Increased production of alkaline polygalacturonate lyase in the recombinant Pichia pastoris by controlling cell concentration during continuous culture

Recombinant alkaline polygalacturonate lyase (PGL) production by recombinant Pichia pastoris GS115 was selected as a model to study as a continuous culture strategy for enhancing heterologous protein production based on controlling methanol feeding (CCCM culture) or on dual carbon source feeding (CCCD culture). Using the CCCM process with a dry cell weight of 75 g/L regulated by controlling methanol concentration in the induction media, the final PGL activity was 441.9 U/mL. The PGL productivity (Qv) and the average specific enzyme production rate (Qx) were 4.65 U mL−1 h−1 and 84.5 U g−1 h−1, an increase of 42.1% and 191.2%, respectively, over what was achieved with traditional fed-batch culture with high cell density. The control strategies also reduced proteolytic degradation by 84.1% in the fermentation broth and increased cell viability by 12.2%.