haruhide
言語	英語
キーワード	alpha-glucosidase
	glycoside hydrolase family 13
	alpha-amylase
	maltooligosaccharide
発行日	2024-06-12
出版者	Elsevier
収録物名	Food bioscience
巻(号)	61
ページ	104516
抄録	alpha-Glucosidase hydrolyzes alpha-D-glucosides to produce alpha-D-glucose. Glycoside hydrolase family 13 (GH13) contains alpha-glucosidases together with various amylolytic enzymes. GH13 alpha-glucosidases fall into several distinct subfamilies, and subfamily 30 (GH13_30) includes numerous Actinomycetes alpha-glucosidases. GH13_30 alpha-glucosidase CmmB from Arthrobacter globiformis, specific to maltooligosaccharides, is involved in the intracellular metabolism of cyclobis-(1 -> 6)-alpha-maltosyl. Herein, the function and structure of CmmB were investigated to advance understanding of the structure-function relationship. CmmB showed the highest kcat/Km for maltose of maltooligosaccharides, and kcat/Km drastically decreased with increasing substrate chain-length. The crystal structures of CmmB in complex with a pseudodisaccharide (acarviosin) and pseudotetrasaccharide (acarbose) were determined at resolutions of 1.60 and 1.70 & Aring;, respectively. The overall structure of CmmB was typical of a GH13 alpha-glucosidase. Most of the structure of the beta ->alpha loop 7 of the catalytic (beta/alpha)8-barrel domain was not determined in the acarbose complex, but the C-terminal side of this loop was modeled in the acarviosin complex. For binding to acarviosin, this loop took on a closed conformation, and I360 and R364 on this loop formed subsite +1 together with H224 and W280. Alanine substitution of these residues indicated that R364 was essential for the catalysis through a hydrogen bond with the O6 of the D-glucose residue in subsite +1. The beta ->alpha loop 7 is flexible upon binding to substrates, but I360 and R364 cannot participate in binding to maltooligosaccharides longer than maltose. The loss of interactions with these residues was concluded to result in the low preference for maltotriose and longer maltooligosaccharides.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
権利情報	© 2024. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	2212-4292
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.fbio.2024.104516