Open AccessSubstrate recognition by a bifunctional GH30‐7 xylanase B from Talaromyces cellulolyticus
Author(s) -
Nakamichi Yusuke,
Watanabe Masahiro,
Matsushika Akinori,
Inoue Hiroyuki
Publication year - 2020
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12873
Subject(s) - xylobiose , chemistry , xylanase , glycoside hydrolase , hydrolase , pichia pastoris , stereochemistry , enzyme , xylose , residue (chemistry) , glycosyl , biochemistry , bifunctional , phosphofructokinase 2 , substrate (aquarium) , subfamily , fermentation , biology , recombinant dna , ecology , gene , catalysis
Xylanase B, a member of subfamily 7 of the GH30 (glycoside hydrolase family 30) from Talaromyces cellulolyticus ( Tc Xyn30B), is a bifunctional enzyme with glucuronoxylanase and xylobiohydrolase activities. In the present study, crystal structures of the native enzyme and the enzyme–product complex of Tc Xyn30B expressed in Pichia pastoris were determined at resolutions of 1.60 and 1.65 Å, respectively. The enzyme complexed with 2 2 ‐(4‐ O ‐methyl‐α‐ d ‐glucuronyl)‐xylobiose (U 4m2 X) revealed that Tc Xyn30B strictly recognizes both the C‐6 carboxyl group and the 4‐ O ‐methyl group of the 4‐ O ‐methyl‐α‐ d ‐glucuronyl side chain by the conserved residues in GH30‐7 endoxylanases. The crystal structure and site‐directed mutagenesis indicated that Asn‐93 on the β2‐α2‐loop interacts with the non‐reducing end of the xylose residue at subsite‐2 and is likely to be involved in xylobiohydrolase activity. These findings provide structural insight into the mechanisms of substrate recognition of GH30‐7 glucuronoxylanase and xylobiohydrolase.