Ethanol tolerance in engineered strains of Clostridium thermocellum.
Acetaldehyde dehydrogenase
Acetivibrio thermocellus
AdhE
Alcohol dehydrogenase
Biofuel
Hungateiclostridium thermocellum
Next-generation sequencing
Ruminiclostridium thermocellum
Tolerance
Whole genome sequencing
Journal
Biotechnology for biofuels and bioproducts
ISSN: 2731-3654
Titre abrégé: Biotechnol Biofuels Bioprod
Pays: England
ID NLM: 9918300888906676
Informations de publication
Date de publication:
14 Sep 2023
14 Sep 2023
Historique:
received:
07
07
2023
accepted:
05
08
2023
medline:
15
9
2023
pubmed:
15
9
2023
entrez:
14
9
2023
Statut:
epublish
Résumé
Clostridium thermocellum is a natively cellulolytic bacterium that is promising candidate for cellulosic biofuel production, and can produce ethanol at high yields (75-80% of theoretical) but the ethanol titers produced thus far are too low for commercial application. In several strains of C. thermocellum engineered for increased ethanol yield, ethanol titer seems to be limited by ethanol tolerance. Previous work to improve ethanol tolerance has focused on the WT organism. In this work, we focused on understanding ethanol tolerance in several engineered strains of C. thermocellum. We observed a tradeoff between ethanol tolerance and production. Adaptation for increased ethanol tolerance decreases ethanol production. Second, we observed a consistent genetic response to ethanol stress involving mutations at the AdhE locus. These mutations typically reduced NADH-linked ADH activity. About half of the ethanol tolerance phenotype could be attributed to the elimination of NADH-linked activity based on a targeted deletion of adhE. Finally, we observed that rich growth medium increases ethanol tolerance, but this effect is eliminated in an adhE deletion strain. Together, these suggest that ethanol inhibits growth and metabolism via a redox-imbalance mechanism. The improved understanding of mechanisms of ethanol tolerance described here lays a foundation for developing strains of C. thermocellum with improved ethanol production.
Identifiants
pubmed: 37710260
doi: 10.1186/s13068-023-02379-z
pii: 10.1186/s13068-023-02379-z
pmc: PMC10503014
doi:
Types de publication
Journal Article
Langues
eng
Pagination
137Subventions
Organisme : NCI NIH HHS
ID : P30 CA023108
Pays : United States
Informations de copyright
© 2023. BioMed Central Ltd., part of Springer Nature.
Références
Metab Eng. 2013 May;17:1-11
pubmed: 23376594
Biotechnol Biofuels. 2017 Nov 30;10:276
pubmed: 29213320
Microbiol Mol Biol Rev. 2002 Sep;66(3):506-77, table of contents
pubmed: 12209002
Science. 2006 Dec 8;314(5805):1565-8
pubmed: 17158319
Genetics. 2007 Mar;175(3):1479-87
pubmed: 17194785
PLoS One. 2014 Feb 07;9(2):e86389
pubmed: 24516531
BMC Genomics. 2012 Jul 23;13:336
pubmed: 22823947
J Bacteriol. 2012 Mar;194(5):1145-57
pubmed: 22210771
Sci Rep. 2019 Feb 11;9(1):1736
pubmed: 30741948
J Bacteriol. 1988 Feb;170(2):775-80
pubmed: 2828323
Curr Genet. 2016 Aug;62(3):475-80
pubmed: 26758993
Appl Microbiol Biotechnol. 2010 Jul;87(3):829-45
pubmed: 20464391
Biotechnol Biofuels. 2016 Jun 16;9:125
pubmed: 27313661
Metab Eng. 2013 Jan;15:124-33
pubmed: 23164575
Appl Microbiol Biotechnol. 2007 Feb;74(2):422-32
pubmed: 17124583
Commun Biol. 2020 Jun 10;3(1):298
pubmed: 32523125
J Bacteriol. 1980 Nov;144(2):569-78
pubmed: 7430065
J Bacteriol. 1988 Jun;170(6):2809-15
pubmed: 3372483
Proc Natl Acad Sci U S A. 2014 Jun 24;111(25):E2576-85
pubmed: 24927582
J Bacteriol. 2015 Apr;197(8):1386-93
pubmed: 25666131
Microb Cell Fact. 2022 Dec 25;21(1):273
pubmed: 36567317
Biotechnol Biofuels. 2015 Feb 12;8:20
pubmed: 25763101
FEBS Lett. 1983 Jun 13;156(2):303-6
pubmed: 6343121
Methods Enzymol. 2012;510:317-30
pubmed: 22608734
Appl Microbiol Biotechnol. 2009 Apr;82(5):929-39
pubmed: 19221734
Appl Environ Microbiol. 2011 Dec;77(23):8288-94
pubmed: 21965408
Appl Environ Microbiol. 1980 Sep;40(3):571-7
pubmed: 7425617
Proc Natl Acad Sci U S A. 2011 Aug 16;108(33):13752-7
pubmed: 21825121
Biotechnol Prog. 2003 Mar-Apr;19(2):612-23
pubmed: 12675606
Biotechnol Biofuels. 2018 Sep 06;11:242
pubmed: 30202437
J Biol Chem. 1978 Jul 25;253(14):5048-55
pubmed: 353047
J Bacteriol. 1984 Jan;157(1):233-9
pubmed: 6360997
Mol Syst Biol. 2010 Jun 8;6:378
pubmed: 20531407
Biochim Biophys Acta. 1982 Dec 8;693(1):195-204
pubmed: 7150588
Crit Rev Biotechnol. 1990;9(4):305-19
pubmed: 2178781
J Biol Chem. 1985 Jun 25;260(12):7442-51
pubmed: 3997879
Metab Eng Commun. 2019 Nov 28;10:e00116
pubmed: 31890588
Biotechnol Biofuels. 2020 Mar 10;13:40
pubmed: 32175007
Front Microbiol. 2015 Feb 18;6:103
pubmed: 25741329
PLoS Genet. 2015 Nov 06;11(11):e1005635
pubmed: 26545090
Mol Biol Evol. 2017 Nov 1;34(11):2927-2943
pubmed: 28961727
Crit Rev Microbiol. 1986;13(3):219-80
pubmed: 3533426
Appl Microbiol Biotechnol. 2011 Nov;92(3):641-52
pubmed: 21874277
BMC Evol Biol. 2015 Sep 03;15:180
pubmed: 26334309
J Bacteriol. 2015 Aug 1;197(15):2610-9
pubmed: 26013492
Biotechnol Biofuels. 2016 Jun 02;9:116
pubmed: 27257435
Metab Eng. 2019 Sep;55:161-169
pubmed: 31220663
J Mol Biol. 1990 Oct 5;215(3):403-10
pubmed: 2231712