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Constructing recombinant Saccharomyces cerevisiae strains for malic-to-fumaric acid conversion
Annica Steyn1, Marinda Viljoen-Bloom1, Willem Heber Van Zyl1
1Department of Microbiology, Stellenbosch University, Stellenbosch 7600, South Africa.
FEMS Microbiology Letters
|January 16, 2023
Summary
Genetic engineering enables Saccharomyces cerevisiae to produce fumaric acid from malic acid. Modified yeast strains achieved significant fumaric acid yields, showing promise for industrial biorefineries.
Area of Science:
- Biotechnology
- Industrial Microbiology
- Metabolic Engineering
Background:
- Saccharomyces cerevisiae is robust and acid-tolerant, making it suitable for industrial applications.
- Biorefineries can utilize waste streams for high-value organic acid production, like fumaric acid.
- S. cerevisiae does not naturally produce dicarboxylic acids, necessitating genetic modification.
Purpose of the Study:
- To engineer S. cerevisiae for efficient fumaric acid production.
- To investigate factors influencing malic acid to fumaric acid conversion in engineered yeast.
Main Methods:
- Genetic modification of S. cerevisiae, including FUM1 gene disruption.
- Recombinant expression of fumarase and malate transporter genes (MAE1).
- Cultivation of engineered strains under varying conditions (oxygen, secretion signals).
Main Results:
- Engineered strains (ΔFUM1Ckr_fum + mae1 and ΔFUM1(ss)Ckr_fum + mae1) converted malic acid to fumaric acid.
- Yields of 0.98 g/L and 1.11 g/L fumaric acid were achieved under aerobic conditions.
- Fumarase gene source, secretion signals, and oxygen availability impacted conversion efficiency.
Conclusions:
- Genetic engineering can equip S. cerevisiae for fumaric acid biosynthesis.
- Optimized strains show potential for fumaric acid production in industrial settings.
- Further research can enhance yields by refining genetic constructs and process conditions.
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