欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (11): 34-38.doi: 10.11924/j.issn.1000-6850.2012-0774

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

棘孢木霉几丁质酶基因的原核表达、复性、纯化以及酶学性质研究

丛大鹏 李雅华 咸洪泉   

  • 收稿日期:2012-03-07 修回日期:2012-03-13 出版日期:2012-04-15 发布日期:2012-04-15
  • 基金资助:

    山东省自然科学基金

Expression in Escherichia coli, Purification, Renaturation and Characterization of Chitinase Gene from Trichoderma asperelluma

  • Received:2012-03-07 Revised:2012-03-13 Online:2012-04-15 Published:2012-04-15

PDF (PC)

964

摘要：

为高效表达几丁质酶基因tachi1，研究Tachi1的酶学性质。采用PCR技术从棘孢木霉中克隆了几丁质酶基因tachi1并与pEHisTEV原核表达载体融合，转化大肠杆菌BL21。经异丙基-β-D-硫代半乳糖苷(IPTG)诱导，获得了以包涵体形式高效表达的Tachi1。包涵体经洗涤、溶解、复性和纯化后获得了高纯度的Tachi1。复性后的Tachi1具有较高的几丁质酶活性，该几丁质酶反应的最适温度为50℃，最适pH 5.5。几丁质酶在30~35℃下稳定，在pH 3~7之间几丁质酶有较高活性。

关键词: 过硫酸钾, 过硫酸钾

Abstract:

The tachi1 gene of Trichoderma chitinase by Trichoderma asperelluma with pEHisTEV prokaryotic expression vector fusion was highly expressed in E. coli BL21 strains. After induced by isopropyl-β-D-thiogalactoside (IPTG), the recombinant protein Tachi1 was highly expressed in the for mofinclusion bodies. The protein Tachi1 in the inclusion bodies could be solubilized, renatured and purified by a serial of treatments, including washing, denaturing and renaturing as well as purification. The renaturation protein possessed chitinase activity.

中图分类号: