Cascade Catalysis of Glycosyltransferase and Sucrose Synthase to Produce Astragalin

doi:10.19586/j.2095-2341.2022.0212

Current Biotechnology ›› 2023, Vol. 13 ›› Issue (2): 247-256.DOI: 10.19586/j.2095-2341.2022.0212

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Cascade Catalysis of Glycosyltransferase and Sucrose Synthase to Produce Astragalin

Zhekang JIA(), Xinran CHENG, Wenfang DOU()

Life Sciences and Health Engineering College，Jiangnan University，Jiangsu Wuxi 214122，China

Received:2022-12-23 Accepted:2023-03-06 Online:2023-03-25 Published:2023-04-07
Contact: Wenfang DOU

Abstract

Abstract:

Astragalin is a flavonoid 3-O-glycoside compound with various pharmacological activities such as antioxidant， antiviral and antibacterial. In this study， glucosyltransferase from grapefruit and sucrose synthase from soybean were used to convert kaferol to astragalin， while the glycosylation byproduct uridine diphosphate （UDP） was regenerated to UDP-glucose. Through fed-batch operation， the yield of astragalin reached 1 550.1 mg·L^-1， the conversion rate was 86.5%， and the UDP cycle was 34.8 times. After isolation and purification， the results were basically consistent with the literature， and the reaction product was indeed astragalin. The synthesis method of astragalin proposed in this study greatly reduced the reaction cost and provided an new idea for the industrial production of astragalin.

Key words: glycosylation, sucrose synthase, astragalin, kaempferol, UDP-glucose

CLC Number:

Q555

Zhekang JIA, Xinran CHENG, Wenfang DOU. Cascade Catalysis of Glycosyltransferase and Sucrose Synthase to Produce Astragalin[J]. Current Biotechnology, 2023, 13(2): 247-256.

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URL: https://swjsjz.magtechjournal.com/EN/10.19586/j.2095-2341.2022.0212

https://swjsjz.magtechjournal.com/EN/Y2023/V13/I2/247

Figures/Tables 12

Fig. 1 Analysis of purified FGT and GmSUS by SDS-PAGE

Fig. 2 HPLC determination of the glycosyltransferase activity

Fig. 3 Effects of pH, temperature and concentration of DMSO, sucrose, fructose, and UDP on the FGT activity

Fig. 4 HPLC determination of the sucrose synthase activity

Fig. 5 Cascade catalysis

Table 1 Effect of enzyme concentration on the yield of astragaloside

序号	FGT浓度/（mU·mL^-1）	GmSUS浓度/（mU·mL^-1）	紫云英苷产量/（mmol·L^-1）
1	3.68	166	0.063 1
2	18.4	166	0.153 1
3	36.8	166	0.246 7
4	55.2	166	0.235 6
5	73.6	166	0.259 3
6	36.8	830	0.339 9
7	36.8	1 660	0.417 6
8	36.8	2 490	0.456 1
9	36.8	3 320	0.460 2

Fig. 6 Effects of pH, temperature and concentration of DMSO, UDP, sucrose and kaempferol on the cocatalytic reaction

Fig. 7 Production of astragalin by fed-batch

Fig. 8 HPLC analysis of purified product

Fig. 9 LC-MS analysis of purified product

Fig. 10 NMR analysis of purified product

Fig. 11 FGT and GmSUS reacted cascade ally to produce astragalin

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Cascade Catalysis of Glycosyltransferase and Sucrose Synthase to Produce Astragalin

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