Efficient enzymatic production of rebaudioside A from stevioside.
Stevioside and rebaudioside A are the chief diterpene glycosides present in the leaves of Stevia rebaudiana. Rebaudioside A imparts a desirable sweet taste, while stevioside produces a residual bitter aftertaste. Enzymatic synthesis of rebaudioside A from stevioside can increase the ratio of rebaudioside A to stevioside in steviol glycoside products, providing a conceivable strategy to improve the organoleptic properties of steviol glycoside products. Here, we demonstrate the efficient conversion of stevioside to rebaudioside A by coupling the activities of recombinant UDP-glucosyltransferase UGT76G1 from S. rebaudiana and sucrose synthase AtSUS1 from Arabidopsis thaliana. The conversion occurred via regeneration of UDP-glucose by AtSUS1. UDP was applicable as the initial material instead of UDP-glucose for UDP-glucose recycling. The amount of UDP could be greatly reduced in the reaction mixture. Rebaudioside A yield in 30 h with 2.4 mM stevioside, 7.2 mM sucrose, and 0.006 mM UDP was 78%.
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Yu
Y
Wang
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Liangliang
L
Chen
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Yan
Y
Li
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Yangyang
Y
Li
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Ming
M
Yan
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Kequan
K
Chen
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Ning
N
Hao
a College of Biotechnology and Pharmaceutical Engineering , Nanjing Tech University , Nanjing , China.
Lin
L
Xu
D003001:Cloning, Molecular; D045786:Diterpenes, Kaurane / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005524:Food Technology / Q000379:methods; D015870:Gene Expression; D005960:Glucosides / Q000096:biosynthesis* / Q000235:genetics; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D018515:Plant Leaves / Q000737:chemistry / Q000201:enzymology; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D036625:Stevia / Q000737:chemistry* / Q000201:enzymology; D013395:Sucrose / Q000378:metabolism / Q000494:pharmacology; D013549:Sweetening Agents / Q000378:metabolism*; D055697:Taste Perception / Q000502:physiology; D014530:Uridine Diphosphate / Q000378:metabolism / Q000494:pharmacology; D014532:Uridine Diphosphate Glucose / Q000378:metabolism / Q000494:pharmacology
D045786:Diterpenes, Kaurane; D005960:Glucosides; D011994:Recombinant Proteins; D013549:Sweetening Agents; C012043:stevioside; D013395:Sucrose; D014530:Uridine Diphosphate; C025474:rebaudioside A; D005964:Glucosyltransferases; C022699:sucrose synthase; D014532:Uridine Diphosphate Glucose
UDP-glucosyltransferase; UGT76G1; rebaudioside A; stevioside; sucrose synthase