Methods and compositions for producing triacetic acid lactone

Genetically modified yeast cells produce malonyl-CoA and triacetic acid lactone in peroxisomes using fatty acids and oils, addressing the cost inefficiency of natural sources and enhancing TAL production for commercial applications.

WO2025259593A1 Publication Date: 2025-12-18PYRONE SYSTEMS INC
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Patent Information

Application Number
PCT/US2025/032863
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2024-06-11
Filing Date
2025-06-09
Publication Date
2025-12-18

AI Technical Summary

Technical Problem

The high cost and inefficiency of producing triacetic acid lactone (TAL) from natural sources like Gerbera daisies limits its commercial application, necessitating a synthetic and bioengineered microbial production method.

Method used

Genetically modified yeast cells are engineered to produce malonyl-coenzyme A and triacetic acid lactone in their peroxisomes using fatty acids, vegetable oils, or alkanes as feedstocks, with enzymes like acyl-CoA oxidase and methylmalonyl-CoA carboxyltransferase localized in the peroxisomes to enhance production.

Benefits of technology

The modified yeast cells efficiently produce malonyl-CoA and TAL, achieving concentrations up to 2,000 pg/L, overcoming the limitations of natural sources and providing a cost-effective biobased substitute for petrochemicals.

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Abstract

Method and compositions for producing malonyl-CoA and / or triacetic acid lactone within peroxisomes using genetically modified yeast cells to metabolize a substrate containing fatty acids, vegetable oil, or alkanes, and genetically modified organisms are provided. Also provided are methods to generate acetyl-CoA, which is retained in the peroxisomes due to genetic modifications that prevent its export therefrom. A modified enzyme with a peroxisomal targeting sequence catalyzes a reaction between acetyl-CoA and oxaloacetate, producing pyruvate and malonyl-CoA. The methods can include deleting carnitine acetyltransferase (CAT2) alleles to prevent acetyl-CoA export from peroxisomes, and using genes encoding methylmalonyl-CoA carboxyltransferase subunits with peroxisomal targeting sequences. Additionally, the method incorporates 2-pyrone synthase genes to further facilitate production processes.
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