Overexpression, crystallization, and preliminary X-ray crystallographic analysis of the alanine racemase from Enterococcus faecalis v583.
Alanine racemase, a bacterial enzyme belonging to the fold-type III group of pyridoxal 5'-phosphate (PLP)-dependent enzymes, has been shown to catalyze the interconversion between L- and D-alanine. The alanine racemase from the pathogenic bacterium Enterococcus faecalis v583 has been overexpressed in E. coli and was shown to crystallize an enzyme at 295 K, using polyethylene glycol (PEG) 8000 as a precipitant. X-ray diffraction data to 2.5 A has been collected using synchrotron radiation. The crystal is a member of the orthorhombic space group, C222(1), with unit cell parameter of a=94.634, b=156.516, c=147.878 A, and alpha=beta;=gamma=90 degrees. Two or three monomers are likely to be present in the asymmetric unit, with a corresponding Vm of 3.38 A3 Da(-1) and 2.26 A Da(-1) and a solvent content of 63.7% and 45.5%, respectively.
Priyadarshi
Amit
A
Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul 136-701, Korea.
Lee
Eun Hye
EH
Sung
Min Woo
MW
Kim
Jae-Hee
JH
Ku
Min-Je
MJ
Kim
Eunice Eunkyeong
EE
Hwang
Kwang Yeon
KY
D000411:Alanine Racemase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001709:Biotechnology; D003460:Crystallization; D018360:Crystallography, X-Ray; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000411:Alanine Racemase