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

Autor:	Wu ZY; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA., Sun W; Interdepartmental Microbiology Program, Iowa State University, Ames, IA, 50011-1027, USA.; DOE Center for Advanced Bioenergy and Bioproducts Innovation, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA., Shen Y; Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA.; Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, 08540, USA., Pratas J; Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA.; Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, 08540, USA., Suthers PF; 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., Hsieh PH; US Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA., Dwaraknath S; US Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA., Rabinowitz JD; Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA.; Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, 08540, USA., Maranas CD; 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., Shao Z; 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.; DOE Center for Advanced Bioenergy and Bioproducts Innovation, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA., Yoshikuni Y; 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.
Jazyk:	angličtina
Zdroj:	Metabolic engineering communications [Metab Eng Commun] 2023 Feb 16; Vol. 16, pp. e00220. Date of Electronic Publication: 2023 Feb 16 (Print Publication: 2023).
DOI:	10.1016/j.mec.2023.e00220
Abstrakt:	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. Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (© 2023 The Authors.)
Databáze:	MEDLINE
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