摘要
以毕赤酵母的醇氧化酶(AOX)为指标,以玻璃珠破碎法为对象,建立一种简单易行的毕赤酵母细胞的破碎工艺。考察玻璃珠用量、振荡时间、细胞液体积对毕赤酵母释放醇氧化酶的影响,进而利用Box-Behnken设计组合优化这3种因素,以响应面模型和一个2阶多项式方程拟合了这3种影响因素和醇氧化酶比酶活的关系。结果表明:玻璃珠破碎毕赤酵母的最佳条件为0.23 g玻璃珠、1.04 m L细胞液、103 s振荡时间。在最佳破碎条件下,醇氧化酶比酶活为8.86 U/mg。确定了玻璃珠破碎毕赤酵母细胞的条件,而且操作简便,可以同时处理多个样品,为重组毕赤酵母的筛选提供了有效方法。
In order to lyze the Pichia pastoris to obtain alcohol oxidase,we optimized the lysis process assisted by glass beads including the paraimeters of glass beads,vertex time,and cell volume,by single factor test and response surface methodology. Box-Behnken design was done with three levels of these three factors using specific alcohol oxidase activity as response. The relationship between specific alcohol oxidase activity and the three factors was a second-order polynomial equation. This model was analyzed by ANOVA,and validated by experiment. The optimized P. pastoris lysis condition was vortex time 103 s,glass beads 0. 23 g,cell volume 1.04 m L,with the results of 8.86 U / mg protein specific alcohol oxidase activity. This work provides an easy lysis procedure for P. pastoris.
出处
《生物加工过程》
CAS
2016年第1期30-35,共6页
Chinese Journal of Bioprocess Engineering
基金
国家自然科学基金(21306112)
上海市自然科学基金(13ZR1429100)
上海高校青年教师培养资助计划(slg14037)
教育部留学回国人员科研启动基金
关键词
毕赤酵母
玻璃珠
破碎
醇氧化酶
响应面优化
Pichia pastoris
glass beads
lysis
alcohol oxidase
response surface methodology
