ArticleApplied and environmental microbiology2011
Anaplerotic role for cytosolic malic enzyme in engineered Saccharomyces cerevisiae strains.
Article in Applied and environmental microbiology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers, 1 of them a synthesis that pooled it.
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Who cites it
21 citing papers in PubMed, 1 synthesis or guideline pooled it, 58 citations in OpenAlex.
- Pooled it
- Research on Engineering theFoods (Basel, Switzerland) · 2024Article
- Effects of Heterologous Expression of Genes Related L-Malic acid Metabolism inFoods (Basel, Switzerland) · 2024Article
- Automated in vivo enzyme engineering accelerates biocatalyst optimization.Nature communications · 2024Review
- Engineering yeast mitochondrial metabolism for 3-hydroxypropionate production.Biotechnology for biofuels and bioproducts · 2023Article
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- Awakening a latent carbon fixation cycle in Escherichia coli.Nature communications · 2020Article
- The Role of Yeasts and Lactic Acid Bacteria on the Metabolism of Organic Acids during Winemaking.Foods (Basel, Switzerland) · 2020Review
- Comprehensive Analysis ofFrontiers in bioengineering and biotechnology · 2020Article
- The emergence of adaptive laboratory evolution as an efficient tool for biological discovery and industrial biotechnology.Metabolic engineering · 2019Review
- The Role of Malic Enzyme on Promoting Total Lipid and Fatty Acid Production inFrontiers in plant science · 2018Article
- Holistic bioengineering: rewiring central metabolism for enhanced bioproduction.The Biochemical journal · 2017Review
- Mitochondrial Pyruvate Import Promotes Long-Term Survival of Antibody-Secreting Plasma Cells.Immunity · 2016Article
- Embryonic Lethality of Mitochondrial Pyruvate Carrier 1 Deficient Mouse Can Be Rescued by a Ketogenic Diet.PLoS genetics · 2016Article
- Elemental sulfur and acetate can support life of a novel strictly anaerobic haloarchaeon.The ISME journal · 2016Article
- Metabolomic profiling of the purple sulfur bacteriumMetabolomics : Official journal of the Metabolomic Society · 2014Article
- Genome duplication and mutations in ACE2 cause multicellular, fast-sedimenting phenotypes in evolved Saccharomyces cerevisiae.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Adaptive laboratory evolution -- principles and applications for biotechnology.Microbial cell factories · 2013Review
- An internal deletion in MTH1 enables growth on glucose of pyruvate-decarboxylase negative, non-fermentative Saccharomyces cerevisiae.Microbial cell factories · 2012Article
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4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Malic enzyme catalyzes the reversible oxidative decarboxylation of malate to pyruvate and CO(2). The Saccharomyces cerevisiae MAE1 gene encodes a mitochondrial malic enzyme whose proposed physiological roles are related to the oxidative, malate-decarboxylating reaction. Hitherto, the inability of pyruvate carboxylase-negative (Pyc(-)) S. cerevisiae strains to grow on glucose suggested that Mae1p cannot act as a pyruvate-carboxylating, anaplerotic enzyme. In this study, relocation of malic enzyme to the cytosol and creation of thermodynamically favorable conditions for pyruvate carboxylation by metabolic engineering, process design, and adaptive evolution, enabled malic enzyme to act as the sole anaplerotic enzyme in S. cerevisiae. The Escherichia coli NADH-dependent sfcA malic enzyme was expressed in a Pyc(-) S. cerevisiae background. When PDC2, a transcriptional regulator of pyruvate decarboxylase genes, was deleted to increase intracellular pyruvate levels and cells were grown under a CO(2) atmosphere to favor carboxylation, adaptive evolution yielded a strain that grew on glucose (specific growth rate, 0.06 ± 0.01 h(-1)). Growth of the evolved strain was enabled by a single point mutation (Asp336Gly) that switched the cofactor preference of E. coli malic enzyme from NADH to NADPH. Consistently, cytosolic relocalization of the native Mae1p, which can use both NADH and NADPH, in a pyc1,2Δ pdc2Δ strain grown under a CO(2) atmosphere, also enabled slow-growth on glucose. Although growth rates of these strains are still low, the higher ATP efficiency of carboxylation via malic enzyme, compared to the pyruvate carboxylase pathway, may contribute to metabolic engineering of S. cerevisiae for anaerobic, high-yield C(4)-dicarboxylic acid production.
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