Improving olefin tolerance and production in E. coli using native and evolved AcrB.
Improving olefin tolerance and production in E. coli using native and evolved AcrB.
复制标题
DOI：
10.1002/bit.25511
复制
发表时间：
2015-05
影响因子：
3.8
通讯作者：
Mukhopadhyay, Aindrila
中科院分区：
文献类型：
--
作者：
Mingardon, Florence;Clement, Camille;Hirano, Kathleen;Nhan, Melissa;Luning, Eric G.;Chanal, Angelique;Mukhopadhyay, Aindrila
微生物可以被设计用于生产聚合物工业中使用的化学品。然而，许多此类目标化合物会抑制微生物生长，并可能相应地限制生产水平。在这里，我们重点关注生物塑料前体的化合物，特别是苯乙烯和代表性的 α-烯烃； 1-己烯、1-辛烯和1-壬烯。我们评估了大肠杆菌外排泵 AcrAB-TolC 在增强对这些烯烃化合物的耐受性方面的作用。 AcrAB-TolC 参与大肠杆菌对所有四种化合物的耐受性。苯乙烯和1-己烯对大肠杆菌都有剧毒。苯乙烯是一种模型塑料前体，在大肠杆菌中已建立生产路线（McKenna 和 Nielsen，2011）。尽管我们的数据表明 AcrAB-TolC 对于其最佳生产很重要，但我们观察到大肠杆菌中苯乙烯生产存在强烈的负选择。因此，我们使用1-己烯作为模型化合物来实施定向进化策略，以进一步提高对该α-烯烃的耐受表型。我们重点关注 AcrB（已知负责底物结合的内膜结构域）的优化，并发现了几个导致耐受性改善的突变（A279T、Q584R、F617L、L822P、F927S 和 F1033Y）。其中一些突变也可以以协同方式组合。我们的研究表明，外排泵是烯烃宿主工程中的一种重要机制，并且可以使用定向进化等策略进一步改进，以提高耐受性和潜在产量。生物技术。生物工程。 2015；112：879–888。 © 2015 作者。生物技术和生物工程由 John Wiley & periodicals, Inc. 出版
Microorganisms can be engineered for the production of chemicals utilized in the polymer industry. However many such target compounds inhibit microbial growth and might correspondingly limit production levels. Here, we focus on compounds that are precursors to bioplastics, specifically styrene and representative alpha-olefins; 1-hexene, 1-octene, and 1-nonene. We evaluated the role of the Escherichia coli efflux pump, AcrAB-TolC, in enhancing tolerance towards these olefin compounds. AcrAB-TolC is involved in the tolerance towards all four compounds in E. coli. Both styrene and 1-hexene are highly toxic to E. coli. Styrene is a model plastics precursor with an established route for production in E. coli (McKenna and Nielsen, 2011). Though our data indicates that AcrAB-TolC is important for its optimal production, we observed a strong negative selection against the production of styrene in E. coli. Thus we used 1-hexene as a model compound to implement a directed evolution strategy to further improve the tolerance phenotype towards this alpha-olefin. We focused on optimization of AcrB, the inner membrane domain known to be responsible for substrate binding, and found several mutations (A279T, Q584R, F617L, L822P, F927S, and F1033Y) that resulted in improved tolerance. Several of these mutations could also be combined in a synergistic manner. Our study shows efflux pumps to be an important mechanism in host engineering for olefins, and one that can be further improved using strategies such as directed evolution, to increase tolerance and potentially production. Biotechnol. Bioeng. 2015;112: 879–888. © 2015 The Authors. Biotechnology and Bioengineering Published by John Wiley & Periodicals, Inc.
登录
查看更多内容
影响因子：
3.6
作者：
Husain F;Nikaido H
通讯作者：
Nikaido H
影响因子：
3.2
作者：
Bohnert, Juergen A.;Schuster, Sabine;Kern, Winfried V.
通讯作者：
Kern, Winfried V.
影响因子：
--
作者：
Jarboe LR;Zhang X;Wang X;Moore JC;Shanmugam KT;Ingram LO
通讯作者：
Ingram LO
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
DOI：
10.1073/pnas.1301358110
发表时间：
2013-05-07
影响因子：
11.1
作者：
Doshi, Rupak;Tuan Nguyen;Chang, Geoffrey
通讯作者：
Chang, Geoffrey