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首页> 外文期刊>Biotechnology Progress >Enhanced Heterologous Protein Production in Pichia pastoris Under Increased Air Pressure
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Enhanced Heterologous Protein Production in Pichia pastoris Under Increased Air Pressure

机译：气压增加下巴斯德毕赤酵母中异源蛋白质的产生增强

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摘要

Pichia pastoris is a widely used host for the production of heterologous proteins. In this case, high cell densities are needed and oxygen is a major limiting factor. The increased air pressure could be used to improve the oxygen solubility in the medium and to reach the high oxygen demand of methanol metabolism. In this study, two P. pastoris strains producing two different recombinant proteins, one intracellular (β-galactosidase) and other extracellular (frutalin), were used to investigate the effect of increased air pressure on yeast growth in glycerol and heterologous protein production, using the methanol AOX1-inducible system. Experiments were carried out in a stainless steel bioreactor under total air pressure of 1 bar and 5 bar. The use of an air pressure raise of up to 5 bar proved to be applicable for P. pastoris cultivation. Moreover, no effects on the kinetic growth parameters and methanol utilization (Mut) phenotype of strains were found, while an increase in recombinant β-galactosidase-specific activity (ninefold) and recombinant frutalin production was observed. Furthermore, the air pressure raise led to a reduction in the secreted protease specific activity. This work shows for the first time that the application of an air pressure of 5 bar may be used as a strategy to decrease protease secretion and improve recombinant protein production in P. pastoris.
机译：巴斯德毕赤酵母是广泛用于产生异源蛋白质的宿主。在这种情况下，需要高细胞密度，并且氧气是主要的限制因素。增加的气压可用于改善介质中的氧溶解度，并达到甲醇代谢的高氧需求。在这项研究中，使用两种产生两种不同重组蛋白的巴斯德毕赤酵母菌株，一种是胞内（β-半乳糖苷酶），另一种是胞外（frutalin），用于研究气压升高对甘油中酵母菌生长和异源蛋白质生产的影响。甲醇AOX1诱导系统。实验是在不锈钢生物反应器中于1 bar和5 bar的总气压下进行的。事实证明，使用高达5 bar的气压升高可用于巴斯德毕赤酵母培养。此外，未发现对菌株的动力学生长参数和甲醇利用（Mut）表型的影响，而观察到重组β-半乳糖苷酶比活性（九倍）和重组果胶蛋白产量的增加。此外，气压升高导致分泌的蛋白酶比活性降低。这项工作首次表明，施加5 bar的气压可用作减少蛋白酶分泌和提高巴斯德毕赤酵母中重组蛋白产量的策略。

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