Chemical genomic guided engineering of gamma-valerolactone tolerant yeast

Autor: Joshua J. Coon, Dan Xie, Mick McGee, Chad L. Myers, Yaoping Zhang, Jose Serate, Jeff S. Piotrowski, Quinn Dickinson, Alan Higbee, Li Hinchman, Robert Landick, Alexander S. Hebert, Scott Bottoms
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Proteomics
0301 basic medicine
Carboxy-Lyases
lcsh:QR1-502
Xylose
Lignin
Applied Microbiology and Biotechnology
7. Clean energy
lcsh:Microbiology
Lactones
chemistry.chemical_compound
immune system diseases
Ergosterol
Biomass
2. Zero hunger
chemistry.chemical_classification
0303 health sciences
biology
Strain (chemistry)
Genomics
Biocatalysts
Lignocellulosic
surgical procedures
operative
Biochemistry
Genetic Engineering
Biotechnology
Saccharomyces cerevisiae
Bioengineering
Chemical genomics
Hydrolysate
03 medical and health sciences
Biofuel
Drug Resistance
Fungal
Gamma-valerolactone
030304 developmental biology
Ethanol
030306 microbiology
Research
biology.organism_classification
Yeast
gamma-Valerolactone
030104 developmental biology
Enzyme
chemistry
Biofuels
Fermentation
Mutation
Biocatalysis
Zdroj: Microbial Cell Factories, Vol 17, Iss 1, Pp 1-12 (2018)
Microbial Cell Factories
ISSN: 1475-2859
DOI: 10.1186/s12934-017-0848-9
Popis: Background Gamma valerolactone (GVL) treatment of lignocellulosic bomass is a promising technology for degradation of biomass for biofuel production; however, GVL is toxic to fermentative microbes. Using a combination of chemical genomics with the yeast (Saccharomyces cerevisiae) deletion collection to identify sensitive and resistant mutants, and chemical proteomics to monitor protein abundance in the presence of GVL, we sought to understand the mechanism toxicity and resistance to GVL with the goal of engineering a GVL-tolerant, xylose-fermenting yeast. Results Chemical genomic profiling of GVL predicted that this chemical affects membranes and membrane-bound processes. We show that GVL causes rapid, dose-dependent cell permeability, and is synergistic with ethanol. Chemical genomic profiling of GVL revealed that deletion of the functionally related enzymes Pad1p and Fdc1p, which act together to decarboxylate cinnamic acid and its derivatives to vinyl forms, increases yeast tolerance to GVL. Further, overexpression of Pad1p sensitizes cells to GVL toxicity. To improve GVL tolerance, we deleted PAD1 and FDC1 in a xylose-fermenting yeast strain. The modified strain exhibited increased anaerobic growth, sugar utilization, and ethanol production in synthetic hydrolysate with 1.5% GVL, and under other conditions. Chemical proteomic profiling of the engineered strain revealed that enzymes involved in ergosterol biosynthesis were more abundant in the presence of GVL compared to the background strain. The engineered GVL strain contained greater amounts of ergosterol than the background strain. Conclusions We found that GVL exerts toxicity to yeast by compromising cellular membranes, and that this toxicity is synergistic with ethanol. Deletion of PAD1 and FDC1 conferred GVL resistance to a xylose-fermenting yeast strain by increasing ergosterol accumulation in aerobically grown cells. The GVL-tolerant strain fermented sugars in the presence of GVL levels that were inhibitory to the unmodified strain. This strain represents a xylose fermenting yeast specifically tailored to GVL produced hydrolysates. Electronic supplementary material The online version of this article (10.1186/s12934-017-0848-9) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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