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中国生物工程杂志

China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (6): 45-51    DOI:
    
Prokaryotic Expression of tachi2 Gene from Trichoderma asperellum and Characterization of Recombinant Enzyme
ZHANG Jun-xia, CONG Da-peng, LI Ya-hua, XIAN Hong-quan
College of Life Science, Qingdao Agricultural University, Qingdao 266109, China
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Abstract  

Objective: To realize high level prokaryotic expression of the tachi2 gene from Trichoderma asperellum and characterize the recombinant enzyme. Methods: The tachi2 gene was amplified by PCR, and recombinant plasmid pEHISTEV /tachi2 was constructed successfully by inserting the tachi2 gene into pEHISTEV. The recombinant plasmid pEHISTEV/tachi2 was transformed into Escherichia coli BL21, which was induced with isopropyl-β-D-thiogalactoside (IPTG). The recombinant protein Tachi2 was purified and renatured to analyze enzymatic properties. Results:The recombinant protein Tachi2 was expressed successfully in the form of inclusion body in the recombinant E.coli. The molecular weight of Tachi2 was about 44 kDa, Tachi2 was purified and renatured by a serial of treatments, the renatured protein had high chitinase activity. The optimum temperature and pH for the enzyme activity were 40℃ and 7.0 respectively. The enzyme activity was stable under 40℃ and in the pH range of 6~9, its activity was significantly reduced by 0.05 mol/L of Cu2+ and Zn2+. Conclusion:The recombinant strain Escherichia coli BL21 showed high level prokaryotic expression of the tachi2 gene. And the enzymatic properties were further defined. It will provide theoretical basis for application and further research in this chitinase.



Key wordsTrichoderma asperellum      Chitinase      Prokaryotic expression      Enzymatic properties     
Received: 01 April 2013      Published: 25 June 2013
ZTFLH:  Q93  
Cite this article:

ZHANG Jun-xia, CONG Da-peng, LI Ya-hua, XIAN Hong-quan. Prokaryotic Expression of tachi2 Gene from Trichoderma asperellum and Characterization of Recombinant Enzyme. China Biotechnology, 2013, 33(6): 45-51.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2013/V33/I6/45

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