Fumaric acid overproduction in yeast mutants deficient in fumarase

Food Research Institute, Bratislava, Czechoslovakia.
FEMS Microbiology Letters (Impact Factor: 2.12). 04/1992; 70(2):101-6. DOI: 10.1016/0378-1097(92)90667-D
Source: PubMed


Abstract A nuclear mutant of Saccharomyces cerevisiae deficient in mitochondrial fumarase has been identified through the in vitro biochemical assay of enzyme activity after visual selection due to an increased acidification ability of its colonies. Cells of the fumarase-deficient mutant fermenting glucose accumulated extracellular fumaric acid. This accumulation was observed only in growing cultures and required functional mitochondrial electron transport from succinate dehydrogenase to oxygen.

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Available from: Julius Subík, Apr 13, 2014
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    • "In the second strategy, FA can be produced via an oxidative TCA cycle and the engineered strain is stable in the fermentation process. It was reported that cells of a fumarase-deficient mutant accumulated extracellular FA when fermenting glucose [11]. Similarly, a concentration of 3.62 g L–1 at a yield of 0.11 moles of succinic acid per mole of glucose was achieved for oxidative production of succinic acid in yeast by deletion of the SDH1, SDH2, IDH1 and IDP1 genes [8]. "
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    ABSTRACT: Fumaric acid (FA) is a promising biomass-derived building-block chemical. Bio-based FA production from renewable feedstock is a promising and sustainable alternative to petroleum-based chemical synthesis. Here we report on FA production by direct fermentation using metabolically engineered Saccharomyces cerevisiae with the aid of in silico analysis of a genome-scale metabolic model. First, FUM1 was selected as the target gene on the basis of extensive literature mining. Flux balance analysis (FBA) revealed that FUM1 deletion can lead to FA production and slightly lower growth of S. cerevisiae . The engineered S. cerevisiae strain obtained by deleting FUM1 can produce FA up to a concentration of 610±31 mg L<sup>–1</sup> without any apparent change in growth in fed-batch culture. FT-IR and <sup>1</sup>H and <sup>13</sup>C NMR spectra confirmed that FA was synthesized by the engineered S. cerevisiae strain. FBA identified pyruvate carboxylase as one of the factors limiting higher FA production. When the RoPYC gene was introduced, S. cerevisiae produced 1134±48 mg L<sup>–1</sup> FA. Furthermore, the final engineered S. cerevisiae strain was able to produce 1675±52 mg L<sup>–1</sup> FA in batch culture when the SFC1 gene encoding a succinate–fumarate transporter was introduced. These results demonstrate that the model shows great predictive capability for metabolic engineering. Moreover, FA production in S. cerevisiae can be efficiently developed with the aid of in silico metabolic engineering.
    PLoS ONE 12/2012; 7(12-12):e52086. DOI:10.1371/journal.pone.0052086 · 3.23 Impact Factor
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