Extracellular accumulation of recombinant protein by Escherichia coli in a defined medium
Extracellular accumulation of recombinant protein by Escherichia coli in a defined medium
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DOI：
10.1007/s00253-010-2718-9
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发表时间：
2010-09-01
影响因子：
5
通讯作者：
Fu, Xiang-Yang
中科院分区：
文献类型：
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作者：
Fu, Xiang-Yang
重组蛋白在大肠杆菌培养基中的细胞外积累是期望的，但难以获得。E.大肠杆菌是将在细胞质中表达的重组蛋白释放到培养基中的两个屏障。即使重组蛋白已经输出到周质中，即外膜和内膜之间的空间，外膜仍然是它们在细胞外释放的最后屏障。然而，当E.大肠杆菌在特定的培养基中培养，输出到周质中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去污剂Triton X-100，在细胞质中表达的重组蛋白也可以释放到培养基中。然后，可以通过将重组蛋白输出到周质中或通过添加Triton X-100将其从细胞质中释放来获得重组蛋白的胞外积累。本文所述的策略为在大肠杆菌中实现重组蛋白的胞外生产提供了简单而有价值的方法。杆菌
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.