Rebaudioside D (Reb D), a natural high-intensity, low-calorie sweetener, serves as a crucial precursor for synthesizing Rebaudioside M (Reb M), offering significant commercial and health benefits.Research on Reb D is therefore crucial for advancing the development of high-quality sweetener.In this study, a dual-enzyme coupled system was established by constructing a recombinant Escherichia coli strain co-expressing glucosyltransferase EUGT11 and sucrose synthase AtSUS3.This system facilitated the recycling of uridine diphosphate glucose (UDPG), enabling the efficient bioconversion of Reb A to Reb D.Under optimized conditions in a 3 mL glycosylation reaction system containing 100 mmol/L NaH2PO4-Na2HPO4 buffer (pH 7.0), supplementation with 2.4 mg/mL crude enzyme extract, 1.2 g/L Reb A, and 60 g/L sucrose, followed by incubation at 30 ℃ for 20 h, resulted in a maximum Reb D yield of 1 205.81 mg/L, corresponding to an 86.06% conversion rate.This study significantly enhances the synthesis efficiency of Reb D and establishes a robust technical foundation for the cost-effective, large-scale production of high-purity steviol glycosides.
LIU Yuanyuan
,
FENG Mengmeng
,
WEI Tongwei
,
YANG Ting
,
WANG Chengtao
,
ZHAO Liang
,
ZHAO Lei
. Coupled catalysis of glycosyltransferase and sucrose synthase for biosynthesis of Rebaudioside D[J]. Food and Fermentation Industries, 2026
, 52(5)
: 52
-58
.
DOI: 10.13995/j.cnki.11-1802/ts.042858
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