Expression, refolding and characterization of human brain serine racemase in Escherichia coli with N-terminal His-tag.
Human brain serine racemase (hSR) was expressed in large amounts in E. coli with N-terminal His-tag (His-hSR). His-hSR expressed in inclusion body was solubilized and purified to homogeneity by Ni-NTA affinity column. Purified His-hSR was refolded in Tween 20/cycloamylose with approximately 50% efficiency, and refolded His-hSR was isolated by Q Sepharose column chromatography. The refolding conditions are described in detail. His-hSR catalyzed the elimination of L-Ser as well as L-Ser-O-sulfate to form pyruvate.
Nagayoshi
Chizuru
C
Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
Ishibashi
Matsujiro
M
Kita
Yoshiko
Y
Matsuoka
Masaaki
M
Nishimoto
Ikuo
I
Tokunaga
Masao
M
D000595:Amino Acid Sequence; D001921:Brain / Q000201:enzymology; D002847:Chromatography, Agarose; D003902:Detergents; D004926:Escherichia coli; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D006801:Humans; D002479:Inclusion Bodies; D008969:Molecular Sequence Data; D017510:Protein Folding; D019998:Racemases and Epimerases / Q000096:biosynthesis*; D011994:Recombinant Proteins; D012995:Solubility; D000099300:Serine Racemase
D003902:Detergents; D019998:Racemases and Epimerases; D011994:Recombinant Proteins; D000099300:Serine Racemase