Optimizing carbon/energy metabolism to enhance monellin production by Pichia pastoris.
- Author:
Qiangqiang HUAI
1
;
Luqiang JIA
1
;
Jian DING
1
;
Shanshan CHEN
1
;
Jiaowen SUN
1
;
Zhongping SHI
1
Author Information
- Publication Type:Journal Article
- Keywords: NADH utilization efficienty; Pichia pastoris; carbon metabolism ratios; induction; monellin
- From: Chinese Journal of Biotechnology 2018;34(2):282-293
- CountryChina
- Language:Chinese
- Abstract: In heterologous protein productions by Pichia pastoris, methanol induction is generally initiated when cell density reaches very high level. However, this traditional strategy suffers with the problems of difficulty in DO control, toxic by-metabolites accumulation and low targeted protein titer. Therefore, initiating methanol induction at lower cell concentration is considered as an alternative strategy to overcome those problems. However, the methanol/energy regulation mechanisms of initiating induction at lower concentration are not clear and seldom reported. In this article, with monellin production as a prototype, we analyzed the methanol/energy metabolism in protein expression process using the strategies of initiating induction at both higher/lower cells concentrations. We attempted to interpret the advantages of the "alternative" strategy, via online measurements of methanol consumption, CO₂ production and O₂ uptake rates. When adopting this "alternative" strategy and maintaining temperature at 30 °C, carbon flux ratio directing into monellin precursors synthesis reached the highest level of 65%. In addition, monellin synthesis was completely associated with cell growth.
