Wurm David Johannes, Spadiut Oliver
Institute of Chemical, Environmental and Bioscience Engineering, Research Division Biochemical Engineering, Vienna University of Technology (TU Wien), Vienna, Austria.
Methods Mol Biol. 2019;1923:323-333. doi: 10.1007/978-1-4939-9024-5_15.
Pichia pastoris is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. A prominent promoter system for recombinant protein production in P. pastoris is the promoter of alcohol oxidase (P) which is induced by methanol, but repressed by several other carbon sources, like glucose and glycerol. Thus, typical cultivation strategies for such P. pastoris strains describe two different phases: growth on a carbon source, like glycerol, to get a high biomass concentration, followed by the induction of recombinant protein production by methanol. However, cells barely grow on methanol resulting in only moderate productivity in such bioprocesses. To enhance productivity, it is common to employ mixed substrate feeding strategies. The knowledge of certain strain-specific parameters is required to be able to set up such mixed feed fed-batch cultivations to avoid methanol accumulation and guarantee highest productivity. Here, we present an efficient strategy comprising only one experiment to determine the settings of such a mixed feed system based on the physiology of the respective yeast strain.
毕赤酵母是工业生物技术中重组生产蛋白质最重要的宿主生物之一。毕赤酵母中用于重组蛋白生产的一个重要启动子系统是甲醇诱导的乙醇氧化酶启动子（P），但它会被其他几种碳源（如葡萄糖和甘油）抑制。因此，此类毕赤酵母菌株的典型培养策略描述了两个不同阶段：在碳源（如甘油）上生长以获得高生物量浓度，随后通过甲醇诱导重组蛋白生产。然而，细胞在甲醇上几乎不生长，导致此类生物过程中的生产力仅为中等水平。为了提高生产力，通常采用混合底物补料策略。为了能够建立这种混合补料分批培养以避免甲醇积累并保证最高生产力，需要了解某些菌株特异性参数。在此，我们提出一种仅包含一个实验的有效策略，以根据相应酵母菌株的生理学来确定这种混合补料系统的设置。
