| 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 |
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| 引用本文： | 丛大鹏,李雅华,咸洪泉.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[J].中国农学通报,2012,28(11):34-38. |
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| 作者姓名： | 丛大鹏 李雅华 咸洪泉 |
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| 作者单位： | 青岛农业大学生命科学学院,山东青岛,266109 |
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| 基金项目： | 山东省自然科学基金“生防木霉菌的分子标记技术研究”（ZR2010CM040）; 青岛农业大学高层次人才启动基金“木霉菌几丁质酶新基因克隆表达及酶学性质分析”（631108） |
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| 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pH5.5。几丁质酶在30～35℃下稳定,在pH3～7之间几丁质酶有较高活性。
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| 关 键 词： | 棘孢木霉 几丁质酶基因 表达 原核表达 蛋白质纯化 酶学性质 |
| 收稿时间： | 3/7/2012 12:00:00 AM |
| 修稿时间： | 2012/3/13 0:00:00 |
Expression in Escherichia coli, Purification, Renaturation and Characterization of Chitinase Gene from Trichoderma asperelluma |
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| Cong Dapeng
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Li Yahua
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Xian Hongquan.Expression in Escherichia coli, Purification, Renaturation and Characterization of Chitinase Gene from Trichoderma asperelluma[J].Chinese Agricultural Science Bulletin,2012,28(11):34-38. |
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| Authors: | Cong Dapeng Li Yahua Xian Hongquan |
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| Institution: | (Qingdao Agricultural University,Life Sciences Institute,Qingdao Shandong 266109) |
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| 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. |
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| Keywords: | Trichoderma asperelluma Chitinase gene expression protein purification renaturation characterization |
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