Development of an industrial medium and a novel fed-batch strategy for high-level expression of recombinant β-mananase by Pichia pastoris

Abstract

An industrial medium, Corn Steep Liquor Powder Dextrose (CSD medium) was developed for constitutive expression of recombinant β-mananase by Pichia pastoris. The β-mananase activity (513U/mL) with CSD medium was 1.64- and 2.5-fold higher than with YPD and BSM in shaken flasks. The β-mananase productivity with CSD medium was 61.0U/mLh, which was 1.7- and 2.5-fold higher than with YPD and BSM in a 5-L fermenter based on a novel fed-batch strategy combining the real-time exponential feed mode with the DO-stat feed mode. The β-mananase activity, dry cell weight and the recombinant enzyme reached up to 5132U/mL, 110.0g/L and 4.50g/L after 50h cultivation in a 50-L fermenter. The high efficient expression of recombinant β-mananase by P. pastoris indicated that CSD medium and the novel fed-batch strategy have great potential for the production of recombinant β-mananase in industrial fermentation.