Enhancement of alkaline polygalacturonate lyase production in recombinant Pichia pastoris according to the ratio of methanol to cell concentration
2009
Wang, Yun | Wang, Zhihao | Du, Guocheng | Hua, Zhaozhe | Liu, Liming | Li, Jianghua | Chen, Jian
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 140gL⁻¹ after 19h 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 0.163-0.171gg⁻¹, and PGL activity and productivity reached 430UmL⁻¹ and 4.34UmL⁻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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