Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum
Article first published online: 2 SEP 2004
Acta Crystallographica Section D
Volume 60, Issue 9, pages 1665–1667, September 2004
How to Cite
Kumar, R. S., Brannigan, J. A., Pundle, A., Prabhune, A., Dodson, G. G. and Suresh, C. G. (2004), Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum. Acta Crystallographica Section D, 60: 1665–1667. doi: 10.1107/S0907444904017561
- Issue published online: 2 SEP 2004
- Article first published online: 2 SEP 2004
- Received 17 June 2004, accepted 16 July 2004
- conjugated bile salt hydrolase;
- Ntn hydrolase;
- Bifidobacterium longum;
- penicillin V acylase.
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 P622 with unit-cell parameters a = b = 124.86, c = 219.03 Å, and the trigonal space group P321, 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.