Enhancement of alkaline polygalacturonate lyase production in recombinant Pichia pastoris according to the ratio of methanol to cell concentration |
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Authors: | Wang Yun Wang Zhihao Du Guocheng Hua Zhaozhe Liu Liming Li Jianghua Chen Jian |
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Institution: | Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China. |
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Abstract: | 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. |
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