Molecular cloning and characterization of a glucosyltransferase catalyzing glucosylation of curcumin in cultured Catharanthus roseus cells.
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 up-regulated 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.
Kaminaga
Yasuhisa
Y
Graduate School of Pharmaceutical Sciences, Nagoya City University, Mizuho-ku, Nagoya 467-8603, Japan.
Sahin
F Pinar
FP
Mizukami
Hajime
H
D002384:Catalysis; D030008:Catharanthus / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003474:Curcumin / Q000031:analogs & derivatives / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation; D007527:Isoenzymes; D010636:Phenols / Q000737:chemistry / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D018076:DNA, Complementary; D029968:Escherichia coli Proteins; D007527:Isoenzymes; D010636:Phenols; D010940:Plant Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D005964:Glucosyltransferases; D003474:Curcumin