Cregg J M, Vedvick T S, Raschke W C
Department of Chemical and Biological Sciences, Oregon Graduate Institute of Science and Technology, Beaverton 97006-1999.
Biotechnology (N Y). 1993 Aug;11(8):905-10. doi: 10.1038/nbt0893-905.
The Pichia pastoris heterologous gene expression system has been utilized to produce attractive levels of a variety of intracellular and extracellular proteins of interest. Recent advances in our understanding and application of the system have improved its utility even further. These advances include: (1) methods for the construction of P. pastoris strains with multiple copies of AOX1-promoter-driven expression cassettes; (2) mixed-feed culture strategies for high foreign protein volumetric productivity rates; (3) methods to reduce proteolysis of some products in high cell-density culture media; (4) tested procedures for purification of secreted products; and (5) detailed information on the structures of N-linked oligosaccharides on P. pastoris secreted proteins. In this review, these advances along with basic features of the P. pastoris system are described and discussed.
毕赤酵母异源基因表达系统已被用于生产大量具有吸引力的各种细胞内和细胞外目标蛋白。我们对该系统的理解和应用方面的最新进展进一步提高了其效用。这些进展包括：（1）构建具有多个AOX1启动子驱动表达盒拷贝的毕赤酵母菌株的方法；（2）用于提高外源蛋白体积生产率的混合进料培养策略；（3）在高细胞密度培养基中减少某些产物蛋白水解的方法；（4）分泌产物的测试纯化程序；以及（5）毕赤酵母分泌蛋白上N-连接寡糖结构的详细信息。在本综述中，将描述和讨论这些进展以及毕赤酵母系统的基本特征。
