Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si 31460, Republic of Korea.
Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University, Asan-si 31460, Republic of Korea.
J Microbiol Biotechnol. 2024 May 28;34(5):1154-1163. doi: 10.4014/jmb.2401.01017. Epub 2024 Mar 14.
Glucosylation is a well-known approach to improve the solubility, pharmacological, and biological properties of flavonoids, making flavonoid glucosides a target for large-scale biosynthesis. However, the low yield of products coupled with the requirement of expensive UDP-sugars limits the application of enzymatic systems for large-scale. is a Gram-positive and generally regarded as safe (GRAS) bacteria frequently employed for the large-scale production of amino acids and bio-fuels. Due to the versatility of its cell factory system and its non-endotoxin producing properties, it has become an attractive system for the industrial-scale biosynthesis of alternate products. Here, we explored the cell factory of for efficient glucosylation of flavonoids using apigenin as a model flavonoid, with the heterologous expression of a promiscuous glycosyltransferase, YdhE from and the endogenous overexpression of genes encoding UDP-glucose pyrophosphorylase and encoding phosphoglucomutase involved in the synthesis of UDP-glucose to create a cell factory system capable of efficiently glucosylation apigenin with a high yield of glucosides production. Consequently, the production of various apigenin glucosides was controlled under different temperatures yielding almost 4.2 mM of APG1(apigenin-4'-O-β-glucoside) at 25°C, and 0.6 mM of APG2 (apigenin-7-O-β-glucoside), 1.7 mM of APG3 (apigenin-4',7-O-β-diglucoside) and 2.1 mM of APG4 (apigenin-4',5-O-β-diglucoside) after 40 h of incubation with the supplementation of 5 mM of apigenin and 37°C. The cost-effective developed system could be used to modify a wide range of plant secondary metabolites with increased pharmacokinetic activities on a large scale without the use of expensive UDP-sugars.
糖基化是一种提高黄酮类化合物的水溶性、药理和生物学性质的方法，使黄酮苷成为大规模生物合成的目标。然而，产物的低产量加上对昂贵的 UDP-糖的需求限制了酶系统在大规模生产中的应用。枯草芽孢杆菌是一种革兰氏阳性菌，通常被认为是安全的（GRAS）细菌，常用于大规模生产氨基酸和生物燃料。由于其细胞工厂系统的多功能性及其非内毒素产生特性，它已成为替代产品工业规模生物合成的有吸引力的系统。在这里，我们探索了枯草芽孢杆菌的细胞工厂，以高效地对黄酮类化合物进行糖基化，以芹菜素为模型黄酮类化合物，异源表达一种混杂的糖基转移酶 YdhE 来自，以及内源性过表达参与 UDP-葡萄糖合成的 基因 编码 UDP-葡萄糖焦磷酸化酶和 编码磷酸葡萄糖变位酶，以创建一个 细胞工厂系统，能够有效地以高糖苷产量对芹菜素进行糖基化。因此，在不同温度下控制各种芹菜素糖苷的生产，在 25°C 下产生近 4.2mM 的 APG1（芹菜素-4'-O-β-葡萄糖苷），在 0.6mM 的 APG2（芹菜素-7-O-β-葡萄糖苷）、1.7mM 的 APG3（芹菜素-4'，7-O-β-二葡萄糖苷）和 2.1mM 的 APG4（芹菜素-4'，5-O-β-二葡萄糖苷），在补充 5mM 的芹菜素和 37°C 孵育 40 小时后。开发的具有成本效益的系统可用于修饰广泛的植物次生代谢物，提高药代动力学活性，而无需使用昂贵的 UDP-糖。
