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

Dataset · Thu Feb 16 00:00:00 EST 2023

DOI:https://doi.org/10.13012/B2IDB-6723545_V1· OSTI ID:3014474

Wu, Zong-Yen ^[1]; Sun, Wan ^[2]; Shen, Yihui ^[3]; Pratas, Jimmy ^[3]; Suthers, Patrick F. ^[4]; Hsieh, Ping-Hung ^[5]; Dwaraknath, Sudharsan ^[5]; Rabinowitz, Joshua D. ^[3]; Maranas, Costas D. ^[4]; Shao, Zengyi ^[6]; Yoshikuni, Yasuo ^[7]

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1. 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)
2. Interdepartmental Microbiology Program, Iowa State University, Ames, IA, 50011-1027, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
3. 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)
4. 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
5. 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)
6. 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)
7. 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

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.

• Research Organization: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign
• Sponsoring Organization: U.S. Department of Energy (DOE)
• DOE Contract Number: SC0018420
• OSTI ID: 3014474
• Country of Publication: United States
• Language: English

Related Subjects

Conversion
Metabolomics