Development of wheat bran hydrolysate as Komagataella phaffii medium for heterologous protein production.

Abstract

Developing a Komagataella phaffii (K. phaffii, named as Pichia pastoris formerly) medium using wheat bran hydrolysate (WBH) is a potential approach for wheat bran utilization and heterologous protein by K. phaffii because K. phaffii is used as protein producer by researchers and engineers widely. In this research, the detoxification process of WBH was optimized to obtain the final procedure as pH adjusting to 10 by calcium hydroxide addition, then, 2.0g/L active carbon absorption followed by 1h shaking and 3,600 × g centrifugation for 10min, finally, 3.75mmol/L sodium thiosulfate addition for 10min shaking followed by 3,600 × g centrifugation for 10min. Recombinant K. phaffii-xynB harboring xylanase B gene from Aspergillus niger ATCC 1015 under alcohol oxidase 1 promoter (PAOX1) was cultivated in detoxified WBH expressing 1059.8U/mL xylanase B which was 90.9% of that in complex medium from Pichia protocols. These researches built a solid base for detoxified WBH as a low-cost medium of K. phaffii to express heterologous protein, also provided a bright outlet for wheat bran utilization.