Molecular cloning and characterization of a glucosyltransferase catalyzing glucosylation of curcumin in cultured Catharanthus roseus cells

Abstract

Catharanthus roseus cell suspension cultures are capable of converting exogenously supplied curcumin to various glucosides. The glucosylation efficiency is enhanced by addition of methyl jasmonate (MJ) to the cultures prior to curcumin administration. Two cDNAs encoding UDP-glucosyltransferases (CaUGT1 and CaUGT2) were isolated from a cDNA library of cultured C. roseus cells, using a PCR method directed at the conserved UDP-binding domain of plant glycosyltransferases. The sequence identity between their deduced amino acid sequences was 27% %. The expression of both genes was upregulated by addition of MJ to the cell cultures although the mRNA level of CaUGT1 was much lower than that of CaUGT2. The corresponding cDNAs were expressed in Escherichia coli as fusion proteins with maltose-binding protein. The recombinant CaUGT1 exhibited no glucosylation activity with either curcumin or curcumin monoglucoside as substrate, whereas the recombinant CaUGT2 catalyzed the formation of curcumin monoglucoside from curcumin and also conversion of curcumin monoglucoside to curcumin diglucoside. The use of the recombinant CaUGT2 may provide a useful new route for the production of curcumin glucosides.

Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate

“…A number of ginsenosides can be produced by this reaction, and it has been previously reported that there are 35 kinds of ginsenosides in P. ginseng . Unfortunately, the mechanism by which a variety of structures can be produced has yet to be clearly elucidated, although the characterization of genes encoding glucosyltransferase has been documented from several plants (Meesapyodsuk et al 2007;Kaminaga et al 2004;Achnine et al 2005). This may be the result of the innumerable glucosyltransferases encoded in a plant.…”

A single amino acid in the PSPG‐box plays an important role in the catalytic function of CaUGT2 (Curcumin glucosyltransferase), a Group D Family 1 glucosyltransferase from Catharanthus roseus

“…CaUGT2 cDNA was cloned from C. roseus as described previously [23] and UGT73C6 was isolated from Arabidopsis thaliana by RT-PCR, based on the sequence retrieved from the DNA database (Accession no. AAD20155 [At2g36790]).…”

A 7-Deoxyloganetic Acid Glucosyltransferase Contributes a Key Step in Secologanin Biosynthesis in Madagascar Periwinkle

“…PSPGs in the database but not identical to the UGTs previously isolated from periwinkle (Kaminaga et al, 2004;Masada et al, 2009). Using these partial cDNA fragments, we obtained two full-length cDNAs by 59-rapid amplification of cDNA ends (RACE) and designated them as periwinkle UGT6 and UGT7.…”