The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum

Zheng, Tianyong; Lanahan, Anthony A.; Lynd, Lee R.; Olson, Daniel G.; Blanchard, ed., Jeffrey L.

doi:10.1371/journal.pone.0195143

Title: The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum

Journal Article · Thu Apr 05 00:00:00 EDT 2018 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0195143· OSTI ID:1432066

; Lanahan, Anthony A.; Lynd, Lee R.;

; Blanchard, ed., Jeffrey L.

Thermoanaerobacterium saccharolyticum is a thermophilic anaerobe that has been engineered to produce high amounts of ethanol, reaching ~90% theoretical yield at a titer of 70 g/L. Here we report the physiological changes that occur upon deleting the redox-sensing transcriptional regulator Rex in wild type T. saccharolyticum: a single deletion of rex resulted in a two-fold increase in ethanol yield (from 40% to 91% theoretical yield), but the resulting strains grew only about a third as fast as the wild type strain. Deletion of the rex gene also had the effect of increasing expression of alcohol dehydrogenase genes, adhE and adhA. After several serial transfers, the ethanol yield decreased from an average of 91% to 55%, and the growth rates had increased. We performed whole-genome resequencing to identify secondary mutations in the Δrex strains adapted for faster growth. In several cases, secondary mutations had appeared in the adhE gene. Furthermore, in these strains the NADH-linked alcohol dehydrogenase activity was greatly reduced. Complementation studies were done to reintroduce rex into the Δ strains: reintroducing rex decreased ethanol yield to below wild type levels in the Δrex strain without adhE mutations, but did not change the ethanol yield in the Δrex strain where an adhE mutation occurred.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1432066

Alternate ID(s):: OSTI ID: 1627859

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Vol. 13 Journal Issue: 4; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 8 works

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