耐热羧肽酶Taq基因在毕赤酵母中的表达
Sheng wu gong cheng xue bao = Chinese journal of biotechnology（2014）
Abstract
为了获取表达羧肽酶Taq毕赤酵母工程菌,通过密码子优化,依据毕赤酵母密码子偏爱性,在体外合成了栖热水生菌的耐热羧肽酶Taq基因.将该基因克隆到毕赤酵母表达载体pHBM905A上并引入6×His标签,构建了重组质粒pHBM905A-Cpase Taq.将该重组质粒转化毕赤酵母GS115,经1％甲醇诱导表达72 h,酶产量达0.1 mg/mL.对纯化的重组酶进行酶活性分析表明在75℃,pH为7.5时,该酶比酶活性为80 U/mg.本研究首次证明了羧肽酶Taq能在毕赤酵母中有效分泌表达,可以被大量制备,进而为多肽水解为氨基酸奠定工业基础.
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Key words
thermostable carboxypeptidase taq gene,pichia pastoris gs115
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