Cos Oriol, Serrano Alicia, Montesinos José Luis, Ferrer Pau, Cregg James M, Valero Francisco
Departament d'Enginyeria Química, ETSE, Universitat Autònoma de Barcelona, 08193 Bellaterra (Barcelona), Spain.
J Biotechnol. 2005 Apr 6;116(4):321-35. doi: 10.1016/j.jbiotec.2004.12.010.
An important number of heterologous proteins have been produced in the methylotrophic yeast Pichia pastoris using the alcohol oxidase promoter. Two factors that drastically influence protein production and cultivation process development in this system are gene dosage and methanol assimilation capacity of the host strain (Mut phenotype). Using a battery of four strains which secrete a Rhizopus oryzae lipase (ROL), the combined effects of gene dosage and Mut phenotype on recombinant protein production in Pichia pastoris was studied in fed-batch cultures. Regarding the effect of phenotype, the specific productivity and the Y(P/X) were 1.29- and 2.34-fold higher for Mut(s)ROL single copy strain than for Mut+ROL single copy strain. On the contrary, the productivity of Mut+ROL single copy strain was 1.34-fold higher than Mut(s)ROL single copy strain. An increase in ROL gene dosage seems to negatively affect cell's performance in bioreactor cultures, particularly in Mut(s) strains. Overall, the Mut(s) strain may be still advantageous to use because it allows for easier process control strategies.
利用醇氧化酶启动子，已经在甲基营养型酵母毕赤酵母中生产了大量的异源蛋白。在该系统中，对蛋白生产和培养过程发展有重大影响的两个因素是基因剂量和宿主菌株的甲醇同化能力（Mut表型）。使用一组分泌米根霉脂肪酶（ROL）的四种菌株，在补料分批培养中研究了基因剂量和Mut表型对毕赤酵母中重组蛋白生产的综合影响。关于表型的影响，Mut(s)ROL单拷贝菌株的比生产率和Y(P/X)分别比Mut+ROL单拷贝菌株高1.29倍和2.34倍。相反，Mut+ROL单拷贝菌株的生产率比Mut(s)ROL单拷贝菌株高1.34倍。ROL基因剂量的增加似乎对生物反应器培养中的细胞性能有负面影响，特别是在Mut(s)菌株中。总体而言，Mut(s)菌株可能仍然有利于使用，因为它允许采用更简便的过程控制策略。
