Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents

被引：23

作者：

Mathieu, Sandrine

Bigey, Frederic

Procureur, Jerome

Terrier, Nancy

Gunata, Ziya ^{[1
]}

机构：

[1] Univ Montpellier 2, ENSAM, INRA, UMR IR2B, F-34060 Montpellier 1, France

[2] INRA, ENSAM, UMR SPO, Equipe Biol Integrat, F-34060 Montpellier, France

来源：

BIOTECHNOLOGY LETTERS | 2007年 / 29卷 / 05期

关键词：

carotenoid cleavage dioxygenase; C-13-norisoprenoids; spectrophotometric assay; water-miscible solvents;

D O I：

10.1007/s10529-007-9315-8

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A recombinant carotenoid cleavage dioxygenase from Vitis vinifera L. was produced by Escherichia coli as a fusion with the glutathione-S-transferase (GST) protein under different bacterial growth conditions. The enzyme production was monitored by a GST assay. Addition of Triton X-100 prior to bacterial cell disruption doubled the release of soluble protein. A simple spectrophotometric enzyme assay was developed to measure carotenoid cleavage activity using lutein as substrate. Enzyme activity showed a 26-fold increase with the addition of 10% (v/v) acetone in the reaction mixture.

引用

收藏

页码：837 / 841

页数：5

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