Türkanoğlu Özçelik Aysun, Yılmaz Semiramis, Inan Mehmet
Food Safety and Agricultural Research Center, Akdeniz University, Antalya, Turkey.
Department of Food Engineering, Akdeniz University, Antalya, Turkey.
Methods Mol Biol. 2019;1923:97-112. doi: 10.1007/978-1-4939-9024-5_3.
The methylotrophic yeast Pichia pastoris (Komagataella phaffii) is used as an expression system for recombinant protein production for a variety of applications. It grows rapidly on inexpensive media containing methanol, glucose, glycerol, or ethanol as a sole carbon source. P. pastoris makes many posttranslational modifications and produces recombinant proteins either intracellularly or extracellularly. Because of these properties, P. pastoris has become a highly preferred host organism for biotechnology, pharmaceutical industry, and researchers.Recombinant protein production is usually performed under the control of the promoter of the alcohol oxidase gene I (AOX1). The AOX1 promoter is induced by methanol and repressed by glucose and ethanol. The regulation mechanisms of the AOX1 promoter have been studied in recent years. Another promoter used in recombinant protein production is derived from glyceraldehyde 3-phosphate dehydrogenase (GAP). It is a constitutive promoter. Recent literature showed that newly identified promoters of P. pastoris are promising as well, in addition to pAOX1 and pGAP.In this chapter, the regulation mechanisms of inducible pAOX1 and constitutive pGAP promoters are discussed. In addition, here we present an overview about the novel ADH3 promoter and alternative promoters of P. pastoris.
甲基营养型酵母毕赤酵母(Komagataella phaffii)被用作重组蛋白生产的表达系统,用于多种应用。它能在含有甲醇、葡萄糖、甘油或乙醇作为唯一碳源的廉价培养基上快速生长。毕赤酵母能进行许多翻译后修饰,并在细胞内或细胞外产生重组蛋白。由于这些特性,毕赤酵母已成为生物技术、制药行业和研究人员高度青睐的宿主生物。
重组蛋白的生产通常在醇氧化酶基因I(AOX1)启动子的控制下进行。AOX1启动子由甲醇诱导,被葡萄糖和乙醇抑制。近年来,对AOX1启动子的调控机制进行了研究。另一个用于重组蛋白生产的启动子来源于3-磷酸甘油醛脱氢酶(GAP)。它是一个组成型启动子。最近的文献表明,除了pAOX1和pGAP之外,毕赤酵母新发现的启动子也很有前景。
在本章中,将讨论诱导型pAOX1和组成型pGAP启动子的调控机制。此外,我们还将概述毕赤酵母的新型ADH3启动子和其他启动子。