Engineering regioselectivity of glycosyltransferase for efficient polydatin synthesis.
Food Chem
; 460(Pt 2): 140698, 2024 Dec 01.
Article
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| MEDLINE
| ID: mdl-39098192
ABSTRACT
Resveratrol is a promising functional ingredient applied in food products. However, low bioavailability and poor water solubility, which can be improved by glycosylation, hinder its application. A uridine diphosphate-dependent glycosyltransferase (UGT) from Bacillus subtilis 168 (named UGTBS) presents potential application for resveratrol glycosylation; nonetheless, imprecise regioselectivity renders the synthesis of resveratrol-3-O-ß-D-glucoside (polydatin) difficult. Therefore, molecular evolution was applied to UGTBS. A triple mutant Y14I/I62G/M315W was developed for 3-OH glycosylation of resveratrol and polydatin accounted for 91% of the total product. Kinetic determination and molecular docking indicated that the enhancement of hydrogen bond interaction and altered conformation of the binding pocket increases the enzyme's affinity for the 3-OH group, stabilizing the enzyme-substrate intermediate and promoting polydatin formation. Furthermore, a fed-batch cascade reaction by periodic addition of resveratrol was conducted and nearly 20 mM polydatin was obtained. The mutant Y14I/I62G/M315W can be used for polydatin manufacture.
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MEDLINE
Asunto principal:
Estilbenos
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Bacillus subtilis
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Glicosiltransferasas
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Simulación del Acoplamiento Molecular
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Glucósidos
Idioma:
En
Revista:
Food Chem
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Food chem
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Food chemistry
Año:
2024
Tipo del documento:
Article