Method for enzymatically synthesizing astragalin

Abstract

The invention provides a method for enzymatically synthesizing astragalin. The method comprises the following steps that 1, key enzymes needed in the synthesizing process of astragalin are cloned, expressed and purified, wherein the key enzymes comprise glycogen phosphorylase GP, glucose pyrophosphorylase GalU, flavanone-3-hydroxylase F3H, flavanone synthase FLS1 and flavonoid3-O-glucanotransferase UGT78K2; 2, glycogen Gn is synthesized into glucose-1-phosphoric acid G-1-P under the GP effect; 3, G-1-P is synthesized into uridine diphosphate UDPG under the GalU effect; 4, naringenin NRN is synthesized into dihydro kaempferol DHK under the F3H effect; 5, DHK is synthesized into kaempferol KMF under the FLS1 effect; 6, KMF and UDPG are synthesized into astragalin under the UGT78K2 effect. According to the method, few steps are adopted, the operation condition is mild, few side products are generated, the yield is large, pollution is avoided, and the production cost is remarkably reduced.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and utilizes an enzymatic method to synthesize a target compound, ascarin, with a classification number of C12P. Background technique [0002] Shivet glycoside is a natural flavonoid compound widely present in medicinal plants. It has anti-inflammatory, anti-oxidation, cardiotonic, analgesic, antibacterial, anti-allergic, anti-hepatotoxic effects, and can enhance the body's resistance, Stimulate the production of interferon, anti-arrhythmia, dilate blood vessels, protect the myocardium and so on. Although asiaticoside has great potential to treat various human diseases, the way to obtain asiacoside is relatively single at present. [0003] One of the commonly used acquisition methods at home and abroad is the organic solvent extraction method. This method has cumbersome and complicated steps, a long cycle, low yield, high cost, and requires a large amount of organic solvents in the separatio...

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Problems solved by technology

[0004] Another commonly used method to obtain astragalus is the chemical synthesis method. The reaction conditions of this method are harsh, the synthesis process is complicated, the reaction catalysts are mostly poisonous substances, and the regioselectivity is poor, a large number of by-products are easily produced, and the yield is low. The group protection and glycosylation modification used in the reaction process increase the reaction steps, prolong the reaction time, and increase the number of reagents introduced, especially the conditions of glycosylation are extremely harsh and the specificity is not high, which not only increases The difficulty of production increases the cost of production, and these unfavorable factors greatly limit the industrial production of ascarin

For example, Toshihiro Kumazawa et al. used polyhydroxyacetophenone as raw material and boron trifluoride-diethyl ether as a catalyst to raise the reaction temperature from -78°C to -30°C for glycosidation, and other monoglycosides and polyglycosides were easily formed during the synthesis process. Compounds increase the by-products and decrease the yield

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