Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum.
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 A, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 A. The crystals diffracted X-rays to 2.5 A spacing. Structure determination using the multiple isomorphous replacement method is in progress.
Kumar
R Suresh
RS
Division of Biochemical Sciences, National Chemical Laboratory, Pune 411 008, India.
Brannigan
James A
JA
Pundle
Archana
A
Prabhune
Asmita
A
Dodson
Guy G
GG
Suresh
C G
CG
D000581:Amidohydrolases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D001644:Bifidobacterium / Q000201:enzymology* / Q000235:genetics; D003460:Crystallization; D003627:Data Interpretation, Statistical; D008027:Light; D010399:Penicillin Amidase / Q000737:chemistry; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012542:Scattering, Radiation; D014961:X-Ray Diffraction
D000581:Amidohydrolases; D010399:Penicillin Amidase; C023154:choloylglycine hydrolase