特基拉芽孢杆菌精氨酸酶基因的重组表达及酶学性质  

Expression in Escherichia coli and Characterization of Arginase from Bacillus tequilensis

作　　者：赵齐 冯志彬[2] 秦雪 徐勤省 任梦云 潘潇 张洪霞 张娟 ZHAO Qi;FENG Zhibin;QIN Xue;XU Qinxing;REN Mengyun;PAN Xiao;ZHANG Hongxia;ZHANG Juan(School of Agriculture,Ludong University,Yantai 264025,China;School of Life Sciences,Ludong University,Yantai 264025,China)

机构地区：[1]鲁东大学农学院,山东烟台264025 [2]鲁东大学生命科学学院,山东烟台264025

出　　处：《食品科学》2018年第8期77-82,共6页Food Science

基　　金：农业部转基因生物新品种培育重大专项(2016ZX08004002);烟台市科技计划项目(2015ZH071)

摘　　要：精氨酸酶是一种能够催化精氨酸生成尿素和L-鸟氨酸的碱性水解酶。利用分子克隆的方法得到特基拉芽孢杆菌(Bacillus tequilensis)Pan D37的精氨酸酶基因,在大肠杆菌中异源表达该基因后分析重组酶的性质。结果表明:该精氨酸酶编码297个氨基酸序列;酶的最适反应温度为45℃,最适反应p H值为10.0,在pH 8.0～10.0及50～60℃范围内酶活力稳定,Mn^(2+)、Ni~(^(2+))、Co^(2+)对酶活力有明显的激活作用,粗酶的活力可达1109.8U/m L。Arginase is an alkaline hydrolases that catalyzes the production of urea and L-ornithine from arginine.An arginase gene encoding 297 amino acids was cloned from Bacillus tequilensis PanD37.By recombinant DNA technique,a genetically engineered strain of Escherichia coli was constructed for heterologous expression of the gene.Enzymatic assays indicated that the recombinant enzyme exhibited maximum activity at pH 10.0 and 45℃.Its activity was stable in the temperature range of 50–60℃and in the pH range of 8.0–10.0.The enzymatic activity could be significantly activated by Mn2+,Ni2+,and Co2+.The activity of crude enzyme was 1 109.8 U/mL.

关 键 词：精氨酸酶 特基拉芽孢杆菌 L-鸟氨酸 酶学特性 

分 类 号：Q812[生物学—生物工程]