Brassica napus soluble epoxide hydrolase (BNSEH1) - Cloning and characterization of the recombinant enzyme expressed in Pichia pastoris

文献类型： 外文期刊

第一作者： Bellevik, S

作者： Bellevik, S;Zhang, JM;Meijer, J

作者机构：

关键词： epoxide hydrolase;oilseed rape;yeast;recombinant enzyme;stilbene oxide

期刊名称：EUROPEAN JOURNAL OF BIOCHEMISTRY （ 影响因子：3.579； ）

ISSN： 0014-2956

年卷期： 2002 年 269 卷 21 期

页码：

收录情况： SCI

摘要： Epoxide hydrolase (EC 3. 3.2.3) in plants is involved in the metabolism of epoxy fatty acids and in mediating defence responses. We report the cloning of a full-length epoxide hydrolase cDNA (BNSEH1) from oilseed rape ( Brassica napus) obtained by screening of a cDNA library prepared from methyl jasmonate induced leaf tissue, and the 5'-RACE technique. The cDNA encodes a soluble protein containing 318 amino acid residues. The identity on the protein level is 85% to an Arabidopsis soluble epoxide hydrolase (sEH) and 50-60% to sEHs cloned from other plants. A 5 x His tag was added to the N-terminus of the BNSEH1 and the construct was over-expressed in the yeast Pichia pastoris. The recombinant protein was recovered at high levels after Ni-agarose chromatography of lysed cell extracts, had a molecular mass of 37 kDa on SDS/PAGE and cross-reacted on Western blots with antibodies raised to a sEH from Arabidopsis thaliana. BNSEH1 was shown to be a monomer by gel filtration analysis. The activity was low towards cis stilbene oxide but much higher using trans-stilbene oxide as substrate with V-max of 0.47 mumol.min.mg(-1), K-m of 11 muM and k(cat) of 0.3 s(-1). The optimum temperature of the recombinant enzyme was 55 degreesC and the optimum pH 6-7 for trans-stilbene oxide hydrolysis. The isolation of BNSEH1 will facilitate metabolic engineering of epoxy fatty acid metabolism for functional studies of resistance and seed oil modi cation in this important oilcrop.

分类号：

  • 相关文献

[1]Genetic and biochemical characterization of a protease-resistant mesophilic beta-mannanase from Streptomyces sp S27. Shi, Pengjun,Yuan, Tiezheng,Zhao, Junqi,Huang, Huoqing,Luo, Huiying,Meng, Kun,Wang, Yaru,Yao, Bin.

[2]A novel highly acidic beta-mannanase from the acidophilic fungus Bispora sp MEY-1: gene cloning and overexpression in Pichia pastoris. Luo, Huiying,Wang, Yaru,Wang, Hui,Yang, Jun,Yang, Yuhui,Huang, Huoqing,Yang, Peilong,Bai, Yingguo,Shi, Pengjun,Yao, Bin,Fan, Yunliu.

[3]Cloning, sequencing, and expression of a novel epoxide hydrolase gene from Rhodococcus opacus in Escherichia coli and characterization of enzyme. Liu, Zhiqiang,Li, Yin,Xu, Yingying,Ping, Lifeng,Zheng, Yuguo.

[4]Isolation and identification of a novel Rhodococcus sp ML-0004 producing epoxide hydrolase and optimization of enzyme production. Liu, Zhiqiang,Li, Yin,Ping, Lifeng,Xu, Yingying,Cui, Fengjie,Xue, Yaping,Zheng, Yuguo. 2007

[5]Attraction of Coffee Bean Weevil, Araecerus fasciculatus, to Volatiles from the Industrial Yeast Kluyveromyces lactis. Mei, Xiang-Dong,She, Dongmei,Ning, Jun,Zhang, Xiao-Fang,Li, Yao-Fa,Zhang, Tao.

[6]Effects of dietary live and heat-inactive baker's yeast on growth, gut health, and disease resistance of Nile tilapia under high rearing density. Ran, Chao,Huang, Lu,Hu, Jun,He, Suxu,Li, Zhimin,Wang, Yibing,Liu, Zhi,Xu, Li,Yang, Yalin,Zhou, Zhigang,Tacon, Philippe.

[7]Du1, encoding a novel Prp1 protein, regulates starch biosynthesis through affecting the splicing of Wx(b) supercript stop pre-mRNAs in rice (Oryza sativa L.). Zeng, Dali,Yan, Meixian,Wang, Yonghong,Liu, Xinfang,Qian, Qian,Li, Jiayang.

[8]Identification and characterization of PtDGAT2B, an acyltransferase of the DGAT2 acyl-Coenzyme A: Diacylglycerol acyltransferase family in the diatom Phaeodactylum tricornutum. Gong, Yangmin,Zhang, Junping,Guo, Xiaojing,Wan, Xia,Liang, Zhuo,Hu, Chuan Jiong,Jiang, Mulan,Zhang, Junping.

[9]Characterization of a cold responsive HbICE1 gene from rubber trees. Deng, Xiao Min,Wang, Jian Xiao,Li, Yan,Wang, Jing,Tian, Wei-Min,Deng, Xiao Min,Wang, Jian Xiao,Tian, Wei-Min.

[10]Investigation of Microbial Communities of Chinese Sourdoughs Using Culture-Dependent and DGGE Approaches. Zhang, Guohua,Sadiq, Faizan A.,Zhu, Liying,Liu, Tongjie,Yang, Huanyi,He, Guoqing,Zhang, Guohua,Sadiq, Faizan A.,Zhu, Liying,Liu, Tongjie,Yang, Huanyi,Wang, Xin,He, Guoqing,Zhu, Liying,Wang, Xin.

[11]Microbial Diversity and Biochemical Analysis of Suanzhou: A Traditional Chinese Fermented Cereal Gruel. Qin, Huibin,Sun, Qinghui,Pan, Xuewei,Yang, Hongjiang,Qin, Huibin,Qiao, Zhijun. 2016

[12]Prey diet quality affects predation, oviposition and conversion rate of the predatory mite Neoseiulus barkeri (Acari: Phytoseiidae). Yang, Kang,Wang, Endong,Xu, Xuenong.

[13]Production and Characteristics of High Quality Vinegar from Sugarcane Juice. Lin, Bo,Li, Yang-Rui,Zheng, Feng-Jin,Sun, Jian,Li, Zhi-Chun. 2015

[14]Identification and Characterization of a Glyoxalase I Gene in a Rapeseed Cultivar with Seed Thermotolerance. Yan, Guixin,Lv, Xiaodan,Gao, Guizhen,Li, Feng,Li, Jun,Qiao, Jiangwei,Xu, Kun,Chen, Biyun,Wang, Limin,Xiao, Xin,Wu, Xiaoming. 2016

[15]Preparation and Characteristics of Sugarcane Low Alcoholic Drink by Submerged Alcoholic Fermentation. Lin, Bo,Wang, Tian-Shun,Li, Yang-Rui,Zheng, Feng-Jin. 2013

[16]Generation of High Titre Antibodies to Pokeweed Antiviral Protein (PAP) in Rabbits Using Synthetic Peptides and their Use in Detecting PAP in Transgenic Petunia and Yeast. Li, Yu,Wang, Xifeng. 2012

[17]Characterization of a laccase gene from the white-rot fungi Trametes sp 5930 isolated from Shennongjia Nature Reserve in China and studying on the capability of decolorization of different synthetic dyes. Yang, Yang,Ma, Fuying,Yu, Hongbo,Fan, Fangfang,Zhang, Xiaoyu,Yang, Yang,Wan, Xia,Jiang, Mulan. 2011

[18]Cloning and Functional Characterization of a Fatty Acyl-Acyl Carrier Protein Thioesterase Gene (BnFatB) in Brassica napus L.. Tan, X. L.,Huang, Q.,Tan, R. K.,Zhang, Zh. Y.,Wang, Zh.,Li, X. F.,Wu, L.,Lu, Ch. M.. 2015

[19]Long Chain Acyl-Coenzyme A Synthetase 4 (BnLACS4) Gene from Brassica napus Enhances the Yeast Lipid Contents. Tan Xiao-li,Zheng Xiang-feng,Zhang Zhi-yan,Wang Zheng,Xia Heng-chuan,Gu Shou-lai,Lu Chang-ming. 2014

[20]Effects of different forms of yeast Saccharomyces cerevisiae on growth performance, intestinal development, and systemic immunity in early-weaned piglets. Jiang, Zongyong,Wei, Shaoyong,Wang, Zhilin,Hu, Shenglan,Zheng, Chuntian,Hu, Youjun,Wang, Li,Ma, Xianyong,Yang, Xuefen,Jiang, Zongyong,Zhu, Cui,Chen, Zhuang. 2015

作者其他论文 更多>>

微信小程序