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

Wang, Longgang; Wang, Wu

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

2007

Wang, Longgang | Wang, Wu

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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AGROVOC Keywords

amino acids brevibacterium chemistry cholesterol electrophoresis enzymology escherichia coli gene expression genes genetics pharmacology polymerase chain reaction protein riboflavin riboflavin sequence analysis temperature

Bibliographic information

Published in

Biotechnology letters

Volume 29 Issue 5 Pagination 761 - 766 ISSN 0141-5492

Publisher

Kluwer Academic Publishers

Other Subjects

Drug effects; Brevibacterium sterolicum; Polyacrylamide gel; Cholesterol oxidase; Amino acid sequence; Sequence homology; Amino acid; Molecular sequence data; Gene overexpression

Language

English

Type

Journal Article; Text

In AGRIS since: 2024-02-27

Format: MODS

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This bibliographic record has been provided by National Agricultural Library

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Links

DOI http://dx.doi.org/10.1007/s10529-006-9295-0

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