Expression, Refolding and Characterization of Human Brain Serine Racemase in Escherichia coli with N-Terminal His-Tag

Title: Expression, Refolding and Characterization of Human Brain Serine Racemase in Escherichia coli with N-Terminal His-Tag

Volume: 12 Issue: 5

Author(s): Chizuru Nagayoshi, Matsujiro Ishibashi, Yoshiko Kita, Masaaki Matsuoka, Ikuo Nishimoto and Masao Tokunaga

Affiliation:

Keywords: human brain serine racemase, d-ser, nmda receptor, refolding, detergents, cycloamylose

Abstract: Human brain serine racemase (hSR) was expressed in large amounts in E. coli with N-terminal His-tag (HishSR). 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 ∼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.

Cite this article as:

Nagayoshi Chizuru, Ishibashi Matsujiro, Kita Yoshiko, Matsuoka Masaaki, Nishimoto Ikuo and Tokunaga Masao, Expression, Refolding and Characterization of Human Brain Serine Racemase in Escherichia coli with N-Terminal His-Tag, Protein & Peptide Letters 2005; 12 (5) . https://dx.doi.org/10.2174/0929866054395284