National Engineering Laboratory for Cereal Fermentation Technology (NELCF), Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
J Agric Food Chem. 2021 Jun 16;69(23):6578-6587. doi: 10.1021/acs.jafc.1c00602. Epub 2021 Jun 1.
Orientin and vitexin are flavone 8-glycosides that exhibit many biological characteristics. This study aimed to establish a two-enzyme-coupled catalytic strategy to enhance the biosynthesis of orientin and vitexin from apigenin and luteolin, respectively. The glucosyltransferase (TcCGT1) gene from was cloned and expressed in BL21(DE3). The optimal activity of TcCGT1 was achieved at pH 9.0 and 37 °C. TcCGT1 was relatively stable over the pH range of 7.0-10.0 at a temperature lower than 45 °C. The coupled catalytic strategy of TcCGT1 and different sucrose synthases was adopted to enhance the production of orientin and vitexin. By optimizing the coupling reaction conditions, orientin and vitexin production successfully achieved 2324.4 and 5524.1 mg/L with a yield of 91.4 and 89.3% (mol/mol), respectively. The coupled catalytic strategy proposed in this study might serve as a promising candidate for the large-scale production of orientin and vitexin in the future.
橙皮苷和牡荆苷是具有多种生物学特性的黄酮 8-糖苷。本研究旨在建立一种双酶偶联催化策略，分别从芹菜素和木樨草素中增强橙皮苷和牡荆苷的生物合成。从 中克隆并在 BL21(DE3)中表达了葡糖基转移酶（TcCGT1）基因。TcCGT1 的最佳活性在 pH9.0 和 37°C 下实现。TcCGT1 在低于 45°C 的温度下在 pH7.0-10.0 的范围内相对稳定。采用 TcCGT1 和不同蔗糖合酶的偶联催化策略来提高橙皮苷和牡荆苷的产量。通过优化偶联反应条件，成功实现了 2324.4 和 5524.1 mg/L 的橙皮苷和牡荆苷产量，产率分别为 91.4%和 89.3%（摩尔/摩尔）。本研究提出的偶联催化策略可能成为未来大规模生产橙皮苷和牡荆苷的有前途的候选方案。
