Amino acid stabilization for cell-free protein synthesis by modification of the Escherichia coli genome
Amino acid stabilization for cell-free protein synthesis by modification of the Escherichia coli genome
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DOI：
10.1016/j.ymben.2004.01.003
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发表时间：
2004-07-01
影响因子：
8.4
通讯作者：
Swartz, J
中科院分区：
文献类型：
--
作者：
Michel-Reydellet, N;Calhoun, K;Swartz, J
无细胞生物学提供了通过逆代谢工程评估和操纵微生物系统的独特机会。我们已经将这种方法应用于氨基酸代谢，这是无细胞生物学中限制蛋白质合成反应的系统之一。四种氨基酸（精氨酸、色氨酸、丝氨酸和半胱氨酸）在各种条件下的3小时分批无细胞蛋白质合成反应期间耗尽。通过修改用于制备细胞提取物的大肠杆菌菌株的基因组，我们看到精氨酸、色氨酸和丝氨酸的显着稳定。然而，半胱氨酸继续被降解。使用修饰的细胞提取物进行无细胞蛋白质合成，可以提高无半胱氨酸蛋白质外膜蛋白T（OmpT）的产量。(C)2004年爱思唯尔公司All rights reserved.
Cell-free biology provides a unique opportunity to assess and to manipulate microbial systems by inverse metabolic engineering. We have applied this approach to amino acid metabolism, one of the systems in cell-free biology that limits protein synthesis reactions. Four amino acids (arginine, tryptophan, serine and cysteine) are depleted during a 3-h batch cell-free protein synthesis reaction under various conditions. By modifying the genome of the Escherichia coli strain used to make the cell extract, we see significant stabilization of arginine, tryptophan and serine. Cysteine, however, continues to be degraded. Cell-free protein synthesis with the modified cell extract produces increased yields of the cysteine-free protein Outer Membrane Protein T (OmpT). (C) 2004 Elsevier Inc. All rights reserved.