Site-directed mutagenesis of possible catalytic residues of cellobiose 2-epimerase from Ruminococcus albus

Ito, Shigeaki; Hamada, Shigeki; Ito, Hiroyuki; Matsui, Hirokazu; Ozawa, Tadahiro; Taguchi, Hidenori; Ito, Susumu

Site-directed mutagenesis of possible catalytic residues of cellobiose 2-epimerase from Ruminococcus albus

Ito, Shigeaki; Hamada, Shigeki; Ito, Hiroyuki et al.

Biotechnology Letters, 2009, 31(7), 1065-1071

Permalink : https://hdl.handle.net/2115/38754

このアイテムのアクセス数:1,626件（2026-08-08 10:46 集計）

閲覧可能ファイル

ファイル	フォーマット	サイズ	閲覧回数	説明
31-7_p1065-1071	pdf	387 KB	2,143

論文情報

ファイル出力 EndNote Basic出力 Mendeley出力

アクセス権	open access
URI	https://hdl.handle.net/2115/38754
タイトル	Site-directed mutagenesis of possible catalytic residues of cellobiose 2-epimerase from Ruminococcus albus
著者	Ito, Shigeaki ; Hamada, Shigeki ; Ito, Hiroyuki ; Matsui, Hirokazu ; Ozawa, Tadahiro ; Taguchi, Hidenori ; Ito, Susumu
著者名 Ito, Shigeaki
著者名 Hamada, Shigeki
著者名 Ito, Hiroyuki
e-Rad_Researcher 90109504 著者名 Matsui, Hirokazu
著者名 Ozawa, Tadahiro
著者名 Taguchi, Hidenori
著者名 Ito, Susumu
言語	英語
キーワード	Cellobiose 2-epimerase
	Ruminococcus albus
	Homology modeling
	Site-directed mutagenesis
	Epilactose
	Prebiotics
発行日	2009-07
出版者	Springer Netherlands
収録物名	Biotechnology Letters
巻(号)	31(7)
ページ	1065-1071
抄録	Cellobiose 2-epimerase (EC 5.1.3.11) from Ruminococcus albus (RaCE) catalyzes the reversible epimerization of cellobiose and lactose to 4-O-β-D-glucopyranosyl-D-mannose and 4-O-β-D-galactopyranosyl-D-mannose (epilactose). Based on the structure-based amino acid sequence alignment with N-acetyl-D-glucosamine 2-epimerases (EC 5.1.3.8) from porcine kidney and Anabaena sp. CH1 and on the computer-aided model building of the tertiary structure of RaCE, we performed site-directed mutagenesis of possible catalytic residues in the enzyme, and the mutants were expressed in Escherichia coli cells. It was found that R52, H243, E246, W249, M251, W304, E308, H374, and M378 were absolutely required for the activity of RaCE. F114 and W303 (and possibly R377) also contributed to catalysis. These possible catalytic protruded into or near the active-site cleft surrounded by the inner α-helices in the predicted (α/α)6 core barrel structure of RaCE.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	The original publication is available at springerlink.com
PISSN	0141-5492
関連情報	PMID 19330485
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1007/s10529-009-9979-3