期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 117, 期 5, 页码 1394-1405出版社
WILEY
DOI: 10.1002/bit.27303
关键词
bioprocess; growth decoupled; Komagataella phaffii; methanol induction; Mut(-)
资金
- Austrian Science Fund [W1224]
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 performing, producing 531 mg/L HSA and 1631 mg/L vHH. 61% of the HSA was produced in the methanol only phase where no biomass growth was observed, representing a special case of growth independent production. By using the Mut(-) phenotype, the oxygen demand, heat output, and specific methanol uptake (q(methanol)) in the methanol phase were reduced by more than 80% compared with the Mut(S) phenotype. The highlighted improved process parameters coupled with growth independent protein production are overlooked benefits of the Mut(-) strain for current and future applications in the field of recombinant protein production.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据