Expression of recombinant enhanced green fluorescent protein provides insight into foreign gene-expression differences between Mut+ and MutS strains of Pichia pastoris.

摘要: Pichia pastoris is a very popular yeast for recombinant protein production, mainly due to the strong, methanol-inducible PAOX1 promoter. Methanol induction however poses several drawbacks. One approach improve processes use MutS strains with reduced methanol catabolic ability. Various reports claim that allows higher production levels than Mut+ but scarcely elaborate on reasons differences. In this study, enhanced green fluorescent was used as -driven reporter investigation of expression differences between and strains. exhibited responses methanol, faster growth (0.07 vs. 0.01 hr-1 ) consumption (2.25 1.81 mmol/gDCW .hr) oxygen (2.2 0.66 mmol/gDCW MutS. yielded more biomass (2.3 1.3 gDCW /L), carbon dioxide analysis bioreactor off-gas suggested considerable amounts were consumed by via dissimilatory pathway. contrast, it demonstrated population switched an induced state rapidly Mut+. addition, 3.4-fold fluorescence per cell (77,509 23,783 SFU) indicative production. The findings verified similar results obtained during lipase. Based in response versus proposed energy availability occurs synthesis, contrary uses maintain high pathways