Modulating Membrane Composition Alters Free Fatty Acid Tolerance in Escherichia coli

Rebecca M. Lennen, Brian F. Pfleger

Research output: Contribution to journalJournal articleResearchpeer-review

347 Downloads (Pure)

Abstract

Microbial synthesis of free fatty acids (FFA) is a promising strategy for converting renewable sugars to advanced biofuels and oleochemicals. Unfortunately, FFA production negatively impacts membrane integrity and cell viability in Escherichia coli, the dominant host in which FFA production has been studied. These negative effects provide a selective pressure against FFA production that could lead to genetic instability at industrial scale. In prior work, an engineered E. coli strain harboring an expression plasmid for the Umbellularia californica acyl-acyl carrier protein (ACP) thioesterase was shown to have highly elevated levels of unsaturated fatty acids in the cell membrane. The change in membrane content was hypothesized to be one underlying cause of the negative physiological effects associated with FFA production. In this work, a connection between the regulator of unsaturated fatty acid biosynthesis in E. coli, FabR, thioesterase expression, and unsaturated membrane content was established. A strategy for restoring normal membrane saturation levels and increasing tolerance towards endogenous production of FFAs was implemented by modulating acyl-ACP pools with a second thioesterase (from Geobacillus sp. Y412MC10) that primarily targets medium chain length, unsaturated acyl-ACPs. The strategy succeeded in restoring membrane content and improving viability in FFA producing E. coli while maintaining FFA titers. However, the restored fitness did not increase FFA productivity, indicating the existence of additional metabolic or regulatory barriers.
Original languageEnglish
Article numbere54031
JournalP L o S One
Volume8
Issue number1
Number of pages10
ISSN1932-6203
Publication statusPublished - 2013
Externally publishedYes

Fingerprint Dive into the research topics of 'Modulating Membrane Composition Alters Free Fatty Acid Tolerance in Escherichia coli'. Together they form a unique fingerprint.

Cite this