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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Mots clés AGROVOC

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

Informations bibliographiques

Publié dans

Biotechnology letters

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

Editeur

Kluwer Academic Publishers

D'autres materias

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

Langue

anglais

Type

Journal Article; Text

Sur AGRIS depuis: 2024-02-27

Format: MODS

Fournisseur de données

Cette notice bibliographique a été fournie par National Agricultural Library

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Liens

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

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