L–malic acid production from xylose by engineered Saccharomyces cerevisiae

Kang, Nam Kyu; Lee, Jae Won; Ort, Donald R.; Jin, Yong‐Su

doi:10.1002/biot.202000431

L–malic acid production from xylose by engineered Saccharomyces cerevisiae

Journal Article · Fri Aug 13 00:00:00 EDT 2021 · Biotechnology Journal

DOI:https://doi.org/10.1002/biot.202000431· OSTI ID:1834397

Kang, Nam Kyu ^[1]; Lee, Jae Won ^[2]; Ort, Donald R. ^[2]; ^[2]

Univ. of Illinois at Urbana-Champaign, IL (United States); CABBI
Univ. of Illinois at Urbana-Champaign, IL (United States)

L-malic acid is widely used in the food, chemical, and pharmaceutical industries. Here, we report on production of malic acid from xylose, the second most abundant sugar in lignocellulosic hydrolysates, by engineered Saccharomyces cerevisiae. To enable malic acid production in a xylose-assimilating S. cerevisiae, we overexpressed PYC1 and PYC2, coding for pyruvate carboxylases, a truncated MDH3 coding for malate dehydrogenase, and SpMAE1, coding for a Schizosaccharomyces pombe malate transporter. Additionally, both the ethanol and glycerol-producing pathways were blocked to enhance malic acid production. The resulting strain produced malic acid from both glucose and xylose, but it produced much higher titers of malic acid from xylose than glucose. Interestingly, the engineered strain had higher malic acid yield from lower concentrations (10 g L^–1) of xylose, with no ethanol production, than from higher xylose concentrations (20 and 40 g L^–1). As such, a fed-batch culture maintaining xylose concentrations at low levels was conducted and 61.2 g L^–1 of malic acid was produced, with a productivity of 0.32 g L^–1 h. Furthermore, these results represent successful engineering of S. cerevisiae for the production of malic acid from xylose, confirming that that xylose offers the efficient production of various biofuels and chemicals by engineered S. cerevisiae.

View Accepted Manuscript (DOE)

Research Organization:: CABBI, Urbana, IL (United States)

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

Grant/Contract Number:: SC0018420

OSTI ID:: 1834397

Journal Information:: Biotechnology Journal, Journal Name: Biotechnology Journal Journal Issue: 3 Vol. 17; ISSN 1860-6768

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

References (45)

HeterologousHIS3 Marker and GFP Reporter Modules for PCR-Targeting inSaccharomyces cerevisiae Wach, Achim; Brachat, Arndt; Alberti-Segui, Christina Yeast, Vol. 13, Issue 11 https://doi.org/10.1002/(SICI)1097-0061(19970915)13:11<1065::AID-YEA159>3.0.CO;2-K	journal	September 1997
Fermentative capacity in high-cell-density fed-batch cultures of baker's yeast van Hoek, Pim; de Hulster, Erik; van Dijken, Johannes P. Biotechnology and Bioengineering, Vol. 68, Issue 5 https://doi.org/10.1002/(SICI)1097-0290(20000605)68:5<517::AID-BIT5>3.0.CO;2-O	journal	June 2000
Advances in metabolic engineering of yeast Saccharomyces cerevisiae for production of chemicals Borodina, Irina; Nielsen, Jens Biotechnology Journal, Vol. 9, Issue 5 https://doi.org/10.1002/biot.201300445	journal	February 2014
Rapid and marker-free refactoring of xylose-fermenting yeast strains with Cas9/CRISPR: Rapid and Marker-Free Cas9/CRISPR Refactoring Tsai, Ching-Sung; Kong, In Iok; Lesmana, Anastashia Biotechnology and Bioengineering, Vol. 112, Issue 11 https://doi.org/10.1002/bit.25632	journal	June 2015
Xylose assimilation enhances the production of isobutanol in engineered Saccharomyces cerevisiae Lane, Stephan; Zhang, Yanfei; Yun, Eun Ju Biotechnology and Bioengineering, Vol. 117, Issue 2 https://doi.org/10.1002/bit.27202	journal	November 2019
High‐level β‐carotene production from xylose by engineered Saccharomyces cerevisiae without overexpression of a truncated HMG1 (t HMG1 ) Sun, Liang; Atkinson, Christine A.; Lee, Ye‐Gi Biotechnology and Bioengineering, Vol. 117, Issue 11 https://doi.org/10.1002/bit.27508	journal	August 2020
Organic acids: old metabolites, new themes Goldberg, Israel; Rokem, J. Stefan; Pines, Ophry Journal of Chemical Technology & Biotechnology, Vol. 81, Issue 10 https://doi.org/10.1002/jctb.1590	journal	January 2006
Genome-wide identification of genes involved in tolerance to various environmental stresses inSaccharomyces cerevisiae Auesukaree, C.; Damnernsawad, A.; Kruatrachue, M. Journal of Applied Genetics, Vol. 50, Issue 3 https://doi.org/10.1007/BF03195688	journal	September 2009
Double mutation of the PDC1 and ADH1 genes improves lactate production in the yeast Saccharomyces cerevisiae expressing the bovine lactate dehydrogenase gene Tokuhiro, Kenro; Ishida, Nobuhiro; Nagamori, Eiji Applied Microbiology and Biotechnology, Vol. 82, Issue 5 https://doi.org/10.1007/s00253-008-1831-5	journal	April 2009
Engineering rTCA pathway and C4-dicarboxylate transporter for l-malic acid production Chen, Xiulai; Wang, Yuancai; Dong, Xiaoxiang Applied Microbiology and Biotechnology, Vol. 101, Issue 10 https://doi.org/10.1007/s00253-017-8141-8	journal	February 2017
Effect of yeast fumarase gene (FUM1) disruption on production of malic, fumaric and succinic acids in sake mash Magarifuchi, Tetsuro; Goto, Kuniyasu; Iimura, Yuzuru Journal of Fermentation and Bioengineering, Vol. 80, Issue 4 https://doi.org/10.1016/0922-338X(95)94204-5	journal	January 1995
Effect of NAD+-dependent isocitrate dehydrogenase gene (IDH1, IDH2) disruption of sake yeast on organic acid composition in sake mash Asano, Tadao; Kurose, Naotaka; Hiraoka, Nobutsugu Journal of Bioscience and Bioengineering, Vol. 88, Issue 3 https://doi.org/10.1016/S1389-1723(00)80006-7	journal	January 1999
Isolation of sake yeast strains possessing various levels of succinate- and/or malate-producing abilities by gene disruption or mutation Arikawa, Yukihiko; Kobayashi, Misato; Kodaira, Ritsuko Journal of Bioscience and Bioengineering, Vol. 87, Issue 3 https://doi.org/10.1016/S1389-1723(99)80041-3	journal	January 1999
Fumaric acid production in Saccharomyces cerevisiae by simultaneous use of oxidative and reductive routes Xu, Guoqiang; Chen, Xiulai; Liu, Liming Bioresource Technology, Vol. 148 https://doi.org/10.1016/j.biortech.2013.08.115	journal	November 2013
Construction of reductive pathway in Saccharomyces cerevisiae for effective succinic acid fermentation at low pH value Yan, Daojiang; Wang, Caixia; Zhou, Jiemin Bioresource Technology, Vol. 156 https://doi.org/10.1016/j.biortech.2014.01.053	journal	March 2014
Protein and metabolic engineering for the production of organic acids Liu, Jingjing; Li, Jianghua; Shin, Hyun-dong Bioresource Technology, Vol. 239 https://doi.org/10.1016/j.biortech.2017.04.052	journal	September 2017
Value-added biotransformation of cellulosic sugars by engineered Saccharomyces cerevisiae Lane, Stephan; Dong, Jia; Jin, Yong-Su Bioresource Technology, Vol. 260 https://doi.org/10.1016/j.biortech.2018.04.013	journal	July 2018
Strain engineering of Saccharomyces cerevisiae for enhanced xylose metabolism Kim, Soo Rin; Park, Yong-Cheol; Jin, Yong-Su Biotechnology Advances, Vol. 31, Issue 6 https://doi.org/10.1016/j.biotechadv.2013.03.004	journal	November 2013
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
Production of succinic acid by metabolically engineered microorganisms Ahn, Jung Ho; Jang, Yu-Sin; Lee, Sang Yup Current Opinion in Biotechnology, Vol. 42 https://doi.org/10.1016/j.copbio.2016.02.034	journal	December 2016
Genetic engineering to enhance the Ehrlich pathway and alter carbon flux for increased isobutanol production from glucose by Saccharomyces cerevisiae Kondo, Takashi; Tezuka, Hironori; Ishii, Jun Journal of Biotechnology, Vol. 159, Issue 1-2 https://doi.org/10.1016/j.jbiotec.2012.01.022	journal	May 2012
Innovative Tools and Strategies for Optimizing Yeast Cell Factories Guirimand, Gregory; Kulagina, Natalja; Papon, Nicolas Trends in Biotechnology, Vol. 39, Issue 5 https://doi.org/10.1016/j.tibtech.2020.08.010	journal	May 2021
Agriculture: Beyond food versus fuel Graham-Rowe, Duncan Nature, Vol. 474, Issue 7352 https://doi.org/10.1038/474S06a	journal	June 2011
Opportunities and challenges for a sustainable energy future Chu, Steven; Majumdar, Arun Nature, Vol. 488, Issue 7411, p. 294-303 https://doi.org/10.1038/nature11475	journal	August 2012
Enzymatic assembly of DNA molecules up to several hundred kilobases Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan Nature Methods, Vol. 6, Issue 5, p. 343-345 https://doi.org/10.1038/nmeth.1318	journal	April 2009
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
A comprehensive metabolic map for production of bio-based chemicals Lee, Sang Yup; Kim, Hyun Uk; Chae, Tong Un Nature Catalysis, Vol. 2, Issue 1 https://doi.org/10.1038/s41929-018-0212-4	journal	January 2019
Engineering energetically efficient transport of dicarboxylic acids in yeast Saccharomyces cerevisiae Darbani, Behrooz; Stovicek, Vratislav; van der Hoek, Steven Axel Proceedings of the National Academy of Sciences, Vol. 116, Issue 39 https://doi.org/10.1073/pnas.1900287116	journal	August 2019
C4-dicarboxylic acid production by overexpressing the reductive TCA pathway Zhang, Ting; Ge, Chengyu; Deng, Li FEMS Microbiology Letters, Vol. 362, Issue 9 https://doi.org/10.1093/femsle/fnv052	journal	April 2015
Assessing the effect of d-xylose on the sugar signaling pathways of Saccharomyces cerevisiae in strains engineered for xylose transport and assimilation Osiro, Karen O.; Brink, Daniel P.; Borgström, Celina FEMS Yeast Research, Vol. 18, Issue 1 https://doi.org/10.1093/femsyr/fox096	journal	January 2018
The Crabtree Effect: A Regulatory System in Yeast De Deken, R. H. Journal of General Microbiology, Vol. 44, Issue 2 https://doi.org/10.1099/00221287-44-2-149	journal	August 1966
The alcohol dehydrogenases of Saccharomyces cerevisiae: a comprehensive review de Smidt, Olga; du Preez, James C.; Albertyn, Jacobus FEMS Yeast Research, Vol. 8, Issue 7, p. 967-978 https://doi.org/10.1111/j.1567-1364.2008.00387.x	journal	November 2008
Growth requirements of pyruvate-decarboxylase-negative Saccharomyces cerevisiae Flikweert, Marcel T.; Swaaf, Martin; Dijken, Johannes P. FEMS Microbiology Letters, Vol. 174, Issue 1 https://doi.org/10.1111/j.1574-6968.1999.tb13551.x	journal	May 1999
Advanced Technology Paths to Global Climate Stability: Energy for a Greenhouse Planet Hoffert, M. I.; Caldeira, K.; Benford, G. Science, Vol. 298, Issue 5595 https://doi.org/10.1126/science.1072357	journal	November 2002
Domain Architecture of Pyruvate Carboxylase, a Biotin-Dependent Multifunctional Enzyme Maurice, M. St.; Reinhardt, L.; Surinya, K. H. Science, Vol. 317, Issue 5841 https://doi.org/10.1126/science.1144504	journal	August 2007
Construction of a Quadruple Auxotrophic Mutant of an Industrial Polyploid Saccharomyces cerevisiae Strain by Using RNA-Guided Cas9 Nuclease Zhang, Guo-Chang; Kong, In Iok; Kim, Heejin Applied and Environmental Microbiology, Vol. 80, Issue 24 https://doi.org/10.1128/AEM.02310-14	journal	October 2014
Malic Acid Production by Saccharomyces cerevisiae: Engineering of Pyruvate Carboxylation, Oxaloacetate Reduction, and Malate Export Zelle, R. M.; de Hulster, E.; van Winden, W. A. Applied and Environmental Microbiology, Vol. 74, Issue 9 https://doi.org/10.1128/AEM.02591-07	journal	March 2008
Saccharomyces cerevisiae Engineered for Xylose Metabolism Exhibits a Respiratory Response Jin, Y.-S.; Laplaza, J. M.; Jeffries, T. W. Applied and Environmental Microbiology, Vol. 70, Issue 11, p. 6816-6825 https://doi.org/10.1128/AEM.70.11.6816-6825.2004	journal	November 2004
Cellulosic Biofuels Carroll, Andrew; Somerville, Chris Annual Review of Plant Biology, Vol. 60, Issue 1 https://doi.org/10.1146/annurev.arplant.043008.092125	journal	June 2009
Scheffersomyces stipitis: a comparative systems biology study with the Crabtree positive yeast Saccharomyces cerevisiae Papini, Marta; Nookaew, Intawat; Uhlén, Mathias Microbial Cell Factories, Vol. 11, Issue 1 https://doi.org/10.1186/1475-2859-11-136	journal	January 2012
Transcription analysis of recombinant industrial and laboratory Saccharomyces cerevisiae strains reveals the molecular basis for fermentation of glucose and xylose Matsushika, Akinori; Goshima, Tetsuya; Hoshino, Tamotsu Microbial Cell Factories, Vol. 13, Issue 1 https://doi.org/10.1186/1475-2859-13-16	journal	January 2014
Production of fuels and chemicals from xylose by engineered Saccharomyces cerevisiae: a review and perspective Kwak, Suryang; Jin, Yong-Su Microbial Cell Factories, Vol. 16, Issue 1 https://doi.org/10.1186/s12934-017-0694-9	journal	May 2017
Production of a human milk oligosaccharide 2′-fucosyllactose by metabolically engineered Saccharomyces cerevisiae Yu, Sora; Liu, Jing-Jing; Yun, Eun Ju Microbial Cell Factories, Vol. 17, Issue 1 https://doi.org/10.1186/s12934-018-0947-2	journal	June 2018
Production of 2,3-butanediol from glucose and cassava hydrolysates by metabolically engineered industrial polyploid Saccharomyces cerevisiae Lee, Ye-Gi; Seo, Jin-Ho Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1545-1	journal	August 2019
An evolutionary perspective on the Crabtree effect Pfeiffer, Thomas; Morley, Annabel Frontiers in Molecular Biosciences, Vol. 1 https://doi.org/10.3389/fmolb.2014.00017	journal	October 2014

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