Mutagenesis of the Bacterial RNA Polymerase Alpha Subunit for Improvement of Complex Phenotypes
Mutagenesis of the Bacterial RNA Polymerase Alpha Subunit for Improvement of Complex Phenotypes
复制标题
用于改善复杂表型的细菌 RNA 聚合酶 α 亚基的诱变
DOI：
10.1128/aem.01888-08
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发表时间：
2009-05-01
影响因子：
4.4
通讯作者：
Stephanopoulos, Gregory
中科院分区：
文献类型：
--
作者：
Klein-Marcuschamer, Daniel;Santos, Christine Nicole S.;Stephanopoulos, Gregory
菌株工程的组合或随机方法已广泛用于改善多基因表型和其他性状，但其潜在机制尚未完全了解。尽管传统上首选的方法是诱变和选择，但我们的实验室已成功使用突变转录因子（在转录过程中指导 RNA 聚合酶 (RNAP)）来设计微生物细胞中的复杂表型。在这里，我们表明还可以通过改变 RNAP 核心酶本身，特别是通过细菌聚合酶 α 亚基的诱变来赋予新的表型。我们展示了使用该工具来提高大肠杆菌对丁醇和其他溶剂的耐受性，并提高两种商业相关产品（L-酪氨酸和透明质酸）的滴度。此外，我们还探索了产生耐溶剂突变体的潜在生理变化。
Combinatorial or random methods for strain engineering have been extensively used for the improvement of multigenic phenotypes and other traits for which the underlying mechanism is not fully understood. Although the preferred method has traditionally been mutagenesis and selection, our laboratory has successfully used mutant transcription factors, which direct the RNA polymerase (RNAP) during transcription, to engineer complex phenotypes in microbial cells. Here, we show that it is also possible to impart new phenotypes by altering the RNAP core enzyme itself, in particular through mutagenesis of the alpha subunit of the bacterial polymerase. We present the use of this tool for improving tolerance of Escherichia coli to butanol and other solvents and for increasing the titers of two commercially relevant products, L-tyrosine and hyaluronic acid. In addition, we explore the underlying physiological changes that give rise to the solvent-tolerant mutant.