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Optimisation of expression and immobilized metal ion affinity chromatographic purification of recombinant (HiS)6-tagged cytochrome P450 hydroperoxide lyase in Escherichia coli

被引：21
作者：
Delcarte, J
Fauconnier, ML
Jacques, P
Matsui, K
Thonart, P
Marlier, M
机构：
[1] Agr Res Ctr, B-5030 Gembloux, Belgium
[2] Agr Univ Gembloux, Unit Gen & Organ Chem, B-5030 Gembloux, Belgium
[3] Agr Univ Gembloux, Walloon Ctr Ind Biol, B-5030 Gembloux, Belgium
[4] Yamaguchi Univ, Fac Agr, Dept Biol Chem, Yamaguchi 7538515, Japan
来源：
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2003年 / 786卷 / 1-2期
关键词：
expression; purification; Escherichia coli; hydroperoxide lyase; cytochrome P450; histidine tag;
D O I：
10.1016/S1570-0232(02)00815-2
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Fatty acid hydroperoxide lyase (HPL) is a cytochrome P450 acting on fatty acid's hydroperoxides in many plants. The optimisation of the expression of recombinant (His)(6)-tagged HPL in Escherichia coli is described: the highest HPL production yield were obtained with TB medium supplemented with 2.5 mM delta-aminolevulinic acid and 0.5 mM IPTG. For the first time, the time course expression of a plant P450 in a bench-scale fermentor is detailed and the amount of recombinant HPL production is 16.3 mg/l. The UV-Visible spectrum of the recombinant (HiS)(6)-tagged HPL have been recorded after a Ni2+-based affinity chromatography (IMAC). (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码：229 / 236
页数：8
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