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高通量筛选辛酸产生菌文库可用于检测提高. 产量的新靶点

High-Throughput Screening of an Octanoic Acid Producer Strain Library Enables Detection of New Targets for Increasing Titers in .

机构信息

Institute of Molecular Biosciences, Faculty of Biological Sciences, Goethe University Frankfurt, Max-von-Laue Straße 9, 60438 Frankfurt am Main, Germany.

Department of Biology, Computer-aided Synthetic Biology, Technical University Darmstadt, Schnittspahnstr. 1, 64287 Darmstadt, Germany.

出版信息

ACS Synth Biol. 2021 May 21;10(5):1077-1086. doi: 10.1021/acssynbio.0c00600. Epub 2021 May 12.

Abstract

Octanoic acid is an industrially relevant compound with applications in antimicrobials or as a precursor for biofuels. Microbial biosynthesis through yeast is a promising alternative to current unsustainable production methods. To increase octanoic acid titers in , we use a previously developed biosensor that is based on the octanoic acid responsive promotor coupled to GFP. We establish a biosensor strain amenable for high-throughput screening of an octanoic acid producer strain library. Through development, optimization, and execution of a high-throughput screening approach, we were able to detect two new genetic targets, and , which increased octanoic acid titers through combined overexpression by about 55% compared to the parental strain. Neither target has yet been reported to be involved in fatty acid biosynthesis. The presented methodology can be employed to screen any genetic library and thereby more genes involved in improving octanoic acid production can be detected in the future.

摘要

辛酸是一种具有工业应用价值的化合物,可用于抗菌剂或生物燃料的前体。通过酵母进行微生物生物合成是一种有前途的替代当前不可持续生产方法的方法。为了提高 的辛酸产量,我们使用了先前开发的生物传感器,该传感器基于与 GFP 偶联的辛酸响应启动子。我们建立了一种生物传感器菌株,可用于高通量筛选辛酸生产菌株文库。通过高通量筛选方法的开发、优化和执行,我们能够检测到两个新的遗传靶标 和 ,与亲本菌株相比,通过组合过表达将辛酸产量提高了约 55%。这两个靶标都尚未报道与脂肪酸生物合成有关。所提出的方法可用于筛选任何基因文库,从而可以在未来检测到更多参与提高辛酸产量的基因。

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