生物技术进展 ›› 2017, Vol. 7 ›› Issue (3): 253-257.DOI: 10.19586/j.2095-2341.2017.0005

• 研究论文 • 上一篇    

黄豆豆渣中脲酶的提取精制及其影响因素研究

张铁军1,施圆圆1,孔令漪2,曹蓝霄2,周嘉青3   

  1. 1.广州医科大学生命科学学院, 广州 511436;
    2.广州医科大学第三临床学院, 广州 511436;
    3.广州医科大学南山学院, 广州 511436
  • 收稿日期:2017-01-24 出版日期:2017-05-25 发布日期:2017-02-21
  • 作者简介:张铁军,讲师,博士,主要从事生物化学与分子生物学的研究。E-mail:zhang_tiejun@gzhmu.edu.cn
  • 基金资助:
    国家自然科学基金项目(31501169);广东省自然科学基金项目(2016A030310274);广州市属高校科研项目(1201430264);广州医科大学校级科研项目(2013C10)资助。

Study on the Extraction and Factors Affecting Enzyme Activity of Urease from Soybean Dregs

ZHANG Tiejun, SHI Yuanyuan, KONG Lingyi, CAO Lanxiao, ZHOU Jiaqing   

  1. 1.GMU-GIBH Joint School of Life Sciense, Guangzhou Medical University, Guangzhou 511436, China;
    2.The Third Clinical School, Guangzhou Medical University, Guangzhou 511436, China;
    3.Nanshan School, Guangzhou Medical University, Guangzhou 511436, China
  • Received:2017-01-24 Online:2017-05-25 Published:2017-02-21

摘要: 为拓展脲酶的国产化途径,对黄豆豆渣中脲酶的提取进行了研究。建立了从黄豆豆渣中提取脲酶的全套流程,利用盐析法和有机溶剂沉淀法,通过浸提及离心等试验操作手段,将黄豆豆渣中的脲酶进行提取精制,产率为0.1%。同时确定了脲酶的最佳提取条件:其最佳浸提温度为50℃,最佳丙酮浓度为50%,最适pH为7.0。利用纳氏试剂比色法的原理,测得该脲酶在最佳实验条件下的米氏常数Km值为4.11×10-2 mol/L,1 mL脲酶溶液中的酶活力为18.63 U。从黄豆豆渣中提取脲酶,既可解决脲酶的国产化问题,又可提高黄豆豆渣的附加值,研究结果具有良好的工业价值和经济效益。

关键词: 黄豆豆渣, 脲酶, 酶活力, Km, 酶活影响因素

Abstract: The research of extracting urease from soybean dregs was carried out in order to expand the way of urease localization. The research had established a full set of extracting urease from soybean dregs. By use of the method of salting out and organic solvent precipitation, the urease in the soybean dregs was extracted and refined through experiments of extraction and centrifuging, etc., and its yield rate was 0.1%. Meanwhile, the optimum condition of urease extracting was determined: the optimum extracting temperature was 50℃, the optimum acetone concentration was 50%, and the most suitable pH value was 7.0. By the principle of Nessler’s reagent colorimetric method, it was tested that the value of Michaelis constant (Km) was 4.11×10-2 mol/L and the value of enzyme activity in the urease solution of 1 mL was 18.63 U under the optimum conditions. Extracting urease from soybean dregs not only solves the problem of urease localization, but also improves the added value of soybean dregs. The result of the research brings good industrial values and economic benefits.

Key words: soybean dregs, urease, enzyme activity, Km, factors affecting enzyme activity