Open AccessCrystallization and preliminary crystallographic analysis of β‐mannanase from Bacillus licheniformis
Author(s) -
Songsiriritthigul Chomphunuch,
Lapboonrueng Sasithorn,
Roytrakul Sittiruk,
Haltrich Dietmar,
Yamabhai Montarop
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110049067
Subject(s) - bacillus licheniformis , mannan , crystallization , monoclinic crystal system , chemistry , polyethylene glycol , lysozyme , crystallography , escherichia coli , nuclear chemistry , peg ratio , chromatography , crystal structure , biochemistry , polysaccharide , biology , bacteria , organic chemistry , genetics , bacillus subtilis , finance , gene , economics
The mannan endo‐1,4‐β‐mannosidase (ManB) from Bacillus licheniformis strain DSM13 was overexpressed in Escherichia coli . Purification of the thermostable and alkali‐stable recombinant mannanase yielded approximately 50 mg enzyme per litre of culture. Crystals were grown by hanging‐drop vapour diffusion using a precipitant solution consisting of 12%( w / v ) PEG 8000, 0.2 M magnesium acetate tetrahydrate and 0.1 M MES pH 6.5. The protein crystallized in the monoclinic space group P 2 1 , with two molecules per asymmetric unit and unit‐cell parameters a = 48.58, b = 91.75, c = 89.55 Å, β = 98.29°, and showed diffraction to 2.3 Å resolution.