Secretion of 2,3-dihydroxyisovalerate as a limiting factor for isobutanol production in Saccharomyces cerevisiae

FEMS Yeast Research
Wesley Cardoso GenerosoEckhard Boles

Abstract

Isobutanol is a superior biofuel compared to ethanol, and it is naturally produced by yeasts. Previously, Saccharomyces cerevisiae has been genetically engineered to improve isobutanol production. We found that yeast cells engineered for a cytosolic isobutanol biosynthesis secrete large amounts of the intermediate 2,3-dihydroxyisovalerate (DIV). This indicates that the enzyme dihydroxyacid dehydratase (Ilv3) is limiting the isobutanol pathway and/or yeast exhibit effective transport systems for the secretion of the intermediate, competing with isobutanol synthesis. Moreover, we found that DIV cannot be taken up by the cells again. To identify the responsible transporters, microarray analysis was performed with a DIV producing strain compared to a wild type. Altogether, 19 genes encoding putative transporters were upregulated under DIV-producing conditions. Thirteen of these were deleted together with five homologous genes. A yro2 mrh1 deletion strain showed reduced DIV secretion, while a hxt5 deletion mutant showed increased isobutanol production. However, a strain deleted for all the 18 genes secreted even slightly increased amounts of the intermediates and less isobutanol. The lactate transporter Jen1 turned out to transport ...Continue Reading

References

Jan 15, 1997·Nucleic Acids Research·K R OldenburgC Paddon
Aug 1, 1997·FEMS Microbiology Reviews·E Boles, C P Hollenberg
Apr 10, 1999·Journal of Bacteriology·M CasalC Leão
Dec 5, 2000·Molecular Biology of the Cell·A P GaschP O Brown
Sep 5, 2002·Microbiology and Molecular Biology Reviews : MMBR·Lee R LyndIsak S Pretorius
Sep 5, 2002·Yeast·René VerwaalJohannes Boonstra
Jun 28, 2005·Nucleic Acids Research·Andreas GroteDieter Jahn
Oct 10, 2007·Biotechnology Journal·Peter Dürre
Feb 12, 2008·Applied and Environmental Microbiology·Beate Wiedemann, Eckhard Boles
Sep 2, 2008·FEMS Microbiology Reviews·Margarida CasalIsabel Soares-Silva
Apr 14, 2009·Nature Methods·Daniel G GibsonHamilton O Smith
Jun 11, 2010·Applied Microbiology and Biotechnology·Christian WeberEckhard Boles
Sep 24, 2010·Molecular Biology of the Cell·Joseph R McDermottZijuan Liu
Dec 7, 2010·Applied and Environmental Microbiology·Suvarna Dasari, Ralf Kölling
Nov 29, 2012·Bioresource Technology·Ethan I Lan, James C Liao
May 1, 2013·Current Opinion in Chemical Biology·Nicolaas A BuijsJens Nielsen
Apr 8, 2014·Proceedings of the National Academy of Sciences of the United States of America·Alexander FarwickEckhard Boles
Oct 7, 2014·Current Opinion in Biotechnology·Wesley Cardoso GenerosoEckhard Boles
Dec 17, 2014·Applied Microbiology and Biotechnology·Akiko TakabatakeShingo Izawa
Jun 26, 2015·Yeast·Uroš Petrovič
Mar 2, 2016·International Journal of Molecular Sciences·Danuza Nogueira MoysésFernando Araripe Gonçalves Torres
Jun 22, 2016·Journal of Microbiological Methods·Wesley Cardoso GenerosoEckhard Boles

❮ Previous
Next ❯

Citations

Nov 22, 2018·Frontiers in Microbiology·Akaraphol WatcharawipasHiroshi Takagi
Jun 11, 2020·Applied Microbiology and Biotechnology·Yike WangJian Hao
Dec 15, 2020·FEMS Microbiology Reviews·Justyna Ruchala, Andriy A Sibirny

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biofuels (ASM)

Biofuels are produced through contemporary processes from biomass rather than geological processes involved in fossil fuel formation. Examples include biodiesel, green diesel, biogas, etc. Discover the latest research on biofuels in this feed.