Characterization of methanol utilization negative Pichia pastoris for secreted protein production: New cultivation strategies for current and future applications.

ABSTRACT: The methanol utilization (Mut) phenotype in the yeast Pichia pastoris (syn. Komagataella spp.) is defined by the deletion of the genes AOX1 and AOX2. The Mut^- phenotype cannot grow on methanol as a single carbon source. We assessed the Mut^- phenotype for secreted recombinant protein production. The methanol inducible AOX1 promoter (P_AOX1 ) was active in the Mut^- phenotype and showed adequate eGFP fluorescence levels and protein yields (Y_P/X ) in small-scale screenings. Different bioreactor cultivation scenarios with methanol excess concentrations were tested using P_AOX1 HSA and P_AOX1 vHH expression constructs. Scenario B comprising a glucose-methanol phase and a 72-hr-long methanol only phase was the best performi

SUBMITTER: Zavec D 

PROVIDER: S-EPMC7187134 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature