Characterization of purified green bell pepper hydroperoxide lyase expressed by Yarrowia lipolytica:: Radicals detection during catalysis

被引：18
作者：
Santiago-Gomez, Mirna P.
Vergely, Catherine
Policar, Clotilde
Nicaud, Jean-Marc
Belin, Jean-Marc
Rochette, Luc
Husson, Florence
机构：
[1] Univ Bourgogne, ENSBANA, INRA, UMR UB 1232,Lab Microbiol, F-21000 Dijon, France
[2] Fac Med & Pharm, Lab Physiopathol & Pharmacol Cardiovasc Expt, F-21000 Dijon, France
[3] Univ Paris 11, Equipe Chim Bioorgan & Bioinorgan, UMR 8124,CNRS, Inst Chim Mat Orsay,UMR 8182, F-91405 Orsay, France
[4] INRA, CNRS, INAPG, UMR 2585,Lab Microbiol & Genet Mol, F-78850 Thiverval Grignon, France
来源：
ENZYME AND MICROBIAL TECHNOLOGY | 2007年 / 41卷 / 1-2期
关键词：
Yarrowia lipolytica; hydroperoxide lyase; cloning; purification; radicals; EPR spectroscopy;
D O I：
10.1016/j.enzmictec.2006.11.017
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：
The optimization of the expression of recombinant 6-His-tagged HPO lyase in Yarrowia lipolytica is described: 1800 U/L of culture was detected at 24 h of culture on a medium containing olive oil as the sole carbon source. The enzyme was purified by IMAC and showed an optimal pH at 5.5, an optimal temperature at 20 degrees C and a K-m value of 9 mu M with 13-HPOD substrate. The participation of radicals during the catalysis of purified bell pepper fruit hydfoperoxide lyase has been observed by electron paramagnetic resonance spectroscopy and the yet unidentified radical species might be an alkyl or alkoxyl radical linked to the enzyme. (c) 2006 Elsevier Inc. All rights reserved.
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页码：13 / 18
页数：6
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