Xylanase B from Clostridium cellulovorans 743B: overexpression, purification, crystallization and X-ray diffraction analysis.

ABSTRACT: 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 P212121, with unit-cell parameters a = 74.28, b = 77.55, c = 88.20?Å, ? = ? = ? = 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.

SUBMITTER: Nakajima D 

PROVIDER: S-EPMC5947682 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature