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Coenzyme precursor-assisted expression of a cholesterol oxidase from Brevibacterium sp. in Escherichia coli

Biotechnology Letters volume 29, pages 761–766 (2007)Cite this article

Abstract

The gene (choB b ), encoding cholesterol oxidase from Brevibacterium sp. CCTCC M201008, was cloned and sequenced by PCR (GenBank accession number: DQ345780). The gene consists of 1653 base pairs and encodes a protein of 551 amino acids. ChoB b exhibited a homology of 98% with cholesterol oxidase gene from Brevibacterium sterolicum ATCC 21387. The cholesterol oxidase gene, cloned in the vector pET-28a, was over-expressed in Escherichia coli BL21–CodonPlus (DE3)-RP grown at 23°C in Luria-Bertani medium containing 50 μM riboflavin, the precursor of the FAD coenzyme of the enzyme. A maximum activity of 3.7 U/mg was obtained from cell free extract of E. coli BL21-CodonPlus (DE3)-RP harboring the pET-28a-choBb.

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  1. Key Laboratory of Industrial Biotechnology of Ministry of Education and School of Biotechnology, Southern Yangtze University, Wuxi, 214036, P.R. China

    Longgang Wang & Wu Wang

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  1. Longgang Wang
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  2. Wu Wang
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Correspondence to Longgang Wang.

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Wang, L., Wang, W. Coenzyme precursor-assisted expression of a cholesterol oxidase from Brevibacterium sp. in Escherichia coli . Biotechnol Lett 29, 761–766 (2007). https://doi.org/10.1007/s10529-006-9295-0

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  • DOI: https://doi.org/10.1007/s10529-006-9295-0

Keywords

  • Brevibacterium sp.
  • Cholesterol oxidase
  • Escherichia coli
  • Over-expression
  • Riboflavin