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Heterologous protein production using the Pichia pastoris expression system

机译:使用巴斯德毕赤酵母表达系统生产异源蛋白

摘要

The Pichia pastoris expression system is being used successfully for the production of various recombinant heterologous proteins. Recent developments with respect to the Pichia expression system have had an impact on not only the expression levels that can be achieved, but also the bioactivity of various heterologous proteins. We review here some of these recent developments, as well as strategies for reducing proteolytic degradation of the expressed recombinant protein at cultivation, cellular and protein levels. The problems associated with post-translational modifications performed on recombinant proteins by P. pastoris are discussed, including the effects on bioactivity and function of these proteins, and some engineering strategies for minimizing unwanted glycosylations. We pay particular attention to the importance of optimizing the physicochemical environment for efficient and maximal recombinant protein production in bioreactors and the role of process control in optimizing protein production is reviewed. Finally, future aspects of the use of the P. pastoris expression system are discussed with regard to the production of complex membrane proteins, such as G protein-coupled receptors, and the industrial and clinical importance of these proteins.
机译:巴斯德毕赤酵母表达系统已成功用于各种重组异源蛋白的生产。毕赤酵母表达系统的最新发展不仅影响可达到的表达水平,而且还影响各种异源蛋白的生物活性。我们在这里回顾了这些最新进展,以及在培养,细胞和蛋白质水平上减少表达的重组蛋白的蛋白水解降解的策略。讨论了与巴斯德毕赤酵母对重组蛋白进行的翻译后修饰相关的问题,包括对这些蛋白的生物活性和功能的影响,以及一些使不想要的糖基化最小化的工程策略。我们特别关注优化物理化学环境以在生物反应器中高效,最大程度地重组蛋白质生产的重要性,并综述了过程控制在优化蛋白质生产中的作用。最后,讨论了巴斯德毕赤酵母表达系统使用的未来方面,涉及复杂膜蛋白(例如G蛋白偶联受体)的生产以及这些蛋白的工业和临床重要性。

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