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Crystal structure of an acetylesterase from Talaromyces cellulolyticus and the importance of a disulfide bond near the active site
Author(s) -
Watanabe Masahiro,
Fukada Harumi,
Inoue Hiroyuki,
Ishikawa Kazuhiko
Publication year - 2015
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2015.03.020
Subject(s) - catalytic triad , active site , chemistry , esterase , enzyme , hydrolysis , hydrolase , stereochemistry , mutagenesis , acylation , site directed mutagenesis , biochemistry , catalysis , mutation , mutant , gene
Carbohydrate esterase catalyzes the de‐ O or de‐ N ‐acylation of substituted saccharides in plant cell walls and thus has great potential for industrial biomass saccharification. We recently identified the putative carbohydrate esterase family 3 (CE3) from Talaromyces cellulolyticus . Here, we prepared the recombinant catalytic domain of the enzyme and crystallized it. The crystal structure was determined to 1.5 Å resolution. From the structural analysis, it was elucidated that a n‐octyl‐β‐ d ‐glucopyranoside bound to near the catalytic triad (Ser10, Asp179 and His182) and was buried in the active site cavity. Site‐directed mutagenesis showed that the N‐terminal disulfide bond located near the catalytic triad is involved in the activity and structural stability of the enzyme.