Highly Regioselective and Efficient Biosynthesis of Polydatin by an Engineered UGTBL1-AtSuSy Cascade Reaction
Polydatin, resveratrol-3-O-β-glucoside, possesses various biological activities. However, the regioselective glucosylation of resveratrol by UDP-glycosyltransferases (UGTs) constitutes a persistent problem. In this study, semi-rational design and iterative combinatorial mutagenesis were carried out to screen the mutants of UGTBL1 and the high specificity with the glycosylation of the 3-OH group of resveratrol was explored. The triple mutant I62G/M112D/K143G exhibited near-perfect control of polydatin synthesis (regioselectivity ∼ 99%), and the ratio of polydatin to resveratrol-4'-O-β-glucoside was finally enhanced by 786-fold. Molecular docking revealed that the mutant could form three H-bonds between 3-, 5-, and 4'-OH groups of resveratrol and the residues around the active center, resulting in the oriented-binding of resveratrol. Furthermore, UGTBL1 mutant coupling sucrose synthase AtSuSy can synthesize polydatin at an unprecedented high titer of 10.33 g/L, together with efficient UDPG regeneration (RCmax = 54). This study provides an efficient approach for the regioselective biosynthesis of polydatin.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:69 |
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| Enthalten in: |
Journal of agricultural and food chemistry - 69(2021), 31 vom: 11. Aug., Seite 8695-8702 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Chen, Tianyi [VerfasserIn] |
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| Links: |
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| Themen: |
EC 2.4.- |
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| Anmerkungen: |
Date Completed 12.08.2021 Date Revised 12.08.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1021/acs.jafc.1c02518 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM328644021 |
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| 245 | 1 | 0 | |a Highly Regioselective and Efficient Biosynthesis of Polydatin by an Engineered UGTBL1-AtSuSy Cascade Reaction |
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| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Polydatin, resveratrol-3-O-β-glucoside, possesses various biological activities. However, the regioselective glucosylation of resveratrol by UDP-glycosyltransferases (UGTs) constitutes a persistent problem. In this study, semi-rational design and iterative combinatorial mutagenesis were carried out to screen the mutants of UGTBL1 and the high specificity with the glycosylation of the 3-OH group of resveratrol was explored. The triple mutant I62G/M112D/K143G exhibited near-perfect control of polydatin synthesis (regioselectivity ∼ 99%), and the ratio of polydatin to resveratrol-4'-O-β-glucoside was finally enhanced by 786-fold. Molecular docking revealed that the mutant could form three H-bonds between 3-, 5-, and 4'-OH groups of resveratrol and the residues around the active center, resulting in the oriented-binding of resveratrol. Furthermore, UGTBL1 mutant coupling sucrose synthase AtSuSy can synthesize polydatin at an unprecedented high titer of 10.33 g/L, together with efficient UDPG regeneration (RCmax = 54). This study provides an efficient approach for the regioselective biosynthesis of polydatin | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a UGT-AtSuSy cascade reaction | |
| 650 | 4 | |a glycosyltransferase | |
| 650 | 4 | |a polydatin | |
| 650 | 4 | |a regioselective synthesis | |
| 650 | 4 | |a semi-rational design | |
| 650 | 7 | |a Glucosides |2 NLM | |
| 650 | 7 | |a Stilbenes |2 NLM | |
| 650 | 7 | |a Glycosyltransferases |2 NLM | |
| 650 | 7 | |a EC 2.4.- |2 NLM | |
| 650 | 7 | |a polydatin |2 NLM | |
| 650 | 7 | |a XM261C37CQ |2 NLM | |
| 700 | 1 | |a Chen, Ziyi |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Nan |e verfasserin |4 aut | |
| 700 | 1 | |a Chu, Jianlin |e verfasserin |4 aut | |
| 700 | 1 | |a Fan, Bo |e verfasserin |4 aut | |
| 700 | 1 | |a Cheng, Cheng |e verfasserin |4 aut | |
| 700 | 1 | |a Qin, Song |e verfasserin |4 aut | |
| 700 | 1 | |a He, Bingfang |e verfasserin |4 aut | |
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