食品科学 ›› 2018, Vol. 39 ›› Issue (8): 77-82.doi: 10.7506/spkx1002-6630-201808013

• 生物工程 • 上一篇    下一篇

特基拉芽孢杆菌精氨酸酶基因的重组表达及酶学性质

赵齐1,冯志彬2,秦雪1,徐勤省1,任梦云1,潘潇1,张洪霞1,张娟1,*   

  1. (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)
  • 出版日期:2018-04-25 发布日期:2018-04-17
  • 基金资助:
    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)

Expression in Escherichia coli and Characterization of Arginase from Bacillus tequilensis

ZHAO Qi1, FENG Zhibin2, QIN Xue1, XU Qinxing1, REN Mengyun1, PAN Xiao1, ZHANG Hongxia1, ZHANG Juan1,*   

  1. (1. School of Agriculture, Ludong University, Yantai 264025, China; 2. School of Life Sciences, Ludong University, Yantai 264025, China)
  • Online:2018-04-25 Published:2018-04-17

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)PanD37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pH值为10.0,在pH?8.0~10.0及50~60?℃范围内酶活力稳定,Mn2+、Ni2+、Co2+对酶活力有明显的激活作用,粗酶的活力可达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

Abstract: 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.

Key words: L-arginase, Bacillus tequilensis, L-ornithine, enzymatic properties

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