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Xylanase B from Clostridium cellulovorans 743B: overexpression, purification, crystallization and X‐ray diffraction analysis
Author(s) -
Nakajima Daichi,
Nagano Akihiko,
Shibata Toshiyuki,
Tanaka Reiji,
Kuroda Kouichi,
Ueda Mitsuyoshi,
Miyake Hideo
Publication year - 2018
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x18000341
Subject(s) - xylanase , cellulosome , chemistry , glycoside hydrolase , cellulase , carbohydrate binding module , biochemistry , crystallography , gene , hydrolysis , enzyme , clostridium thermocellum
Clostridium cellulovorans produces multi‐enzyme complexes called cellulosomes capable of efficiently degrading cellulosic biomass. There are three xylanase genes containing a sequence corresponding to a dockerin domain that are necessary for constructing cellulosomes in the genome. Among the xylanases encoded by these genes, xylanase B (XynB) contains a catalytic domain belonging to glycoside hydrolase family 10 and a carbohydrate‐binding module (CBM) at the N‐terminus, making it a member of CBM family 22. In this study, XynB was cloned, overexpressed, purified and crystallized. XynB was crystallized using the hanging‐drop vapour‐diffusion method in the presence of 0.2 M sodium acetate trihydrate, 0.1 M Tris–HCl pH 8.5, 32%( w / v ) PEG 4000 at 293 K. X‐ray diffraction analysis revealed that the crystal diffracted to 1.95 Å resolution and belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 74.28, b = 77.55, c = 88.20 Å, α = β = γ = 90°. The data‐evaluation statistics revealed high quality of the collected data, thereby establishing a solid basis for determination of the structure of cellulosomal xylanase from C. cellulovorans .