Use of E. coli for the production of a single protein.
Use of E. coli for the production of a single protein.
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DOI：
10.1007/978-1-61779-921-1_11
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Inouye, Masayori
中科院分区：
文献类型：
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作者：
Mao, Lili;Inouye, Masayori
大肠杆菌已广泛用于重组蛋白生产。在这里，我们介绍了一种新的大肠杆菌表达方法，即单一蛋白质生产（SPP）系统，其中大肠杆菌被转化为仅生产目标蛋白质的生物反应器。在 SPP 系统中，所有大肠杆菌细胞 mRNA 通过 MazF（一种 ACA 特异性 mRNA 干扰酶）的诱导而被消除，从而导致细胞生长完全停滞。然而，目标蛋白的 mRNA 被设计为不含 ACA 序列，因此能够抵抗 MazF 裂解。因此，SPP 系统是独特的，非常适合表达有毒蛋白质和掺入有毒氨基酸类似物。我们还证明，SPP 系统是一种用于 NMR 结构研究的经济有效的蛋白质生产方法，因为细胞培养物可以高度浓缩而不影响蛋白质产量。
E. coli has been widely used for recombinant protein production. Here, we introduce a novel expression method in E. coli, the Single Protein Production (SPP) system, in which E. coli is converted into a bioreactor producing only the target protein. In the SPP system, all E. coli cellular mRNAs are eliminated by the induction of MazF, an ACA-specific mRNA interferase, which results in complete cell growth arrest. However, the mRNA for a target protein was engineered to be devoid of ACA sequences, thus resistant to MazF cleavage. Therefore, the SPP system is unique and ideal for expression of toxic proteins and incorporation of toxic amino acid analogues. We have also demonstrated that the SPP system is a cost-effective protein production method for NMR structural studies because cell culture can be highly condensed without affecting protein yields.