毕赤酵母
乙醇氧化酶
发酵
酵母
甘油
化学
甲醇
重组DNA
基质(水族馆)
毕赤酵母
补料分批培养
生物化学
生物
基因
有机化学
生态学
作者
Chun Yeon Lee,Atsushi Nakano,Norio Shiomi,Eun Kyu Lee,Shigeo Katoh
标识
DOI:10.1016/s0141-0229(03)00146-7
摘要
Effects of regulated substrate feed on the cell growth and recombinant protein expression by the Pichia system were studied by applying either constant feed rates or variable feed rates in DO-stat fed-batch fermentation. Chimeric α-amylase (Amy1A/3D) that consists of 159 aa of one isozyme (Amy1A) of rice α-amylases followed by 251 aa of another (Amy3D) was used as a model protein. In the glycerol fed-batch (alcohol oxidase I derepression) phase, a step-decreasing glycerol feed control allowed a high growth rate (0.086 h−1) and high cell density (97.2 g DCW l−1). In the induction phase, methanol feed rate had a significant effect on the specific expression rate of Amy1A/3D. A constant feed rate of 1.0 ml l−1 min−1 was found to be optimum for a high specific expression rate of Amy1A/3D (0.091 mg g DCW−1 h−1). Under this condition, allowing a higher specific methanol consumption rate, a maximum concentration of 340 mg l−1 of Amy1A/3D and the expression yield of 2.16 mg Amy1A/3D g-methanol−1 were attained. This result strongly demonstrated that the efficiency of recombinant protein expression was directly dependent on methanol utilization in Pichia system.
科研通智能强力驱动
Strongly Powered by AbleSci AI