指状青霉草酰乙酸水解酶的结构特征与原核表达分析  

Analysis of Structural Characteristics and Prokaryotic Expression of Oxaloacetate Hydrolase from Penicillium digitatum

作　　者：胡紫微 李至敏[2] 张梦洁 徐依婷 李志敏[1] HU Zi-wei;LI Zhi-min;ZHANG Meng-jie;XU Yi-ting;LI Zhi-min(College of Bioscience and Bioengineering,Jiangxi Agricultural University,Nanchang 330045,China;College of Science,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]江西农业大学生物科学与工程学院,江西南昌330045 [2]江西农业大学理学院,江西南昌330045

出　　处：《江西农业学报》2021年第5期31-37,共7页Acta Agriculturae Jiangxi

基　　金：江西省自然科学基金项目(20192BAB204011);江西省教育厅科技计划项目(GJJ190202)。

摘　　要：利用生物信息学技术手段分析了柑橘绿霉病病原菌指状青霉(Penicillium digitatum)中草酰乙酸水解酶(Pd OAH)的二级和三级结构,并利用分子生物学技术对重组的Pd OAH进行了原核表达分析和分离纯化。生物信息学分析结果表明Pd OAH的建模结构的质量较高,与其它来源的草酰乙酸水解酶的氨基酸序列具有高度的同源性,且在活性中心氨基酸残基高度保守。重组Pd OAH蛋白在大肠杆菌中进行原核表达,在16℃、0.2 mmol/L IPTG浓度条件下具有较高的表达量,并且有一定的可溶性。利用亲和层析技术分离纯化方法获得了纯度较高的重组Pd OAH蛋白。The secondary and tertiary structures of oxaloacetate hydrolase from Penicillium digitatum(Pd OAH)were analyzed by bioinformatics techniques,and the prokaryotic expression of the recombinant Pd OAH was analyzed and purified by molecular biology techniques.Bioinformatics analysis showed that the quality of modeling structure of Pd OAH was high.Pd OAH was highly homologous to the amino acid sequences of oxaloacetate hydrolases from other sources,and the amino acid residues in the active centers were highly conserved.The recombinant Pd OAH protein induced expression in Escherichia coli,and the expression level and solubility of the Pd OAH protein induced at 16℃with 0.2 mmol/L IPTG concentration were better.The recombinant Pd OAH protein with high purity was purified to be homogenous by affinity chromatography.

关 键 词：指状青霉 草酰乙酸水解酶 生物信息学分析 原核表达 蛋白纯化 

分 类 号：Q556[生物学—生物化学]