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Protein Expr Purif. 2006 Jun;47(2):607-13. Epub 2006 Jan 30.

Cloning, overexpression, purification, and characterization of O-acetylserine sulfhydrylase-B from Escherichia coli.

Author information

1
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, Japan.

Abstract

O-Acetylserine sulfhydrylase-B (OASS-B, EC 2.5.1.47) is one of the two isozymes produced by Escherichia coli that catalyze the synthesis of L-cysteine from O-acetyl-L-serine and sulfide. The cysM gene encoding OASS-B was cloned and the enzyme was overexpressed in E. coli using pUC19 with a lacUV5 promoter. The enzyme was purified to homogeneity, as evidenced by SDS-PAGE. Approximately 300 mg of purified OASS-B was obtained from 1600 mL of culture broth with a purification yield of 60% or higher. The purified OASS-B was characterized and its properties compared with OASS-A. OASS-B did not form a complex with E. coli serine acetyltransferase (SAT, EC 2.3.1.30) and showed a wide range of substrate specificity in nonproteinaceous amino acid synthesis.

PMID:
16546401
DOI:
10.1016/j.pep.2006.01.002
[Indexed for MEDLINE]

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