Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China; School of Biotechnology, Jiangnan University, 1800 Lihu Rd, Wuxi, Jiangsu, 214122, China.
School of Biotechnology, Jiangnan University, 1800 Lihu Rd, Wuxi, Jiangsu, 214122, China.
Metab Eng. 2023 Mar;76:50-62. doi: 10.1016/j.ymben.2023.01.005. Epub 2023 Jan 10.
Carminic acid is a natural red dye extracted from the insect Dactylopius coccus. Due to its ideal dying effect and high safety, it is widely used in food and cosmetics industries. Previous study showed that introduction of polyketide synthase (OKS) from Aloe arborescens, cyclase (ZhuI) and aromatase (ZhuJ) from Streptomyces sp. R1128, and C-glucosyltransferase (UGT2) from D. coccus into Aspergillus nidulans could achieve trace amounts of de novo production. These four genes were introduced into Saccharomyces cerevisiae, but carminic acid was not detected. Analysis of the genome of A. nidulans revealed that 4'-phosphopantetheinyl transferase (NpgA) and monooxygenase (AptC) are essential for de novo biosynthesis of carminic acid in S. cerevisiae. Additionally, endogenous hydroxylase (Cat5) from S. cerevisiae was found to be responsible for hydroxylation of flavokermesic acid to kermesic acid. Therefore, all enzymes and their functions in the biosynthesis of carminic acid were explored and reconstructed in S. cerevisiae. Through systematic pathway engineering, including regulating enzyme expression, enhancing precursor supply, and modifying the β-oxidation pathway, the carminic acid titer in a 5 L bioreactor reached 7580.9 μg/L, the highest yet reported for a microorganism. Heterologous reconstruction of the carminic acid biosynthetic pathway in S. cerevisiae has great potential for de novo biosynthesis of anthraquinone dye.
胭脂虫酸是一种从胭脂虫中提取的天然红色染料。由于其理想的染色效果和高安全性，被广泛应用于食品和化妆品行业。先前的研究表明，将来自龙舌兰的聚酮合酶（OKS）、来自链霉菌的环化酶（ZhuI）和芳香酶（ZhuJ）以及来自胭脂虫的 C-葡萄糖基转移酶（UGT2）引入到aspergillus nidulans 中可以实现痕量的从头生物合成。这四个基因被引入到酿酒酵母中，但没有检测到胭脂虫酸。对aspergillus nidulans 基因组的分析表明，4'-磷酸泛酰巯基乙胺转移酶（NpgA）和单加氧酶（AptC）是酿酒酵母中胭脂虫酸从头生物合成所必需的。此外，还发现酿酒酵母内源性羟化酶（Cat5）负责将 flavokermesic 酸羟化为 kermesic 酸。因此，在酿酒酵母中探索并重建了胭脂虫酸生物合成途径中的所有酶及其功能。通过系统的途径工程，包括调节酶表达、增强前体供应和修饰β-氧化途径，在 5 L 生物反应器中胭脂虫酸的产量达到了 7580.9 μg/L，这是迄今为止微生物报道的最高产量。胭脂虫酸生物合成途径在酿酒酵母中的异源重建为蒽醌染料的从头生物合成提供了巨大的潜力。
