Escherichia coli "TatExpress" strains super-secrete human growth hormone into the bacterial periplasm by the Tat pathway.
Escherichia coli "TatExpress" strains super-secrete human growth hormone into the bacterial periplasm by the Tat pathway.
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DOI：
10.1002/bit.26434
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发表时间：
2017-12
影响因子：
3.8
通讯作者：
Robinson C
中科院分区：
文献类型：
--
作者：
Browning DF;Richards KL;Peswani AR;Roobol J;Busby SJW;Robinson C
许多高价值的蛋白质通过一般分泌（Sec）途径分泌到大肠杆菌周质中，但基于Sec的生产底盘不能处理许多潜在的靶蛋白。达特途径提供了一个有前途的替代方案，因为它运输完全折叠的蛋白质;然而，产量太低，无法商业化使用。为了促进达特输出，我们通过在染色体tatABCD操纵子的上游引入强诱导型细菌启动子ptac，以驱动其在E.工业上常用的大肠杆菌菌株（例如，W3110和BL 21）。这种修饰显著改善了人生长激素（hGH）向细菌周质中的达特依赖性分泌，达到仅在1小时诱导后分泌的hGH就成为主要周质蛋白的程度。即使在摇瓶培养中，TatExpress菌株也积累超过30 mg L−1周质重组hGH。第二种靶蛋白，scFv，也显示在TatExpress菌株中以高得多的速率输出。
Numerous high‐value proteins are secreted into the Escherichia coli periplasm by the General Secretory (Sec) pathway, but Sec‐based production chassis cannot handle many potential target proteins. The Tat pathway offers a promising alternative because it transports fully folded proteins; however, yields have been too low for commercial use. To facilitate Tat export, we have engineered the TatExpress series of super‐secreting strains by introducing the strong inducible bacterial promoter, ptac, upstream of the chromosomal tatABCD operon, to drive its expression in E. coli strains commonly used by industry (e.g., W3110 and BL21). This modification significantly improves the Tat‐dependent secretion of human growth hormone (hGH) into the bacterial periplasm, to the extent that secreted hGH is the dominant periplasmic protein after only 1 hr induction. TatExpress strains accumulate in excess of 30 mg L−1 periplasmic recombinant hGH, even in shake flask cultures. A second target protein, an scFv, is also shown to be exported at much higher rates in TatExpress strains.
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影响因子：
3.5
作者：
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通讯作者：
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通讯作者：
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