中文摘要：

透明质酸酶（Hyalurondiase,HAase）是一种具有医药作用的水解酶,为了提高HAase在重组毕赤酵母中的分泌表达量,对重组毕赤酵母诱导阶段的温度进行了优化。研究发现,降低诱导温度可以提高HAase的产量,其中两阶段控制温度诱导策略效果最为显著。诱导阶段0~60 h温度控制在25℃,60~96 h温度控制在22℃,HAase酶活最高为1.18×106U/m L,是诱导温度为30℃时（4.53×105 U/m L）的2.6倍。此外,采用两阶段控制温度诱导策略有利于细胞活性的提高和AOX1启动子的表达。两阶段控制温度诱导策略还可以应用到其他酵母表型中,为毕赤酵母高效表达外源蛋白质提供一种新思路。

英文摘要：

Hyalurondiases（HAase） are a kind of hydrolases with medicine values. In order to improve the expression of HAase in the recombinant Pichia pastoris,the effect of induction temperature on HAase expressed was investigated in the recombinant P. pastoris. The result showed that the production of HAase could be improved with the decrease of induction temperatures,especially,two-stage temperature induction strategy. In this strategy,temperature was controlled at 25 ℃ during the induction stage 0~60 h,and then shifted to 22 ℃ after 60 h. HAase activity reached to 1.18×106 U/mL,2.6 fold improvement compared to induction temperature at 30 ℃（4.53×105 U/mL）. In addition,two-stage temperature induction strategy had an advantage on cell viability and alcohol oxidase activity,which offered a new strategy for expression of exogenous protein in the recombinant P. pastoris.