Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
Appl Microbiol Biotechnol. 2010 Sep;88(1):75-86. doi: 10.1007/s00253-010-2718-9. Epub 2010 Jun 24.
Extracellular accumulation of recombinant proteins in the culture medium of Escherichia coli is desirable but difficult to obtain. The inner or cytoplasmic membrane and the outer membrane of E. coli are two barriers for releasing recombinant proteins expressed in the cytoplasm into the culture medium. Even if recombinant proteins have been exported into the periplasm, a space between the outer membrane and the inner membrane, the outer membrane remains the last barrier for their extracellular release. However, when E. coli was cultured in a particular defined medium, recombinant proteins exported into the periplasm could diffuse into the culture medium automatically. If a nonionic detergent, Triton X-100, was added in the medium, recombinant proteins expressed in the cytoplasm could also be released into the culture medium. It was then that extracellular accumulation of recombinant proteins could be obtained by exporting them into the periplasm or releasing them from the cytoplasm with Triton X-100 addition. The tactics described herein provided simple and valuable methods for achieving extracellular production of recombinant proteins in E. coli.
大肠杆菌培养物中重组蛋白的细胞外积累是可取的，但很难实现。大肠杆菌的内膜和外膜是将细胞质中表达的重组蛋白释放到培养基中的两个障碍。即使重组蛋白已被输出到周质（外膜和内膜之间的空间）中，外膜仍然是其细胞外释放的最后一道屏障。然而，当大肠杆菌在特定的限定培养基中培养时，输出到周质的重组蛋白可以自动扩散到培养基中。如果在培养基中加入非离子型去污剂 Triton X-100，也可以将细胞质中表达的重组蛋白释放到培养基中。然后，可以通过将它们输出到周质中或用 Triton X-100 从细胞质中释放来实现重组蛋白的细胞外积累。本文所述的策略为在大肠杆菌中实现重组蛋白的细胞外生产提供了简单而有价值的方法。
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