食品与发酵工业 >

2020 , Vol. 46 >Issue 24: 58 - 65

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.024535

研究报告

源于解淀粉芽孢杆菌酸性木聚糖酶酶学性质的研究

  • 郑亚伦 ,
  • 夏瑛 ,
  • 李良 ,
  • 董孝元 ,
  • 方尚玲 ,
  • 陈茂彬 ,
  • 李琴
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  • 1(湖北工业大学 生物工程与食品学院,湖北 武汉,430070)
    2(黄鹤楼酒业有限公司,湖北 武汉,430050)
    3(武汉雅仕博科技有限公司,湖北 武汉,430061)
    4(湖北省酿造工艺与装备工程技术中心,湖北 武汉,430070)
硕士研究生(李琴讲师为通讯作者,E-mail:17888820100@163.com)

收稿日期: 2020-05-24

  修回日期: 2020-09-02

  网络出版日期: 2021-01-13

基金资助

国家自然科学基金(31901634)

收起

Enzymatic properties of acid resistant xylanase from Bacillus amyloliquefaciens

  • ZHENG Yalun ,
  • XIA Ying ,
  • LI Liang ,
  • DONG Xiaoyuan ,
  • FANG Shangling ,
  • CHEN Maobin ,
  • LI Qin
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  • 1(School of food and Biological engineering, Hubei University of Technology, Wuhan 430070, China)
    2(Huanghelou Liquor Company Limited, Wuhan 430050, China)
    3(Wuhan yashibo Technology Company Limited, Wuhan 430061, China)
    4(Center for Brewing Technology & Equipment Research, Hubei University of Technology, Wuhan 430070, China)

Received date: 2020-05-24

  Revised date: 2020-09-02

  Online published: 2021-01-13

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摘要

该文从解淀粉芽孢杆菌中克隆得到木聚糖酶基因,在大肠杆菌中实现高效率的异源性表达,设计响应面实验获得最佳的表达优化条件,得到更高酶活力的产物。利用异丙基-β-D-硫代吡喃半乳糖苷(isopropyl-β-D-thiogalactoside, IPTG)溶液对重组菌株BA-TB-1进行诱导表达,对诱导条件进行优化。实验发现该木聚糖酶对酸具有较好的耐受性,在pH值为5.0~6.0时有较高的酶活性。在37 ℃, pH5.0和pH 5.0~7.0条件下分别培养1 h,测定酶活力,残余酶活性分别保持在68%和50%以上。最后通过响应面法得到最佳的优化条件,在此最优条件下进行诱导表达。预测值和实际值分别为550.139、548.87 U/mL,预测值与实际值相近,说明应用响应面法优化木聚糖酶的表达条件可行,该实验结果为大量生产酸性木聚糖酶奠定了一定的理论基础。

关键词： 木聚糖酶; 解淀粉芽孢杆菌; 耐酸性; 酶学性质; 克隆表达

本文引用格式

郑亚伦 , 夏瑛 , 李良 , 董孝元 , 方尚玲 , 陈茂彬 , 李琴 . 源于解淀粉芽孢杆菌酸性木聚糖酶酶学性质的研究[J]. 食品与发酵工业, 2020 , 46(24) : 58 -65 . DOI: 10.13995/j.cnki.11-1802/ts.024535

Abstract

Xylanase is a type of hydrolytic enzyme that can catalyze the hydrolysis of xylan in plant hemicellulose, it is widely used in various industries and the market now has higher requirements for its production and activity. A xylanase gene was cloned from Bacillus amyloliquefaciens and heterogenous expressed in Escherichia coli. Isopropyl-β-D-thiogalactoside (IPTG) was adopted to induce the expression of recombinate strain BA-TB-1 and response surface experiments were designed to optimize the expression conditions. The predicted activity and experimental activity were 550.139 and 548.87 U/mL, respectively, indicating that it was feasible to apply response surface method to optimize the expression conditions of xylanase. Further results showed that xylanase had relatively high enzyme activity at pH 5.0 to 6.0, proving a good acid-resistant property. Incubated at 37℃, pH 5.0 and pH 5.0-7.0 for 1 h, the residual enzyme activity remained above 68% and 50%, respectively. The results lay some foundation for the mass production of acidic xylanase.

Key words： xylanase; Bacillus amyloliquefaciens; acid resistance; enzymatic properties; cloning and expression

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