Wu Qi, Chen Ruibing, Zhang Lei
School of Medicine, Shanghai University, 99 Shangda Road, Shanghai, 200444, China.
Department of Pharmaceutical Botany, School of Pharmacy, Naval Medical University, 325 Guohe Rd, Shanghai, 200433, China.
Microb Cell Fact. 2025 Jul 2;24(1):153. doi: 10.1186/s12934-025-02773-2.
Yeast peroxisomes have been engineered as ideal synthetic compartments to enhance the heterologous biosynthesis of natural products, particularly terpenoids and fatty acid derivatives. This advantage is primarily attributed to the rich acetyl-CoA pool generated from the spatially specific fatty acid β-oxidation within peroxisomes. However, their potential for flavonoid biosynthesis has been largely underexplored, primarily due to limited knowledge regarding precursor transport, cofactor availability, and the redox environment in peroxisomes.
In this study, we successfully compartmentalized the biosynthesis of taxifolin, a dihydroflavonol, in Saccharomyces cerevisiae peroxisomes. The result indicated that flavonoid biosynthesis in peroxisome offers a more efficient approach compared to its synthesis in the cytosol. This study managed to expand the application scope of peroxisome compartmentalization to flavonoid biosynthesis. By reinforcing the rate-limiting steps, optimizing cofactor supply and activation of fatty acids, we accomplished the de novo synthesis of taxifolin in peroxisomes for the first time, attaining a titer of 120.3 ± 2.4 mg/L in shake-flask fermentation using a minimal medium.
These findings highlight the feasibility of peroxisomal compartmentalization for flavonoid biosynthesis, providing new insights and a framework for the biosynthesis of other high-value flavonoids using yeast peroxisomes.
酵母过氧化物酶体已被设计成理想的合成区室，以增强天然产物的异源生物合成，特别是萜类化合物和脂肪酸衍生物。这一优势主要归因于过氧化物酶体内空间特异性脂肪酸β-氧化产生的丰富乙酰辅酶A库。然而，它们在黄酮类生物合成方面的潜力在很大程度上尚未得到充分探索，主要是由于对过氧化物酶体中前体运输、辅因子可用性和氧化还原环境的了解有限。
在本研究中，我们成功地将二氢黄酮醇紫杉叶素的生物合成区室化在酿酒酵母过氧化物酶体中。结果表明，与在细胞质中合成相比，过氧化物酶体中的黄酮类生物合成提供了一种更有效的方法。本研究成功地将过氧化物酶体区室化的应用范围扩展到黄酮类生物合成。通过加强限速步骤、优化辅因子供应和脂肪酸活化，我们首次在过氧化物酶体中完成了紫杉叶素的从头合成，在使用基本培养基的摇瓶发酵中达到了120.3±2.4mg/L的滴度。
这些发现突出了过氧化物酶体区室化用于黄酮类生物合成的可行性，为利用酵母过氧化物酶体进行其他高价值黄酮类化合物的生物合成提供了新的见解和框架。
