摘要
按毕赤酵母偏爱密码子,将来自Yersinia rohdei的植酸酶基因(YrAPPA)进行了化学合成,并成功在毕赤酵母GS-115中异源分泌表达,重组植酸酶分泌量能达到72μg/mL。酶学特性研究结果表明:酶比活达到1 500 U/mg,重组植酸酶的最适反应温度和pH分别为55℃和4.5;纯化的重组植酸酶能耐受60℃高温,且pH 3—7.5范围内重组植酸酶有良好的稳定性;Cu^(2+)、Zn^(2+)、Fe^(3+)对植酸酶活性有抑制作用;重组植酸酶有抗胃蛋白酶和胰蛋白酶水解特性。
Yersinia rohdei phytase gene （YrAPPA） was chemically synthesized according to the codon usage of Pichia pastoris and successfully expressed in P. pastoris GS-115, with the secretion yield of recombinant phytase being up to about 72μg/mL. The enzymatic characterization showed that the specific activity of enzyme is 1 500 U/rag, and the optimal reacting temperature and pH of recombinant phytase are respectively 55℃ and 4.5;The purified recombinant phytase can tolerate high temperature at 60℃ and has a good stability within pH 3-7.5 ;The phytase whose activity is inhibited by Cu^2＋, Zn^2＋ and Fe^3＋ has characteristics resistant to trypsin and pepsin.
出处
《上海农业学报》
CSCD
2016年第6期18-25,共8页
Acta Agriculturae Shanghai
基金
上海市科委农业成果转化项目(143919NO300)
上海市种业发展项目[沪农科种字(2016)第1-2号]
关键词
植酸酶
酶学特性
毕赤酵母
电击转化
异源表达
Phytase
Enzymatic property
Pichia pastosis
Electroporation
Heterologous expression
