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Title: PKA and HOG signaling contribute separable roles to anaerobic xylose fermentation in yeast engineered for biofuel production

Journal Article · · PLoS ONE
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Lignocellulosic biomass offers a sustainable source for biofuel production that does not compete with food-based cropping systems. Importantly, two critical bottlenecks prevent economic adoption: many industrially relevant microorganisms cannot ferment pentose sugars prevalent in lignocellulosic medium, leaving a significant amount of carbon unutilized. Furthermore, chemical biomass pretreatment required to release fermentable sugars generates a variety of toxins, which inhibit microbial growth and metabolism, specifically limiting pentose utilization in engineered strains. Here we dissected genetic determinants of anaerobic xylose fermentation and stress tolerance in chemically pretreated corn stover biomass, called hydrolysate. We previously revealed that loss-of-function mutations in the stress-responsive MAP kinase HOG1 and negative regulator of the RAS/Protein Kinase A (PKA) pathway, IRA2, enhances anaerobic xylose fermentation. However, these mutations likely reduce cells’ ability to tolerate the toxins present in lignocellulosic hydrolysate, making the strain especially vulnerable to it. We tested the contributions of Hog1 and PKA signaling via IRA2 or PKA negative regulatory subunit BCY1 to metabolism, growth, and stress tolerance in corn stover hydrolysate and laboratory medium with mixed sugars. We found mutations causing upregulated PKA activity increase growth rate and glucose consumption in various media but do not have a specific impact on xylose fermentation. In contrast, mutation of HOG1 specifically increased xylose usage. We hypothesized improving stress tolerance would enhance the rate of xylose consumption in hydrolysate. Surprisingly, increasing stress tolerance did not augment xylose fermentation in lignocellulosic medium in this strain background, suggesting other mechanisms besides cellular stress limit this strain’s ability for anaerobic xylose fermentation in hydrolysate.

Research Organization:
Univ. of Wisconsin, Madison, WI (United States); USDOE Bioenergy Research Centers (BRC) (United States). Great Lakes Bioenergy Research Center (GLBRC)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF); Wisconsin Alumni Research Foundation
Grant/Contract Number:
SC0018409; DGE-1747503
OSTI ID:
1514759
Alternate ID(s):
OSTI ID: 1612978
Journal Information:
PLoS ONE, Journal Name: PLoS ONE Vol. 14 Journal Issue: 5; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 7 works
Citation information provided by
Web of Science

References (82)

Inhibition effects of furfural on alcohol dehydrogenase, aldehyde dehydrogenase and pyruvate dehydrogenase journal April 2002
Conversion of Xylose to Ethanol by Recombinant Saccharomyces cerevisiae: Importance of Xylulokinase (XKS1) and Oxygen Availability journal July 2001
High-level functional expression of a fungal xylose isomerase: the key to efficient ethanolic fermentation of xylose by ? journal October 2003
Optimization of an acetate reduction pathway for producing cellulosic ethanol by engineered yeast: Optimization of the Acetate Reduction Pathway in Yeast journal September 2016
Simultaneously improving xylose fermentation and tolerance to lignocellulosic inhibitors through evolutionary engineering of recombinant Saccharomyces cerevisiae harbouring xylose isomerase journal January 2014
Pulsed addition of HMF and furfural to batch-grown xylose-utilizing Saccharomyces cerevisiae results in different physiological responses in glucose and xylose consumption phase journal January 2013
Unraveling the genetic basis of xylose consumption in engineered Saccharomyces cerevisiae strains journal December 2016
Rational and Evolutionary Engineering Approaches Uncover a Small Set of Genetic Changes Efficient for Rapid Xylose Fermentation in Saccharomyces cerevisiae journal February 2013
Inhibition of ethanol-producing yeast and bacteria by degradation products produced during pre-treatment of biomass journal August 2004
Nutrient-induced activation of trehalase in nutrient-starved cells of the yeast Saccharomyces cerevisiae: cAMP is not involved as second messenger journal October 1992
Kelch-repeat proteins interacting with the G  protein Gpa2 bypass adenylate cyclase for direct regulation of protein kinase A in yeast journal August 2006
In Vivo Phosphorylation of Ser 21 and Ser 83 during Nutrient-induced Activation of the Yeast Protein Kinase A (PKA) Target Trehalase journal November 2012
Expression of Gre2p improves tolerance of engineered xylose-fermenting Saccharomyces cerevisiae to glycolaldehyde under xylose metabolism journal July 2018
NeuCode Labels for Relative Protein Quantification journal June 2014
Simultaneous co-fermentation of mixed sugars: a promising strategy for producing cellulosic ethanol journal May 2012
Nutrient availability and the RAS/cyclic AMP pathway both induce expression of ribosomal protein genes in Saccharomyces cerevisiae but by different mechanisms. journal June 1995
Identification of furfural as a key toxin in lignocellulosic hydrolysates and evolution of a tolerant yeast strain journal November 2008
A Saccharomyces cerevisiae UAS element controlled by protein kinase A activates transcription in response to a variety of stress conditions. journal May 1993
Delayed Turnover of Unphosphorylated Ssk1 during Carbon Stress Activates the Yeast Hog1 Map Kinase Pathway journal September 2015
Molecular response of Saccharomyces cerevisiae wine and laboratory strains to high sugar stress conditions journal January 2011
Activity of glycolytic enzymes of Saccharomyces cerevisiae in the presence of acetic acid journal December 1990
Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant journal January 1990
Protein kinase A mediates growth-regulated expression of yeast ribosomal protein genes by modulating RAP1 transcriptional activity. journal March 1994
Exploring the Impact of Osmoadaptation on Glycolysis Using Time-Varying Response-Coefficients
  • KÜHn, Clemens; Petelenz, Elzbieta; Nordlander, Bodil
  • Proceedings of the 8th Annual International Workshop on Bioinformatics and Systems Biology (IBSB 2008), Genome Informatics 2008 https://doi.org/10.1142/9781848163003_0007
conference January 2012
Nucleocytoplasmic Distribution of Budding Yeast Protein Kinase A Regulatory Subunit Bcy1 Requires Zds1 and Is Regulated by Yak1-Dependent Phosphorylation of Its Targeting Domain journal January 2001
Network inference reveals novel connections in pathways regulating growth and defense in the yeast salt response journal May 2018
Engineering tolerance to industrially relevant stress factors in yeast cell factories journal June 2017
Mechanisms of protein kinase Sch9 regulating Bcy1 in Saccharomyces cerevisiae journal April 2012
Bcy1, the regulatory subunit of cAMP-dependent protein kinase in yeast, is differentially modified in response to the physiological status of the cell. journal October 1991
Alcoholic fermentation of carbon sources in biomass hydrolysates by Saccharomyces cerevisiae: current status journal October 2006
Stress-related challenges in pentose fermentation to ethanol by the yeast Saccharomyces cerevisiae journal February 2011
Effects of acetic acid on the kinetics of xylose fermentation by an engineered, xylose-isomerase-based Saccharomyces cerevisiae strain journal May 2009
Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast journal October 2013
Combining inhibitor tolerance and D-xylose fermentation in industrial Saccharomyces cerevisiae for efficient lignocellulose-based bioethanol production journal January 2013
Activation State of the Ras2 Protein and Glucose-induced Signaling in Saccharomyces cerevisiae journal August 2004
Leveraging Genetic-Background Effects in Saccharomyces cerevisiae To Improve Lignocellulosic Hydrolysate Tolerance journal July 2016
Phenolic Amides Are Potent Inhibitors of De Novo Nucleotide Biosynthesis journal June 2015
Nutritional Control of Nucleocytoplasmic Localization of cAMP-dependent Protein Kinase Catalytic and Regulatory Subunits in Saccharomyces cerevisiae journal January 2000
Response to Hyperosmotic Stress journal October 2012
Differential Effects of Substrate on Type I and Type II PKA Holoenzyme Dissociation † journal May 2004
Open Mass Spectrometry Search Algorithm journal October 2004
Quantitative Analysis of Glycerol Accumulation, Glycolysis and Growth under Hyper Osmotic Stress journal June 2013
The Yeast Cyclin-Dependent Kinase Routes Carbon Fluxes to Fuel Cell Cycle Progression journal May 2016
Carbon fluxes of xylose-consuming Saccharomyces cerevisiae strains are affected differently by NADH and NADPH usage in HMF reduction journal June 2009
Activation of trehalase during growth induction by nitrogen sources in the yeastSaccharomyces cerevisiae depends on the free catalytic subunits of camp-dependent protein kinase, but not on functional ras proteins journal August 1994
High Osmolarity Glycerol (HOG) Pathway-induced Phosphorylation and Activation of 6-Phosphofructo-2-kinase Are Essential for Glycerol Accumulation and Yeast Cell Proliferation under Hyperosmotic Stress journal March 2004
Genetically Engineered SaccharomycesYeast Capable of Effective Cofermentation of Glucose and Xylose journal May 1998
Overexpression of the yeast transcription activator Msn2 confers furfural resistance and increases the initial fermentation rate in ethanol production journal April 2012
COMPASS: A suite of pre- and post-search proteomics software tools for OMSSA journal February 2011
Screening and construction of Saccharomyces cerevisiae strains with improved multi-tolerance and bioethanol fermentation performance journal February 2011
Combinatorial control by the protein kinases PKA, PHO85 and SNF1 of transcriptional induction of the Saccharomyces cerevisiae GSY2 gene at the diauxic shift journal June 2004
Overexpression of RCK1 improves acetic acid tolerance in Saccharomyces cerevisiae journal February 2019
Optimal Growth and Ethanol Production from Xylose by Recombinant Saccharomyces cerevisiae Require Moderate D-Xylulokinase Activity journal January 2003
Glycerol formation during wine fermentation is mainly linked to Gpd1p and is only partially controlled by the HOG pathway journal January 2003
Positioning of cell growth and division after osmotic stress requires a map kinase pathway journal April 1994
Reserve carbohydrates metabolism in the yeast Saccharomyces cerevisiae journal January 2001
Exploiting the yeast stress-activated signaling network to inform on stress biology and disease signaling journal May 2015
In Vivo and in Vitro Phosphorylation of Two Isoforms of Yeast Pyruvate Kinase by Protein Kinase A journal June 2002
Nutrient sensing and signaling in the yeast Saccharomyces cerevisiae journal March 2014
Metabolic engineering of a xylose-isomerase-expressing strain for rapid anaerobic xylose fermentation journal February 2005
Re-assessment of YAP1 and MCR1 contributions to inhibitor tolerance in robust engineered Saccharomyces cerevisiae fermenting undetoxified lignocellulosic hydrolysate journal July 2014
Xylulokinase Overexpression in Two Strains of Saccharomyces cerevisiae Also Expressing Xylose Reductase and Xylitol Dehydrogenase and Its Effect on Fermentation of Xylose and Lignocellulosic Hydrolysate journal September 2001
Metabolic Activation of the HOG MAP Kinase Pathway by Snf1/AMPK Regulates Lipid Signaling at the Golgi: Metabolic Regulation Golgi PI(4)P journal September 2012
Death by a thousand cuts: the challenges and diverse landscape of lignocellulosic hydrolysate inhibitors journal March 2014
Multifaceted characterization of cell wall decomposition products formed during ammonia fiber expansion (AFEX) and dilute acid based pretreatments journal November 2010
Local protein kinase A action proceeds through intact holoenzymes journal June 2017
High expression of XYL2 coding for xylitol dehydrogenase is necessary for efficient xylose fermentation by engineered Saccharomyces cerevisiae journal July 2012
The Yeast Ras/Cyclic AMP Pathway Induces Invasive Growth by Suppressing the Cellular Stress Response journal November 1999
The Molecular Chaperone Sse1 and the Growth Control Protein Kinase Sch9 Collaborate to Regulate Protein Kinase A Activity in Saccharomyces cerevisiae journal May 2005
Construction of fast xylose-fermenting yeast based on industrial ethanol-producing diploid Saccharomyces cerevisiae by rational design and adaptive evolution journal January 2013
Novel sensing mechanisms and targets for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae journal September 1999
Glucose-Induced Stimulation of the Ras-cAMP Pathway in Yeast Leads to Multiple Phosphorylations and Activation of 6-Phosphofructo-2-kinase journal May 2003
Engineering and Two-Stage Evolution of a Lignocellulosic Hydrolysate-Tolerant Saccharomyces cerevisiae Strain for Anaerobic Fermentation of Xylose from AFEX Pretreated Corn Stover journal September 2014
Molecular mechanisms of feedback inhibition of protein kinase A on intracellular cAMP accumulation journal August 2012
A molecular barcoded yeast ORF library enables mode-of-action analysis of bioactive compounds journal April 2009
Response of yeast cells to high glucose involves molecular and physiological differences when compared to other osmostress conditions journal June 2015
Rewiring yeast osmostress signalling through the MAPK network reveals essential and non-essential roles of Hog1 in osmoadaptation journal April 2014
Structure and function of a transcriptional network activated by the MAPK Hog1 journal October 2008
Directed Evolution Reveals Unexpected Epistatic Interactions That Alter Metabolic Regulation and Enable Anaerobic Xylose Use by Saccharomyces cerevisiae journal October 2016
Development of a D-xylose fermenting and inhibitor tolerant industrial Saccharomyces cerevisiae strain with high performance in lignocellulose hydrolysates using metabolic and evolutionary engineering journal January 2013
Rewired cellular signaling coordinates sugar and hypoxic responses for anaerobic xylose fermentation in yeast journal March 2019
Modifying Yeast Tolerance to Inhibitory Conditions of Ethanol Production Processes journal November 2015

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