通过 UDP-葡萄糖基转移酶催化的级联反应高效生物合成红景天苷。
Highly efficient biosynthesis of salidroside by a UDP-glucosyltransferase-catalyzed cascade reaction.
机构信息
Anhui Engineering Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources, Generic Technology Research Center for Anhui Traditional Chinese Medicine Industry, West Anhui University, Lu'an, 237012, Anhui, China.
Anhui Engineering Research Center for Eco-Agriculture of Traditional Chinese Medicine, Department of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, 237012, Anhui, China.
出版信息
Biotechnol Lett. 2024 Apr;46(2):173-181. doi: 10.1007/s10529-023-03453-0. Epub 2024 Jan 6.
OBJECTIVE
Salidroside is an important plant-derived aromatic compound with diverse biological properties. The main objective of this study was to synthesize salidroside from tyrosol using UDP-glucosyltransferase (UGT) with in situ regeneration of UDP-glucose (UDPG).
RESULTS
The UDP-glucosyltransferase 85A1 (UGT85A1) from Arabidopsis thaliana, which showed high activity and regioselectivity towards tyrosol, was selected for the production of salidroside. Then, an in vitro cascade reaction for in situ regeneration of UDPG was constructed by coupling UGT85A1 to sucrose synthase from Glycine max (GmSuSy). The optimal UGT85A1-GmSuSy activity ratio of 1:2 was determined to balance the efficiency of salidroside production and UDP-glucose regeneration. Different cascade reaction conditions for salidroside production were also determined. Under the optimized condition, salidroside was produced at a titer of 6.0 g/L with a corresponding molar conversion of 99.6% and a specific productivity of 199.1 mg/L/h in a continuous feeding reactor.
CONCLUSION
This is the highest salidroside titer ever reported so far using biocatalytic approach.
目的
红景天苷是一种具有多种生物特性的重要植物衍生芳香化合物。本研究的主要目的是使用 UDP-葡萄糖基转移酶（UGT）与 UDP-葡萄糖（UDPG）原位再生将酪醇合成红景天苷。
结果
选择拟南芥的 UDP-葡萄糖基转移酶 85A1（UGT85A1）用于生产红景天苷，该酶对酪醇表现出高活性和区域选择性。然后，通过将 UGT85A1 与来自 Glycine max（GmSuSy）的蔗糖合酶偶联，构建了用于 UDPG 原位再生的体外级联反应。确定了最佳的 UGT85A1-GmSuSy 活性比为 1:2，以平衡红景天苷生产和 UDP-葡萄糖再生的效率。还确定了不同的级联反应条件用于生产红景天苷。在优化条件下，在连续进料反应器中，红景天苷的产量达到 6.0 g/L，相应的摩尔转化率为 99.6%，比生产率为 199.1 mg/L/h。
结论
这是迄今为止使用生物催化方法报道的最高红景天苷产量。
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