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《Biotechnology Bulletin》 2008-03
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Expressions of Endoglucanase Ⅱ Gene(egⅡ)in Escherichia coli and Pichia pastoris

Wang Jin1 Wu Minchen2 Zhou Chenyan1(1School of Biotechnology,Jiangnan University Key Laboratory of Industry Biotechnology Under Ministry of Education,Jiangnan University,Wuxi 214122;2School of Medicine and Pharmaceutics,Jiangnan University,Wuxi 214122)  
The mature peptide of egⅡ(1 194bp)was amplified with a template of the recombinant plasmid pTG19-T-egⅡ and specific primers designed according to the published cDNA secqunce of egⅡ from Trichoderma viride WL 0422.It was inserted into vectors of pET-28a(+)and pPIC9K,which were transformed to Escherichia coli Rosset(DE3)and Pichia pastoris GS115,respectively.Product of expression in E.coli was 15% of bacterial whole protein,and did not have any activity of endoglucanase.With methanol induction,the product of expression in P.pastoris had an activity of endoglucanase,and the expression level exceeded 2 788U/ml in shake culture.The optimum pH and temperature of the enzyme secreted by P.pastoris was 4.5 and 50℃.The enzyme was quite stable between pH 3.5 to 6.0,and the relative activities were remained over 80%.It was also stable at the temperature below 50℃,and the relative activities were remained over 90%.
【Fund】: 国家自然科学基金(20776061)
【CateGory Index】: Q78
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