Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae

Kwak, Suryang; Yun, Eun Ju; Lane, Stephan; Oh, Eun Joong; Kim, Kyoung Heon; Jin, Yong‐Su

doi:10.1002/biot.201900173

Title: Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae

Journal Article · 29 August 2019 · Biotechnology Journal

DOI: https://doi.org/10.1002/biot.201900173 · OSTI ID:1559936

^[1]; Yun, Eun Ju ^[2]; Lane, Stephan ^[1]; Oh, Eun Joong ^[1]; Kim, Kyoung Heon ^[3];

^[1]

Univ. of Illinois, Urbana-Champaign, IL (United States). Dept. of Food Science and Human Nutrition and Carl R. Woese Institute for Genomic Biology
Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Institute for Genomic Biology; Korea Univ., Seoul (Korea). Dept. of Biotechnology
Korea Univ., Seoul (Korea). Dept. of Biotechnology

Abstract Sufficient supply of reduced nicotinamide adenine dinucleotide phosphate (NADPH) is a prerequisite of the overproduction of isoprenoids and related bioproducts in Saccharomyces cerevisiae . Although S. cerevisiae highly depends on the oxidative pentose phosphate (PP) pathway to produce NADPH, its metabolic flux toward the oxidative PP pathway is limited due to the rigid glycolysis flux. To maximize NADPH supply for the isoprenoid production in yeast, upper glycolytic metabolic fluxes are reduced by introducing mutations into phosphofructokinase (PFK) along with overexpression of ZWF1 encoding glucose‐6‐phosphate (G6P) dehydrogenase. The PFK mutations (Pfk1 S724D and Pfk2 S718D) result in less glycerol production and more accumulation of G6P, which is a gateway metabolite toward the oxidative PP pathway. When combined with the PFK mutations, overexpression of ZWF1 caused substantial increases of [NADPH]/[NADP ⁺ ] ratios whereas the effect of ZWF1 overexpression alone in the wild‐type strain is not noticeable. Also, the introduction of ZWF1 overexpression and the PFK mutations into engineered yeast overexpressing acetyl‐CoA C‐acetyltransferase ( ERG10 ), truncated HMG‐CoA reductase isozyme 1 ( tHMG1 ), and amorphadiene synthase ( ADS ) leads to a titer of 497 mg L ^–1 of amorphadiene (3.7‐fold over the parental strain). These results suggest that perturbation of upper glycolytic fluxes, in addition to ZWF1 overexpression, is necessary for efficient NADPH supply through the oxidative PP pathway and enhanced production of isoprenoids by engineered S. cerevisiae .

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Research Organization:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

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

Grant/Contract Number:: SC0018420; DE‐SC0018420

OSTI ID:: 1559936

Alternate ID(s):: OSTI ID: 1562316

Journal Information:: Biotechnology Journal, Vol. none, Issue none; ISSN 1860-6768

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 18 works

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References (58)

Manipulation of malic enzyme in Saccharomyces cerevisiae for increasing NADPH production capacity aerobically in different cellular compartments Moreira dos Santos, Margarida; Raghevendran, Vijayendran; Kötter, Peter Metabolic Engineering, Vol. 6, Issue 4 https://doi.org/10.1016/j.ymben.2004.06.002	journal	October 2004
Connecting extracellular metabolomic measurements to intracellular flux states in yeast Mo, Monica L.; Palsson, Bernhard Ø; Herrgård, Markus J. BMC Systems Biology, Vol. 3, Issue 1 https://doi.org/10.1186/1752-0509-3-37	journal	January 2009
Production and quantification of sesquiterpenes in Saccharomyces cerevisiae, including extraction, detection and quantification of terpene products and key related metabolites Rodriguez, Sarah; Kirby, James; Denby, Charles M. Nature Protocols, Vol. 9, Issue 8 https://doi.org/10.1038/nprot.2014.132	journal	July 2014
Reprogramming Yeast Metabolism from Alcoholic Fermentation to Lipogenesis Yu, Tao; Zhou, Yongjin J.; Huang, Mingtao Cell, Vol. 174, Issue 6 https://doi.org/10.1016/j.cell.2018.07.013	journal	September 2018
Redirection of flux through the FPP branch-point in Saccharomyces cerevisiae by down-regulating squalene synthase Paradise, Eric M.; Kirby, James; Chan, Rossana Biotechnology and Bioengineering, Vol. 100, Issue 2 https://doi.org/10.1002/bit.21766	journal	June 2008
Total biosynthesis of hydrocortisone from a simple carbon source in yeast Szczebara, Florence Ménard; Chandelier, Cathy; Villeret, Coralie Nature Biotechnology, Vol. 21, Issue 2 https://doi.org/10.1038/nbt775	journal	January 2003
Combinatorial engineering of mevalonate pathway for improved amorpha-4,11-diene production in budding yeast: Combinatorial Engineering of Mevalonate Pathway Yuan, Jifeng; Ching, Chi Bun Biotechnology and Bioengineering, Vol. 111, Issue 3 https://doi.org/10.1002/bit.25123	journal	October 2013
Metabolic engineering of Saccharomyces cerevisiae for production of ginsenosides Dai, Zhubo; Liu, Yi; Zhang, Xianan Metabolic Engineering, Vol. 20 https://doi.org/10.1016/j.ymben.2013.10.004	journal	November 2013
FastCloning: a highly simplified, purification-free, sequence- and ligation-independent PCR cloning method Li, Chaokun; Wen, Aiyun; Shen, Benchang BMC Biotechnology, Vol. 11, Issue 1 https://doi.org/10.1186/1472-6750-11-92	journal	January 2011
Glutathione reductase directly mediates the stimulation of yeast glucose-6-phosphate dehydrogenase by GSSG Llobell, A.; Lopez-Ruiz, A.; Peinado, J. Biochemical Journal, Vol. 249, Issue 1 https://doi.org/10.1042/bj2490293	journal	January 1988
Reduced methylation of PFKFB3 in cancer cells shunts glucose towards the pentose phosphate pathway Yamamoto, Takehiro; Takano, Naoharu; Ishiwata, Kyoko Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms4480	journal	March 2014
Engineering cellular redox balance in Saccharomyces cerevisiae for improved production of L-lactic acid : Engineering Redox Balance for Lactic Acid Production Lee, Ju Young; Kang, Chang Duk; Lee, Seung Hyun Biotechnology and Bioengineering, Vol. 112, Issue 4 https://doi.org/10.1002/bit.25488	journal	January 2015
Cloning of a second gene encoding 6-phosphofructo-2-kinase in yeast, and characterization of mutant strains without fructose-2,6-bisphosphate Bolesm, Eckhard; Göhlmann, Hinrich W. H.; Zimmermann, Friedrich K. Molecular Microbiology, Vol. 20, Issue 1 https://doi.org/10.1111/j.1365-2958.1996.tb02489.x	journal	April 1996
Microbial engineering for the production of advanced biofuels Peralta-Yahya, Pamela P.; Zhang, Fuzhong; del Cardayre, Stephen B. Nature, Vol. 488, Issue 7411 https://doi.org/10.1038/nature11478	journal	August 2012
Engineering redox balance through cofactor systems Chen, Xiulai; Li, Shubo; Liu, Liming Trends in Biotechnology, Vol. 32, Issue 6 https://doi.org/10.1016/j.tibtech.2014.04.003	journal	June 2014
A Dominant Mutation That Alters the Regulation of INO1 Expression in Saccharomyces cerevisiae Hosaka, Kohei; Nikawa, Jun-ichi; Kodaki, Tsutomu The Journal of Biochemistry, Vol. 111, Issue 3 https://doi.org/10.1093/oxfordjournals.jbchem.a123761	journal	March 1992
High-level semi-synthetic production of the potent antimalarial artemisinin Paddon, C. J.; Westfall, P. J.; Pitera, D. J. Nature, Vol. 496, Issue 7446 https://doi.org/10.1038/nature12051	journal	April 2013
Profiling of Cytosolic and Peroxisomal Acetyl-CoA Metabolism in Saccharomyces cerevisiae Chen, Yun; Siewers, Verena; Nielsen, Jens PLoS ONE, Vol. 7, Issue 8 https://doi.org/10.1371/journal.pone.0042475	journal	August 2012
High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method Gietz, R. Daniel; Schiestl, Robert H. Nature Protocols, Vol. 2, Issue 1 https://doi.org/10.1038/nprot.2007.13	journal	January 2007
COBRApy: COnstraints-Based Reconstruction and Analysis for Python Ebrahim, Ali; Lerman, Joshua A.; Palsson, Bernhard O. BMC Systems Biology, Vol. 7, Issue 1 https://doi.org/10.1186/1752-0509-7-74	journal	January 2013
Lost in Transition: Start-Up of Glycolysis Yields Subpopulations of Nongrowing Cells van Heerden, J. H.; Wortel, M. T.; Bruggeman, F. J. Science, Vol. 343, Issue 6174 https://doi.org/10.1126/science.1245114	journal	January 2014
Dual Activators of the Sterol Biosynthetic Pathway of Saccharomyces cerevisiae: Similar Activation/Regulatory Domains but Different Response Mechanisms Davies, B. S. J.; Wang, H. S.; Rine, J. Molecular and Cellular Biology, Vol. 25, Issue 16 https://doi.org/10.1128/MCB.25.16.7375-7385.2005	journal	July 2005
Observations on squalene accumulation in Saccharomyces cerevisiae due to the manipulation of HMG2 and ERG6: Observations on squalene accumulation capacity in yeast Mantzouridou, Fani; Tsimidou, Maria Z. FEMS Yeast Research, Vol. 10, Issue 6 https://doi.org/10.1111/j.1567-1364.2010.00645.x	journal	August 2010
A Yeast Phosphofructokinase Insensitive to the Allosteric Activator Fructose 2,6-Bisphosphate: GLYCOLYSIS/METABOLIC REGULATION/ALLOSTERIC CONTROL Heinisch, Jürgen J.; Boles, Eckhard; Timpel, Claudia Journal of Biological Chemistry, Vol. 271, Issue 27 https://doi.org/10.1074/jbc.271.27.15928	journal	July 1996
Advancing Metabolic Engineering of Saccharomyces cerevisiae Using the CRISPR/Cas System Lian, Jiazhang; HamediRad, Mohammad; Zhao, Huimin Biotechnology Journal, Vol. 13, Issue 9 https://doi.org/10.1002/biot.201700601	journal	April 2018
Production of amorphadiene in yeast, and its conversion to dihydroartemisinic acid, precursor to the antimalarial agent artemisinin Westfall, P. J.; Pitera, D. J.; Lenihan, J. R. Proceedings of the National Academy of Sciences, Vol. 109, Issue 3, p. E111-E118 https://doi.org/10.1073/pnas.1110740109	journal	January 2012
Enzymatic Activity Assays in Yeast Cell Extracts Çağlayan, Melike; Wilson, Samuel BIO-PROTOCOL, Vol. 4, Issue 23 https://doi.org/10.21769/BioProtoc.1312	journal	January 2014
Rewriting yeast central carbon metabolism for industrial isoprenoid production Meadows, Adam L.; Hawkins, Kristy M.; Tsegaye, Yoseph Nature, Vol. 537, Issue 7622 https://doi.org/10.1038/nature19769	journal	September 2016
Transcriptional control of the yeast acetyl‐CoA synthetase gene, ACS1 , by the positive regulators CAT8 and ADR1 and the pleiotropic repressor UME6 Kratzer, Steffen; Schüller, Hans‐Joachim Molecular Microbiology, Vol. 26, Issue 4 https://doi.org/10.1046/j.1365-2958.1997.5611937.x	journal	November 1997
Metabolic-Flux Profiling of the Yeasts Saccharomyces cerevisiae and Pichia stipitis Fiaux, Jocelyne; Çakar, Z. Petek; Sonderegger, Marco Eukaryotic Cell, Vol. 2, Issue 1 https://doi.org/10.1128/EC.2.1.170-180.2003	journal	February 2003
Stoichiometry and compartmentation of NADH metabolism in Saccharomyces cerevisiae Bakker, Barbara M.; Overkamp, Karin M.; van Maris, Antonius J. A. FEMS Microbiology Reviews, Vol. 25, Issue 1, p. 15-37 https://doi.org/10.1111/j.1574-6976.2001.tb00570.x	journal	January 2001
Fps1, a yeast member of the MIP family of channel proteins, is a facilitator for glycerol uptake and efflux and is inactive under osmotic stress. Luyten, K.; Albertyn, J.; Skibbe, W. F. The EMBO Journal, Vol. 14, Issue 7 https://doi.org/10.1002/j.1460-2075.1995.tb07122.x	journal	April 1995
Genome-Scale Reconstruction of the Saccharomyces cerevisiae Metabolic Network Forster, J. Genome Research, Vol. 13, Issue 2 https://doi.org/10.1101/gr.234503	journal	February 2003
Dynamic control of ERG9 expression for improved amorpha-4,11-diene production in Saccharomyces cerevisiae Yuan, Jifeng; Ching, Chi-Bun Microbial Cell Factories, Vol. 14, Issue 1 https://doi.org/10.1186/s12934-015-0220-x	journal	March 2015
Studies on the mechanism of the glucose-induced cAMP signal in glycolysis and glucose repression mutants of the yeast Saccharomyces cerevisiae Beullens, Monique; Mbonyi, Kaishusha; Geerts, Lieve European Journal of Biochemistry, Vol. 172, Issue 1 https://doi.org/10.1111/j.1432-1033.1988.tb13877.x	journal	February 1988
Production of the antimalarial drug precursor artemisinic acid in engineered yeast Ro, Dae-Kyun; Paradise, Eric M.; Ouellet, Mario Nature, Vol. 440, Issue 7086, p. 940-943 https://doi.org/10.1038/nature04640	journal	April 2006
Systematic Identification of Conserved Metabolites in GC/MS Data for Metabolomics and Biomarker Discovery Styczynski, Mark P.; Moxley, Joel F.; Tong, Lily V. Analytical Chemistry, Vol. 79, Issue 3 https://doi.org/10.1021/ac0614846	journal	February 2007
Glycerol and arabitol production by an intergeneric hybrid, PB2, obtained by protoplast fusion between Saccharomyces cerevisiae and Torulaspora delbrueckii M., Lucca; J., Spencer; Figueroa, L. de Applied Microbiology and Biotechnology, Vol. 59, Issue 4-5 https://doi.org/10.1007/s00253-002-1025-5	journal	January 2002
It Is All about Metabolic Fluxes Nielsen, J. Journal of Bacteriology, Vol. 185, Issue 24 https://doi.org/10.1128/JB.185.24.7031-7035.2003	journal	December 2003
Temporal system‐level organization of the switch from glycolytic to gluconeogenic operation in yeast Zampar, Guillermo G.; Kümmel, Anne; Ewald, Jennifer Molecular Systems Biology, Vol. 9, Issue 1 https://doi.org/10.1038/msb.2013.11	journal	January 2013
Enhancing the flux of D-glucose to the pentose phosphate pathway in Saccharomyces cerevisiae for the production of D-ribose and ribitol Toivari, Mervi H.; Maaheimo, Hannu; Penttilä, Merja Applied Microbiology and Biotechnology, Vol. 85, Issue 3 https://doi.org/10.1007/s00253-009-2184-4	journal	August 2009
Sources of NADPH in Yeast Vary with Carbon Source Minard, Karyl I.; McAlister-Henn, Lee Journal of Biological Chemistry, Vol. 280, Issue 48 https://doi.org/10.1074/jbc.M509461200	journal	September 2005
An integrated method for spectrum extraction and compound identification from gas chromatography/mass spectrometry data Stein, S. E. Journal of the American Society for Mass Spectrometry, Vol. 10, Issue 8 https://doi.org/10.1016/S1044-0305(99)00047-1	journal	August 1999
Production of biofuels and chemicals from xylose using native and engineered yeast strains Kwak, Suryang; Jo, Jung Hyun; Yun, Eun Ju Biotechnology Advances, Vol. 37, Issue 2 https://doi.org/10.1016/j.biotechadv.2018.12.003	journal	March 2019
GMD@CSB.DB: the Golm Metabolome Database Kopka, J.; Schauer, N.; Krueger, S. Bioinformatics, Vol. 21, Issue 8 https://doi.org/10.1093/bioinformatics/bti236	journal	December 2004
A Novel Strategy for Selection and Validation of Reference Genes in Dynamic Multidimensional Experimental Design in Yeast Cankorur-Cetinkaya, Ayca; Dereli, Elif; Eraslan, Serpil PLoS ONE, Vol. 7, Issue 6 https://doi.org/10.1371/journal.pone.0038351	journal	June 2012
Production of isoprenoid pharmaceuticals by engineered microbes Chang, Michelle C. Y.; Keasling, Jay D. Nature Chemical Biology, Vol. 2, Issue 12 https://doi.org/10.1038/nchembio836	journal	November 2006
Combined metabolic engineering of precursor and co-factor supply to increase α-santalene production by Saccharomyces cerevisiae Scalcinati, Gionata; Partow, Siavash; Siewers, Verena Microbial Cell Factories, Vol. 11, Issue 1 https://doi.org/10.1186/1475-2859-11-117	journal	January 2012
Metabolic engineering delivers next-generation biofuels Keasling, Jay D.; Chou, Howard Nature Biotechnology, Vol. 26, Issue 3 https://doi.org/10.1038/nbt0308-298	journal	March 2008
Metabolic Impact of Redox Cofactor Perturbations on the Formation of Aroma Compounds in Saccharomyces cerevisiae Bloem, Audrey; Sanchez, Isabelle; Dequin, Sylvie Applied and Environmental Microbiology, Vol. 82, Issue 1 https://doi.org/10.1128/AEM.02429-15	journal	October 2015
Evaluation and Optimization of Metabolome Sample Preparation Methods for Saccharomyces cerevisiae Kim, Sooah; Lee, Do Yup; Wohlgemuth, Gert Analytical Chemistry, Vol. 85, Issue 4 https://doi.org/10.1021/ac302881e	journal	January 2013
Enhanced isoprenoid production from xylose by engineered Saccharomyces cerevisiae : Enhanced Yeast Isoprenoid Production from Xylose Kwak, Suryang; Kim, Soo Rin; Xu, Haiqing Biotechnology and Bioengineering, Vol. 114, Issue 11 https://doi.org/10.1002/bit.26369	journal	July 2017
Production of the Carotenoids Lycopene, β-Carotene, and Astaxanthin in the Food Yeast Candida utilis Miura, Yutaka; Kondo, Keiji; Saito, Toshiko Applied and Environmental Microbiology, Vol. 64, Issue 4 https://doi.org/10.1128/AEM.64.4.1226-1229.1998	journal	January 1998
Observations on squalene accumulation in Saccharomyces cerevisiae due to the manipulation of HMG2 and ERG6: Erratum Mantzouridou, Fani; Tsimidou, Maria Z. FEMS Yeast Research, Vol. 11, Issue 2 https://doi.org/10.1111/j.1567-1364.2011.00717.x	journal	January 2011
Engineering of Saccharomyces cerevisiae for anthranilate and methyl anthranilate production Kuivanen, Joosu; Kannisto, Matti; Mojzita, Dominik Microbial Cell Factories, Vol. 20, Issue 1 https://doi.org/10.1186/s12934-021-01532-3	journal	February 2021
Temporal system-level organization of the switch from glycolytic to gluconeogenic operation in yeast Zampar, Guillermo G.; Kümmel, Anne; Ewald, Jennifer European Molecular Biology Organization https://doi.org/10.5167/uzh-79184	text	January 2013
GMD@CSB.DB: The Golm Metabolome Database Kopka, Joachim; Schauer, Nicolas; Krueger, Stephan ETH Zurich https://doi.org/10.3929/ethz-b-000033179	text	January 2005
Temporal system-level organization of the switch from glycolytic to gluconeogenic operation in yeast Zampar, Guillermo G.; Kümmel, Anne; Ewald, Jennifer ETH Zurich https://doi.org/10.3929/ethz-b-000066745	text	January 2013

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59 BASIC BIOLOGICAL SCIENCES
isoprenoid
NADPH
oxidative pentose phosphate pathway
phosphofructokinase
Saccharomyces cerevisiae

Title: Redirection of the Glycolytic Flux Enhances Isoprenoid Production in Saccharomyces cerevisiae

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