摘要
原动蛋白2(PK2)是近年发现的一个具有多种生物学功能的蛋白质因子。采用Design-Expert 7.0.0软件对重组PK2表达的最优诱导条件进行响应面分析。结果表明,诱导起始时机与诱导物浓度是影响重组PK2表达水平的显著性因子。同时,诱导时间-诱导时机、诱导时机-诱导物浓度之间的相互作用对PK2的表达水平具有显著性影响。试验数据拟合与极值分析表明最优诱导条件为:工程菌OD600为0.60时,采用终浓度为0.62 mmol/L的IPTG在37℃诱导培养2.82 h,此时重组PK2的表达水平为170.73 mg/L。优化后的诱导条件导致重组PK2的表达水平提高了51.1%,而所需IPTG减少38.0%,诱导时间缩短53.0%,有利于大量制备重组PK2进行后续功能研究及应用研究。
Prokineticin 2 （ PK2 ） has been recently identified as a multi-functional factor involved in a variety of physiological processes, including gastrointestinal mobility, circadian rhythms, angiogenesis and neurogenesis. Herein, response surface methodology （ RSM ） was applied to optimize induction conditions for high level expression of PK2 by recombinant Escherichia coli BL21 （ DE3 ）. It was found that induction- starting cell density and inducer concentration had significant effects on production of PK2. In addition, the interaction between post-induction time and time-point of induction and that between induction-starting cell density and inducer concentration were significant. The optimum levels of these three variables were as follows ： 2.82 h for post-induction time, OD600 0.60 for induction-starting cell density and 0.62 mmol/L of IPTG for inducer concentration. A maximum 170.73 mg/L of recombinant PK2 was attained in the validation experiment under optimized condition, which was 51.1% higher than that before optimization. The results will be helpful for large-scale preparation of PK2 for further function and application study.
出处
《生物技术通报》
CAS
CSCD
北大核心
2012年第8期175-180,共6页
Biotechnology Bulletin
基金
江苏大学高级人才启动基金资助项目(1281370001)
关键词
响应面法
原动蛋白2(PK2)
诱导时间
诱导物浓度
诱导时机
优化
Response surface methodology （ RSM ）
Pmkineticin 2 （ PK2 ）
Post-induction time
Inducer concentration
Time-pointof induction
Optimization
