Jahic Mehmedalija, Veide Andres, Charoenrat Theppanya, Teeri Tuula, Enfors Sven-Olof
Department of Bioprocess Technology, School of Biotechnology, Royal Institute of Technology (KTH), S 10691 Stockholm, Sweden.
Biotechnol Prog. 2006 Nov-Dec;22(6):1465-73. doi: 10.1021/bp060171t.
Developments in process techniques for production and recovery of heterologous proteins with Pichia pastoris are presented. Limitations for the standard techniques are described, and alternative techniques that solve the limitations problems are reviewed together with the methods that resulted in higher productivity of the P. pastoris processes. The main limitations are proteolysis of the secreted products and cell death in the high cell density bioreactor cultures. As a consequence, both low productivity and lower quality of the feedstock for downstream processing are achieved in processes hampered with these problems. Methods for exploring proteolysis and cell death are also presented. Solving the problems makes the conditions for downstream processing superior for the P. pastoris expression systems compared to other systems, which either need complex media or rely on intracellular production. These improved conditions allow for interfacing of cultivation with downstream processing in an integrated fashion.
本文介绍了利用毕赤酵母生产和回收异源蛋白的工艺技术进展。描述了标准技术的局限性，并综述了能够解决这些局限性问题的替代技术，以及提高毕赤酵母生产工艺生产率的方法。主要局限性在于分泌产物的蛋白水解作用以及高细胞密度生物反应器培养中的细胞死亡。因此，在受这些问题困扰的工艺中，生产率较低且下游加工的原料质量也较低。本文还介绍了探究蛋白水解作用和细胞死亡的方法。与其他需要复杂培养基或依赖细胞内生产的系统相比，解决这些问题使毕赤酵母表达系统的下游加工条件更为优越。这些改善的条件允许以整合的方式将培养与下游加工衔接起来。
