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Four glucosyltransferases from rice: cDNA cloning, expression, and characterization
Jae Hyung Ko1, Bong Gyu Kim, Jeong Ho Kim
1Division of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul, Korea.
Journal of Plant Physiology
|March 17, 2007
Summary
Four rice UDP-dependent glucosyltransferase (UGT) enzymes were studied for their ability to glycosylate flavonoids. UGT706C1, UGT706D1, and UGT707A3 likely synthesize kaempferol and quercetin glucosides in vivo.
Area of Science:
- Plant Biochemistry
- Molecular Biology
- Enzymology
Background:
- UDP-dependent glucosyltransferases (UGTs) play crucial roles in plant secondary metabolism.
- Flavonoids are important plant compounds with diverse biological activities.
- Understanding UGT function is key to elucidating plant metabolic pathways.
Purpose of the Study:
- To characterize four rice UGT genes (UGT706C1, UGT706D1, UGT707A3, UGT709A4) regarding their substrate specificity towards various flavonoids.
- To determine the potential in vivo function of these UGTs in rice flavonoid glycosylation.
Main Methods:
- Cloning and expression of four rice UGT genes in Escherichia coli.
- Purification of recombinant UGT enzymes.
- In vitro enzymatic assays using various flavonoids (apigenin, daidzein, genistein, kaempferol, luteolin, naringenin, quercetin) as potential glucose acceptors.
- Analysis of glycosylation products using HPLC, UV, and NMR spectroscopy.
Main Results:
- UGT706C1 and UGT707A3 specifically glycosylated kaempferol and quercetin at the 3-hydroxyl group.
- UGT709A4 exhibited specificity for isoflavonoids (genistein, daidzein), glycosylating the 7-hydroxyl group.
- UGT706D1 demonstrated broad substrate specificity, glycosylating various flavonoid classes at different positions.
Conclusions:
- The characterized rice UGTs exhibit distinct substrate preferences, indicating specialized roles in flavonoid metabolism.
- UGT706C1, UGT706D1, and UGT707A3 are likely involved in the biosynthesis of kaempferol and quercetin glucosides in rice.
- These findings contribute to understanding the complex network of flavonoid glycosylation in plants.
