Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate

Wu, Zong-Yen; Sun, Wan; Shen, Yihui; Pratas, Jimmy; Suthers, Patrick F.; Hsieh, Ping-Hung; Dwaraknath, Sudharsan; Rabinowitz, Joshua D.; Maranas, Costas D.; Shao, Zengyi; Yoshikuni, Yasuo

doi:10.13012/B2IDB-6723545_V1

Title: Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate

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Abstract

Methyl methacrylate (MMA) is an important petrochemical with many applications. However, its manufacture has a large environmental footprint. Combined biological and chemical synthesis (semisynthesis) may be a promising alternative to reduce both cost and environmental impact, but strains that can produce the MMA precursor (citramalate) at low pH are required. A non-conventional yeast, Issatchenkia orientalis, may prove ideal, as it can survive extremely low pH. Here, we demonstrate the engineering of I. orientalis for citramalate production. Using sequence similarity network analysis and subsequent DNA synthesis, we selected a more active citramalate synthase gene (cimA) variant for expression in I. orientalis. We then adapted a piggyBac transposon system for I. orientalis that allowed us to simultaneously explore the effects of different cimA gene copy numbers and integration locations. A batch fermentation showed the genome-integrated-cimA strains produced 2.0 g/L citramalate in 48 h and a yield of up to 7% mol citramalate/mol consumed glucose. These results demonstrate the potential of I. orientalis as a chassis for citramalate production.

Authors:

Wu, Zong-Yen; Sun, Wan; Shen, Yihui; Pratas, Jimmy; Suthers, Patrick F.; Hsieh, Ping-Hung; Dwaraknath, Sudharsan; Rabinowitz, Joshua D.; Maranas, Costas D.; Shao, Zengyi; Yoshikuni, Yasuo

Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Interdepartmental Microbiology Program, Iowa State University, Ames, IA, 50011-1027, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA; Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, 08540, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Department of Chemical Engineering, The Pennsylvania State University, University Park, PA, 16802, USA; Center for Advanced Bioenergy and Bioproducts Innovation, The Pennsylvania State University, University Park, PA, 16802, USA
US Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Interdepartmental Microbiology Program, Iowa State University, Ames, IA, 50011-1027, USA; Department of Chemical and Biological Engineering, Iowa State University, Ames, IA, 50011, USA; NSF Engineering Research Center for Biorenewable Chemicals, Iowa State University, Ames, IA, 50011, USA; Bioeconomy Institute, Iowa State University, Ames, IA, 50011, USA; The Ames Laboratory, Ames, IA, 50011, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Center for Advanced Bioenergy and Bioproducts Innovation, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Global Center for Food, Land, and Water Resources, Hokkaido University, Hokkaido, 060-8589, Japan

Publication Date:: Thu Feb 16 00:00:00 UTC 2023

DOE Contract Number:: SC0018420

Research Org.:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign

Sponsoring Org.:: U.S. Department of Energy (DOE)

Subject:: Conversion; Metabolomics

OSTI Identifier:: 3014474

DOI:: https://doi.org/10.13012/B2IDB-6723545_V1

Citation Formats


                    Wu, Zong-Yen, Sun, Wan, Shen, Yihui, Pratas, Jimmy, Suthers, Patrick F., Hsieh, Ping-Hung, Dwaraknath, Sudharsan, Rabinowitz, Joshua D., Maranas, Costas D., Shao, Zengyi, and Yoshikuni, Yasuo. Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate.  United States: N. p., 2023. 
        Web.  doi:10.13012/B2IDB-6723545_V1.

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                    Wu, Zong-Yen, Sun, Wan, Shen, Yihui, Pratas, Jimmy, Suthers, Patrick F., Hsieh, Ping-Hung, Dwaraknath, Sudharsan, Rabinowitz, Joshua D., Maranas, Costas D., Shao, Zengyi, & Yoshikuni, Yasuo. Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate.  United States.  doi:https://doi.org/10.13012/B2IDB-6723545_V1

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                    Wu, Zong-Yen, Sun, Wan, Shen, Yihui, Pratas, Jimmy, Suthers, Patrick F., Hsieh, Ping-Hung, Dwaraknath, Sudharsan, Rabinowitz, Joshua D., Maranas, Costas D., Shao, Zengyi, and Yoshikuni, Yasuo. 2023.  
"Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate".  United States.  doi:https://doi.org/10.13012/B2IDB-6723545_V1.  https://www.osti.gov/servlets/purl/3014474. Pub date:Thu Feb 16 00:00:00 UTC 2023

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@article{osti_3014474,

  title        = {Data for Metabolic Engineering of Low-pH-Tolerant Non-Model Yeast, Issatchenkia orientalis, for Production of Citramalate},

  author       = {Wu, Zong-Yen and Sun, Wan and Shen, Yihui and Pratas, Jimmy and Suthers, Patrick F. and Hsieh, Ping-Hung and Dwaraknath, Sudharsan and Rabinowitz, Joshua D. and Maranas, Costas D. and Shao, Zengyi and Yoshikuni, Yasuo},

  abstractNote = {Methyl methacrylate (MMA) is an important petrochemical with many applications. However, its manufacture has a large environmental footprint. Combined biological and chemical synthesis (semisynthesis) may be a promising alternative to reduce both cost and environmental impact, but strains that can produce the MMA precursor (citramalate) at low pH are required. A non-conventional yeast, Issatchenkia orientalis, may prove ideal, as it can survive extremely low pH. Here, we demonstrate the engineering of I. orientalis for citramalate production. Using sequence similarity network analysis and subsequent DNA synthesis, we selected a more active citramalate synthase gene (cimA) variant for expression in I. orientalis. We then adapted a piggyBac transposon system for I. orientalis that allowed us to simultaneously explore the effects of different cimA gene copy numbers and integration locations. A batch fermentation showed the genome-integrated-cimA strains produced 2.0 g/L citramalate in 48 h and a yield of up to 7% mol citramalate/mol consumed glucose. These results demonstrate the potential of I. orientalis as a chassis for citramalate production.},

  doi          = {10.13012/B2IDB-6723545_V1},

  journal      = {},

  number       = ,

  volume       = ,

  place        = {United States},

  year         = {Thu Feb 16 00:00:00 UTC 2023},

  month        = {Thu Feb 16 00:00:00 UTC 2023}

}

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DOI: https://doi.org/10.13012/B2IDB-6723545_V1

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Metabolic engineering of low-pH-tolerant non-model yeast, Issatchenkia orientalis, for production of citramalate

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