Murby M, Uhlén M, Ståhl S
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Protein Expr Purif. 1996 Mar;7(2):129-36. doi: 10.1006/prep.1996.0018.
Proteolytic degradation of recombinant proteins represents a major problem related to production of gene products in heterologous hosts. Recombinant DNA technology offers several alternative strategies for stabilization of expressed gene products. These strategies can often give dramatic stabilization effects and can be combined with strategies involving optimization of fermentation conditions or downstream processing schemes. In this review, various genetic approaches to improve the stability of recombinant proteins will be discussed, including (i) choice of host cell strain, (ii) product localization, (iii) use of gene fusion partners, and (iv) product engineering. In addition, the solubility of the gene product can be influenced by factors such as growth temperature, promoter strength, fusion partners, and site-directed changes. Altogether, a battery of approaches can be used to obtain stable gene products.
重组蛋白的蛋白水解降解是异源宿主中基因产物生产相关的一个主要问题。重组DNA技术为稳定表达的基因产物提供了几种替代策略。这些策略通常能产生显著的稳定效果，并且可以与涉及优化发酵条件或下游加工方案的策略相结合。在本综述中，将讨论各种提高重组蛋白稳定性的遗传方法，包括（i）宿主细胞菌株的选择，（ii）产物定位，（iii）基因融合伙伴的使用，以及（iv）产物工程。此外，基因产物的溶解度会受到生长温度、启动子强度、融合伙伴和定点改变等因素的影响。总之，可以使用一系列方法来获得稳定的基因产物。
