Wang Yun, Wang Zhihao, Du Guocheng, Hua Zhaozhe, Liu Liming, Li Jianghua, Chen Jian
Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China.
Bioresour Technol. 2009 Feb;100(3):1343-9. doi: 10.1016/j.biortech.2008.07.049. Epub 2008 Sep 11.
Polygalacturonate lyase (PGL) production by Pichia pastoris GS115 was used as a model to study the mechanism and strategy for enhancing heterologous protein production. It was found that the ratio of methanol to cell concentration had a significant influence on PGL production. In this study, an advanced glycerol exponential feeding strategy was developed for biomass accumulation in cell growth phase, by which cell concentration reached 140 g L(-1) after 19 h glycerol feeding. In subsequent production phase, a methanol feeding profile was proposed according to the optimal ratio of methanol to cell concentration at a range of 0.163-0.171 g g(-1), and PGL activity and productivity reached 430 U mL(-1) and 4.34 U mL(-1)h(-1), respectively. The strategy for enhancing PGL production by controlling the optimal ratio may provide an alternative approach to enhance heterologous protein production with P. pastoris.
以毕赤酵母GS115生产聚半乳糖醛酸裂解酶(PGL)为模型,研究提高异源蛋白产量的机制和策略。发现甲醇与细胞浓度的比例对PGL产量有显著影响。在本研究中,开发了一种先进的甘油指数补料策略用于细胞生长阶段的生物量积累,通过该策略,在补料甘油19小时后细胞浓度达到140 g L(-1)。在随后的生产阶段,根据甲醇与细胞浓度的最佳比例(范围为0.163 - 0.171 g g(-1))提出了甲醇补料曲线,PGL活性和生产率分别达到430 U mL(-1)和4.34 U mL(-1)h(-1)。通过控制最佳比例提高PGL产量的策略可能为利用毕赤酵母提高异源蛋白产量提供一种替代方法。