Improved enzyme production on corncob hydrolysate by a xylose-evolved Pichia pastoris cell factory

The global shift from the usage of crude oil in bio-production is receiving much attention owing to environmental concern associated with fossil fuel. Lignocellulosic biomass (LB) is a good carbon candidate for bio-production because it is environmental-friendly. Corncob being one of such LB is rich in glucose and xylose, which can be utilized for bio-production. We co-utilize these sugars for the production of enzymes from Pichia pastoris GS115 (Wild Type: WT). Glucose utilization was efficient from synthetic and real hydrolysate but xylose utilization was very low, hence, the need for optimization. Mutants were selected upon Adaptive Laboratory Evolution to efficiently utilize xylose. As expected, all the mutants examined showed improved xylose utilization but surprisingly, there was only 1.8 g/l residual xylose in the 50th generation (GS50). The 30th evolutionary generation (GS30) compared well with the WT by completely utilizing the glucose and also accumulated 48 OD₆₀₀ cell biomass, which is the highest among all the strains evaluated. More importantly, GS30 secreted 72.6 U/ml and 45.1 U/ml β-galactosidase and β-mannanase on hydrolysate respectively, which are higher than the titre for the WT. Conclusively, this study demonstrated the efficacy of corn corncob hydrolysate in biomanufacturing and gives insight for the optimization study.