Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum
Author(s) -
Kumar R. Suresh,
Brannigan James A.,
Pundle Archana,
Prabhune Asmita,
Dodson Guy G.,
Suresh C. G.
Publication year - 2004
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s0907444904017561
Subject(s) - crystallization , chemistry , crystallography , protein crystallization , conjugated system , hydrolase , hydrolysis , bifidobacterium longum , crystal (programming language) , salt (chemistry) , enzyme , bifidobacterium , biochemistry , organic chemistry , polymer , programming language , fermentation , computer science , lactobacillus
Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging‐drop vapour‐diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P 622 with unit‐cell parameters a = b = 124.86, c = 219.03 Å, and the trigonal space group P 321, with unit‐cell parameters a = b = 125.24, c = 117.03 Å. The crystals diffracted X‐rays to 2.5 Å spacing. Structure determination using the multiple isomorphous replacement method is in progress.