Recombinant protein production associated growth inhibition results mainly from transcription and not from translation
Recombinant protein production associated growth inhibition results mainly from transcription and not from translation
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DOI：
10.1186/s12934-020-01343-y
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发表时间：
2020-04-06
影响因子：
6.4
通讯作者：
Rinas, Ursula
中科院分区：
文献类型：
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作者：
Li, Zhaopeng;Rinas, Ursula
背景重组蛋白的生产可能对宿主生物体造成压力。应激的程度由重组转录物和蛋白质的特定性质、转录和翻译速率以及生产过程中遇到的环境条件决定。结果T7启动子调控的人碱性成纤维细胞生长因子2（hFGF-2）和绿色荧光蛋白（GFP）基因的转录和翻译对生产宿主大肠杆菌生长特性的影响。coli BL 21（DE 3）中的表达。这是通过使用其中启动子区或核糖体结合位点或两者都被去除的表达载体来完成的。它表明，已经转录没有蛋白质翻译对宿主细胞施加代谢负担。在最好的情况下，将转录物翻译成大量正确折叠的蛋白质对细胞生长没有任何影响，例如在Luria-Bertani培养基中高水平产生GFP。然而，如果翻译与蛋白质折叠相关的问题（例如包涵体形成）有关，则翻译似乎有助于代谢负担。结论重组蛋白生产的代谢负荷主要来自转录，但可通过翻译和翻译后的过程（如蛋白质折叠和降解、热休克反应）得到增强。
Background Recombinant protein production can be stressful to the host organism. The extent of stress is determined by the specific properties of the recombinant transcript and protein, by the rates of transcription and translation, and by the environmental conditions encountered during the production process. Results The impact of the transcription of the T7-promoter controlled genes encoding human basic fibroblast growth factor (hFGF-2) and green fluorescent protein (GFP) as well as the translation into the recombinant protein on the growth properties of the production host E. coli BL21(DE3) were investigated. This was done by using expression vectors where the promoter region or the ribosome binding site(s) or both were removed. It is shown that already transcription without protein translation imposes a metabolic burden on the host cell. Translation of the transcript into large amounts of a properly folded protein does not show any effect on cell growth in the best case, e.g. high-level production of GFP in Luria-Bertani medium. However, translation appears to contribute to the metabolic burden if it is connected to protein folding associated problems, e.g. inclusion body formation. Conclusion The so-called metabolic burden of recombinant protein production is mainly attributed to transcription but can be enhanced through translation and those processes following translation (e.g. protein folding and degradation, heat-shock responses).