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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Ключевые слова АГРОВОК

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

Библиографическая информация

Опубликовано в

Biotechnology letters

Том 29 Выпуск 5 Нумерация страниц 761 - 766 ISSN 0141-5492

Издатель

Kluwer Academic Publishers

Другие темы

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

Язык

Английский

Тип

Journal Article; Text

В АГРИСе с: 2024-02-27

Формат: MODS

Поставщик данных

Эту запись предоставил National Agricultural Library

Откройте коллекцию этого поставщика данных в AGRIS

Ссылки

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

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