Carminic acid, an aromatic polyketide compound, is a widely used natural red colorant.Traditionally, carminic acid is extracted from cochineal insects, but the extraction process is labor-intensive and inefficient, leading to insufficient production to meet the growing market demand.In recent years, biosynthetic production of carminic acid has attracted significant attention.In this study, Yarrowia lipolytica was used as a host strain to achieve the de novo synthesis of carminic acid through the heterologous expression and screening of key pathway genes, resulting in a production titer of 0.7 mg/L.Since carminic acid biosynthesis requires two critical precursors, malonyl-CoA and acetyl-CoA, efforts were first made to enhance malonyl-CoA supply by screening ACC1 mutants and introducing a non-native malonyl-CoA biosynthetic pathway (NCM pathway), which increased the carminic acid titer to 1.2 mg/L.Subsequently, acetyl-CoA supply was strengthened by regulating lipid metabolism, further boosting the titer to 1.5 mg/L.Additionally, by improving substrate utilization efficiency and relocating the carminic acid biosynthetic pathway to peroxisomes, the titer reached 1.9 mg/L.Finally, through fed-batch fermentation in a 5 L bioreactor, the production of carminic acid was scaled up to 9.6 mg/L (6.3×10-3mg/g).This study provides a reference for the biosynthesis of carminic acid.
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