The Activity of ArabidopsisGlycosyltransferases toward Salicylic Acid, 4-Hydroxybenzoic Acid, and Other Benzoates

Abstract

Benzoates are a class of natural products containing compounds of industrial and strategic importance. In plants, the compounds exist in free form and as conjugates to a wide range of other metabolites such as glucose, which can be attached to the carboxyl group or to specific hydroxyl groups on the benzene ring. These glucosylation reactions have been studied for many years, but to date only one gene encoding a benzoate glucosyltransferase has been cloned. A phylogenetic analysis of sequences in theArabidopsis genome revealed a large multigene family of putative glycosyltransferases containing a consensus sequence typically found in enzymes transferring glucose to small molecular weight compounds such as secondary metabolites. Ninety of these sequences have now been expressed as recombinant proteins in Escherichia coli, and their in vitro catalytic activities toward benzoates have been analyzed. The data show that only 14 proteins display activity toward 2-hydroxybenzoic acid, 4-hydroxybenzoic acid, and 3,4-dihydroxybenzoic acid. Of these, only two enzymes are active toward 2-hydroxybenzoic acid, suggesting they are theArabidopsis salicylic acid glucosyltransferases. All of the enzymes forming glucose esters with the metabolites were located in Group L of the phylogenetic tree, whereas those formingO-glucosides were dispersed among five different groups. Catalytic activities were observed toward glucosylation of the 2-, 3-, or 4-hydroxyl group on the ring. To further explore their regioselectivity, the 14 enzymes were analyzed against benzoic acid, 3-hydroxybenzoic acid, 2,3-, 2,4-, 2,5-, and 2,6-dihydroxybenzoic acid. The data showed that glycosylation of specific sites could be positively or negatively influenced by the presence of additional hydroxyl groups on the ring. This study provides new tools for biotransformation reactions in vitro and a basis for engineering benzoate metabolism in plants. Benzoates are a class of natural products containing compounds of industrial and strategic importance. In plants, the compounds exist in free form and as conjugates to a wide range of other metabolites such as glucose, which can be attached to the carboxyl group or to specific hydroxyl groups on the benzene ring. These glucosylation reactions have been studied for many years, but to date only one gene encoding a benzoate glucosyltransferase has been cloned. A phylogenetic analysis of sequences in theArabidopsis genome revealed a large multigene family of putative glycosyltransferases containing a consensus sequence typically found in enzymes transferring glucose to small molecular weight compounds such as secondary metabolites. Ninety of these sequences have now been expressed as recombinant proteins in Escherichia coli, and their in vitro catalytic activities toward benzoates have been analyzed. The data show that only 14 proteins display activity toward 2-hydroxybenzoic acid, 4-hydroxybenzoic acid, and 3,4-dihydroxybenzoic acid. Of these, only two enzymes are active toward 2-hydroxybenzoic acid, suggesting they are theArabidopsis salicylic acid glucosyltransferases. All of the enzymes forming glucose esters with the metabolites were located in Group L of the phylogenetic tree, whereas those formingO-glucosides were dispersed among five different groups. Catalytic activities were observed toward glucosylation of the 2-, 3-, or 4-hydroxyl group on the ring. To further explore their regioselectivity, the 14 enzymes were analyzed against benzoic acid, 3-hydroxybenzoic acid, 2,3-, 2,4-, 2,5-, and 2,6-dihydroxybenzoic acid. The data showed that glycosylation of specific sites could be positively or negatively influenced by the presence of additional hydroxyl groups on the ring. This study provides new tools for biotransformation reactions in vitro and a basis for engineering benzoate metabolism in plants. UDP-glycosyltransferase high pressure liquid chromatography benzoic acid hydroxybenzoic acid dihydroxybenzoic acid indole-3-acetic acid Glycosyltransferases catalyze the transfer of sugar from nucleoside diphosphate donors to a wide range of acceptor molecules (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar). In recent years, the enzymes have been classified in a number of different ways including overall sequence homology (2Campbell J.A. Davies G.J. Bulone V. Henrissat B. Biochem. J. 1997; 326: 929-942Crossref PubMed Scopus (624) Google Scholar) and the presence of a variety of common motifs (3Bairoch A. Nucleic Acids Res. 1991; 19: 2241-2245Crossref PubMed Scopus (518) Google Scholar, 4Hughes J. Hughes M.A. DNA Seq. 1994; 5: 41-49Crossref PubMed Scopus (164) Google Scholar, 5Mackenzie P.I. Owen I.S. Burchell B. Bock K.W. Bairoch A. Bélanger A. Fournel-Gigleux S. Green M. Hum D.W. Iyanagi T. Lancet D. Louisot P. Magdalou J. Chowdhury J.R. Ritter J.K. Schachter H. Tephly T.R. Tipton K.F. Nebert D.W. Pharmacogenetics. 1997; 7: 255-269Crossref PubMed Scopus (992) Google Scholar, 6Busch C. Hofmann F. Selzer J. Munro S. Jeckel D. Aktories K. J. Biol. Chem. 1998; 273: 19566-19572Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar, 7Henrissat B. Davies G.J. Plant Physiol. 2000; 124: 1515-1519Crossref PubMed Scopus (226) Google Scholar). Analysis of sequence homology has led to the classification of 54 families with a ubiquitous distribution throughout living organisms (7Henrissat B. Davies G.J. Plant Physiol. 2000; 124: 1515-1519Crossref PubMed Scopus (226) Google Scholar). One of the largest families is Family 1, in which many of its members contain a consensus sequence found toward the C termini of the proteins thought to be related to nucleoside diphosphate-sugar binding (8Ross J. Biol. Google Scholar). In a study of the consensus has been to a large multigene family of P.I. Owen I.S. Burchell B. Bock K.W. Bairoch A. Bélanger A. Fournel-Gigleux S. Green M. Hum D.W. Iyanagi T. Lancet D. Louisot P. Magdalou J. Chowdhury J.R. Ritter J.K. Schachter H. Tephly T.R. Tipton K.F. Nebert D.W. Pharmacogenetics. 1997; 7: 255-269Crossref PubMed Scopus (992) Google Scholar). In many enzymes are that contain consensus but in to the the enzymes as the in the transfer (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar, P. T. PubMed Scopus Google containing the consensus sequence in a variety of including of secondary and of such as and P. T. PubMed Scopus Google Scholar, K. J. Plant Scopus Google Scholar, A. H. Biochem. 1994; PubMed Scopus Google Scholar, Plant 1991; Scopus Google Scholar). of a sugar the acceptor can to in the activity of the acceptor as as in its glucosylation of such as acid, and their activity K. J. Plant Scopus Google Scholar). the acceptor can the and a by such as those in the of the which is to be the for of the in J. Google have the consensus sequence to the genome data of to the number of putative enzymes of class in a S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of a large multigene family of sequences (8Ross J. Biol. Google Scholar). one of of their have expressed the sequences in Escherichia and recombinant proteins for catalytic activity in This has led to the of encoding enzymes that the of A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) and two indole-3-acetic acid and acid M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The have a for the of the their and their of the gene encoding forming the glucose of and a of as a by the the to a M. J. and D. J. for M. J. and D. J. for study now the analysis of to a new class of the benzoates is in these from the of their in the such as the of salicylic acid in in Plant Biol. 2000; PubMed Scopus Google and from their in such as the of 4-hydroxybenzoic acid as a and 3,4-dihydroxybenzoic acid as D. S. B. J. Scopus Google Scholar, S. Scopus Google Scholar, K. H. Biol. PubMed Scopus Google Scholar, B. H. S. K. J. Biol. PubMed Scopus Google Scholar, Biol. PubMed Scopus Google Scholar). Benzoates to explore the of glucosylation that can on the benzene and a wide variety of compounds are for as The of hydroxyl groups is to many of the such as their and their to free Biol. PubMed Scopus Google Scholar, Res. Google Scholar, H. J. P. M. J. Chem. 1991; Scopus Google the carboxyl group of the a glucose whereas glucosylation one of the hydroxyl on the the compounds are in which the and hydroxyl groups are Biochem. Biol. 1998; Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, M. K. K. K. K. Scopus Google Scholar, A. M. T. Scopus Google Scholar, S. K. Scopus Google Scholar, 7: Scopus Google Scholar). esters of are to be in which the high of the glucose transfer of the a further acceptor D. Scopus Google Scholar). in metabolism is that of glucose is in the of and V. D. PubMed Scopus Google Scholar, D. W. B. Scholar, D. PubMed Scopus Google Scholar, D. Google Scholar). in the glucose of is in the of Biochem. J. PubMed Scopus Google Scholar). In are thought to be of the and are those that are from the or throughout the (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar). In of the of the to high of in the that is by glucosylation and the A. Biochem. 1991; PubMed Scopus Google Scholar, H. M. Scopus Google Scholar, K. H. M. Scopus Google Scholar, K. K. M. M. Scopus Google study have the activity of recombinant toward a of benzoates for their and of their hydroxyl groups on the ring. The 14 that activities toward these compounds have been further analyzed against additional to the of their catalytic such as those in glycosylation typically transfer glucose from to small acceptor is in class of enzymes they a of glycosylation that is to by The enzymes a to of secondary metabolism in and to the of to industrial these glycosyltransferases have been from and have been in J. Biol. Google and P. T. PubMed Scopus Google Scholar). the to has been of the and of its This can many of the enzymes are in in the these the to salicylic acid glucosyltransferase J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google V. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and T. D. Plant J. 19: PubMed Scopus Google Scholar) are of the has led to is to of to and data to sequences that to those encoding This is with the of a of many In analysis of the genome revealed sequences containing a consensus that they to glycosyltransferases (8Ross J. Biol. Google Scholar, S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) such as those that secondary and a of these sequences to enzymes and their recombinant proteins in were for catalytic activities in This led to the of encoding of the including and A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The have that in multigene families can be and the of of sequence the glucosyltransferase to of the sequences with including that with the sequence homology to the sequence M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google the study have the of in vitro for catalytic activities toward class of natural the This class many compounds of for their activities in the and their as salicylic acid, is in and is to and a wide of J. Plant Biol. PubMed Scopus Google Scholar). of salicylic acid in has been the of the and glucose Biochem. Biol. 1998; Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been that glucosylation of salicylic acid. has to its as a D. S. B. J. Scopus Google Scholar). The is the in of a natural of D. Chem. PubMed Scopus Google Scholar). is a its is located in the K. K. M. M. Scopus Google Scholar). This in is to be of the is a for the of from the is benzoate of its activity K. H. Biol. PubMed Scopus Google Scholar). is to that of have in the T. T. H. S. J. Scopus Google Scholar, 1997; PubMed Scopus Google Scholar). The glycosyltransferases in the of these compounds have been studied many In the of the has been to from and to its The recombinant vitro and to form the glucose in to the J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been from but as the gene has been 1997; Scopus Google Scholar). on of is the has been in but the has been the benzoates in a of recombinant proteins from the multigene have now 14 encoding enzymes with catalytic activity toward one or of these The study has a by the glycosyltransferases that is in only two of the enzymes can the of the benzene and one of the can the carboxyl group of salicylic acid, is specific the of salicylic acid is to the phylogenetic the of the that is is that containing and This of from sequence homology with that for the glucosyltransferase M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of the catalytic activity of the and toward salicylic acid, is that is the of the to the of and for the many enzymes were found to be for the or the In the as as the of glucose the in vitro activity of the enzymes found to be influenced by the of additional hydroxyl groups to the glucosylation The could be or for were study the of a to glycosyltransferases that are active toward of importance. a is for the enzymes as or further their catalytic activities by of their in biotransformation is by for high of the J. Res. PubMed Scopus Google Scholar). a of catalytic activity in vitro provides a new for the of the enzymes in the In of the glucosyltransferase to In a now be to the of salicylic acid in and to study in the of of enzymes and the such as in the and of M. S. K. Plant Physiol. PubMed Scopus Google Scholar, S. Plant Physiol. 1998; PubMed Scopus Google Scholar, S. A. K. K. A. Plant Biol. PubMed Scopus Google Scholar) a of engineering the of of the and its in the Glycosyltransferases catalyze the transfer of sugar from nucleoside diphosphate donors to a wide range of acceptor molecules (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar). In recent years, the enzymes have been classified in a number of different ways including overall sequence homology (2Campbell J.A. Davies G.J. Bulone V. Henrissat B. Biochem. J. 1997; 326: 929-942Crossref PubMed Scopus (624) Google Scholar) and the presence of a variety of common motifs (3Bairoch A. Nucleic Acids Res. 1991; 19: 2241-2245Crossref PubMed Scopus (518) Google Scholar, 4Hughes J. Hughes M.A. DNA Seq. 1994; 5: 41-49Crossref PubMed Scopus (164) Google Scholar, 5Mackenzie P.I. Owen I.S. Burchell B. Bock K.W. Bairoch A. Bélanger A. Fournel-Gigleux S. Green M. Hum D.W. Iyanagi T. Lancet D. Louisot P. Magdalou J. Chowdhury J.R. Ritter J.K. Schachter H. Tephly T.R. Tipton K.F. Nebert D.W. Pharmacogenetics. 1997; 7: 255-269Crossref PubMed Scopus (992) Google Scholar, 6Busch C. Hofmann F. Selzer J. Munro S. Jeckel D. Aktories K. J. Biol. Chem. 1998; 273: 19566-19572Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar, 7Henrissat B. Davies G.J. Plant Physiol. 2000; 124: 1515-1519Crossref PubMed Scopus (226) Google Scholar). Analysis of sequence homology has led to the classification of 54 families with a ubiquitous distribution throughout living organisms (7Henrissat B. Davies G.J. Plant Physiol. 2000; 124: 1515-1519Crossref PubMed Scopus (226) Google Scholar). One of the largest families is Family 1, in which many of its members contain a consensus sequence found toward the C termini of the proteins thought to be related to nucleoside diphosphate-sugar binding (8Ross J. Biol. Google Scholar). In a study of the consensus has been to a large multigene family of P.I. Owen I.S. Burchell B. Bock K.W. Bairoch A. Bélanger A. Fournel-Gigleux S. Green M. Hum D.W. Iyanagi T. Lancet D. Louisot P. Magdalou J. Chowdhury J.R. Ritter J.K. Schachter H. Tephly T.R. Tipton K.F. Nebert D.W. Pharmacogenetics. 1997; 7: 255-269Crossref PubMed Scopus (992) Google Scholar). In many enzymes are that contain consensus but in to the the enzymes as the in the transfer (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar, P. T. PubMed Scopus Google Scholar). The containing the consensus sequence in a variety of including of secondary and of such as and P. T. PubMed Scopus Google Scholar, K. J. Plant Scopus Google Scholar, A. H. Biochem. 1994; PubMed Scopus Google Scholar, Plant 1991; Scopus Google Scholar). of a sugar the acceptor can to in the activity of the acceptor as as in its glucosylation of such as acid, and their activity K. J. Plant Scopus Google Scholar). the acceptor can the and a by such as those in the of the which is to be the for of the in J. Google Scholar). have the consensus sequence to the genome data of to the number of putative enzymes of class in a S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of a large multigene family of sequences (8Ross J. Biol. Google Scholar). one of of their have expressed the sequences in Escherichia and recombinant proteins for catalytic activity in This has led to the of encoding enzymes that the of A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) and two indole-3-acetic acid and acid M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The have a for the of the their and their of the gene encoding forming the glucose of and a of as a by the the to a M. J. and D. J. for M. J. and D. J. for This study now the analysis of to a new class of the benzoates is in these from the of their in the such as the of salicylic acid in in Plant Biol. 2000; PubMed Scopus Google and from their in such as the of 4-hydroxybenzoic acid as a and 3,4-dihydroxybenzoic acid as D. S. B. J. Scopus Google Scholar, S. Scopus Google Scholar, K. H. Biol. PubMed Scopus Google Scholar, B. H. S. K. J. Biol. PubMed Scopus Google Scholar, Biol. PubMed Scopus Google Scholar). Benzoates to explore the of glucosylation that can on the benzene and a wide variety of compounds are for as The of hydroxyl groups is to many of the such as their and their to free Biol. PubMed Scopus Google Scholar, Res. Google Scholar, H. J. P. M. J. Chem. 1991; Scopus Google Scholar). the carboxyl group of the a glucose whereas glucosylation one of the hydroxyl on the the compounds are in which the and hydroxyl groups are Biochem. Biol. 1998; Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, M. K. K. K. K. Scopus Google Scholar, A. M. T. Scopus Google Scholar, S. K. Scopus Google Scholar, 7: Scopus Google Scholar). esters of are to be in which the high of the glucose transfer of the a further acceptor D. Scopus Google Scholar). in metabolism is that of glucose is in the of and V. D. PubMed Scopus Google Scholar, D. W. B. Scholar, D. PubMed Scopus Google Scholar, D. Google Scholar). in the glucose of is in the of Biochem. J. PubMed Scopus Google Scholar). In are thought to be of the and are those that are from the or throughout the (1Hostel W. The Biochemistry of Plants. 7. Academic Press, Inc., New York1981: 725-753Google Scholar). In of the of the to high of in the that is by glucosylation and the A. Biochem. 1991; PubMed Scopus Google Scholar, H. M. Scopus Google Scholar, K. H. M. Scopus Google Scholar, K. K. M. M. Scopus Google Scholar). In study have the activity of recombinant toward a of benzoates for their and of their hydroxyl groups on the ring. The 14 that activities toward these compounds have been further analyzed against additional to the of their catalytic such as those in glycosylation typically transfer glucose from to small acceptor is in class of enzymes they a of glycosylation that is to by The enzymes a to of secondary metabolism in and to the of to industrial these glycosyltransferases have been from and have been in J. Biol. Google and P. T. PubMed Scopus Google Scholar). the to has been of the and of its This can many of the enzymes are in in the these the to salicylic acid glucosyltransferase J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google V. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and T. D. Plant J. 19: PubMed Scopus Google Scholar) are of the has led to is to of to and data to sequences that to those encoding This is with the of a of many In analysis of the genome revealed sequences containing a consensus that they to glycosyltransferases (8Ross J. Biol. Google Scholar, S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) such as those that secondary and a of these sequences to enzymes and their recombinant proteins in were for catalytic activities in This led to the of encoding of the including and A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The have that in multigene families can be and the of of sequence the glucosyltransferase to of the sequences with including that with the sequence homology to the sequence M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google the study have the of in vitro for catalytic activities toward class of natural the This class many compounds of for their activities in the and their as salicylic acid, is in and is to and a wide of J. Plant Biol. PubMed Scopus Google Scholar). of salicylic acid in has been the of the and glucose Biochem. Biol. 1998; Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been that glucosylation of salicylic acid. has to its as a D. S. B. J. Scopus Google Scholar). The is the in of a natural of D. Chem. PubMed Scopus Google Scholar). is a its is located in the K. K. M. M. Scopus Google Scholar). This in is to be of the is a for the of from the is benzoate of its activity K. H. Biol. PubMed Scopus Google Scholar). is to that of have in the T. T. H. S. J. Scopus Google Scholar, 1997; PubMed Scopus Google Scholar). The glycosyltransferases in the of these compounds have been studied many In the of the has been to from and to its The recombinant vitro and to form the glucose in to the J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been from but as the gene has been 1997; Scopus Google Scholar). on of is the has been in but the has been the benzoates in a of recombinant proteins from the multigene have now 14 encoding enzymes with catalytic activity toward one or of these The study has a by the glycosyltransferases that is in only two of the enzymes can the of the benzene and one of the can the carboxyl group of salicylic acid, is specific the of salicylic acid is to the phylogenetic the of the that is is that containing and This of from sequence homology with that for the glucosyltransferase M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of the catalytic activity of the and toward salicylic acid, is that is the of the to the of and for the many enzymes were found to be for the or the In the as as the of glucose the in vitro activity of the enzymes found to be influenced by the of additional hydroxyl groups to the glucosylation The could be or for were study the of a to glycosyltransferases that are active toward of importance. a is for the enzymes as or further their catalytic activities by of their in biotransformation is by for high of the J. Res. PubMed Scopus Google Scholar). a of catalytic activity in vitro provides a new for the of the enzymes in the In of the glucosyltransferase to In a now be to the of salicylic acid in and to study in the of of enzymes and the such as in the and of M. S. K. Plant Physiol. PubMed Scopus Google Scholar, S. Plant Physiol. 1998; PubMed Scopus Google Scholar, S. A. K. K. A. Plant Biol. PubMed Scopus Google Scholar) a of engineering the of of the and its in the Plant such as those in glycosylation typically transfer glucose from to small acceptor is in class of enzymes they a of glycosylation that is to by The enzymes a to of secondary metabolism in and to the of to industrial these glycosyltransferases have been from and have been in J. Biol. Google and P. T. PubMed Scopus Google Scholar). the to has been of the and of its This can many of the enzymes are in in the these the to salicylic acid glucosyltransferase J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google V. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and T. D. Plant J. 19: PubMed Scopus Google Scholar) are of the has led to is to of to and data to sequences that to those encoding This is with the of a of many In analysis of the genome revealed sequences containing a consensus that they to glycosyltransferases (8Ross J. Biol. Google Scholar, S. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) such as those that secondary and a of these sequences to enzymes and their recombinant proteins in were for catalytic activities in This led to the of encoding of the including and A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). The have that in multigene families can be and the of of sequence the glucosyltransferase to of the sequences with including that with the sequence homology to the sequence M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In the study have the of in vitro for catalytic activities toward class of natural the This class many compounds of for their activities in the and their as salicylic acid, is in and is to and a wide of J. Plant Biol. PubMed Scopus Google Scholar). of salicylic acid in has been the of the and glucose Biochem. Biol. 1998; Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been that glucosylation of salicylic acid. has to its as a D. S. B. J. Scopus Google Scholar). The is the in of a natural of D. Chem. PubMed Scopus Google Scholar). is a its is located in the K. K. M. M. Scopus Google Scholar). This in is to be of the is a for the of from the is benzoate of its activity K. H. Biol. PubMed Scopus Google Scholar). is to that of have in the T. T. H. S. J. Scopus Google Scholar, 1997; PubMed Scopus Google Scholar). The glycosyltransferases in the of these compounds have been studied many In the of the has been to from and to its The recombinant vitro and to form the glucose in to the J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). has been from but as the gene has been 1997; Scopus Google Scholar). on of is the has been in but the has been the benzoates in a of recombinant proteins from the multigene have now 14 encoding enzymes with catalytic activity toward one or of these The study has a by the glycosyltransferases that is in only two of the enzymes can the of the benzene and one of the can the carboxyl group of salicylic acid, is specific the of salicylic acid is to the phylogenetic the of the that is is that containing and This of from sequence homology with that for the glucosyltransferase M. J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of the catalytic activity of the and toward salicylic acid, is that is the of the In to the of and for the many enzymes were found to be for the or the In the as as the of glucose the in vitro activity of the enzymes found to be influenced by the of additional hydroxyl groups to the glucosylation The could be or for were This study the of a to glycosyltransferases that are active toward of importance. a is for the enzymes as or further their catalytic activities by of their in biotransformation is by for high of the J. Res. PubMed Scopus Google Scholar). a of catalytic activity in vitro provides a new for the of the enzymes in the In of the glucosyltransferase to In a now be to the of salicylic acid in and to study in the of of enzymes and the such as in the and of M. S. K. Plant Physiol. PubMed Scopus Google Scholar, S. Plant Physiol. 1998; PubMed Scopus Google Scholar, S. A. K. K. A. Plant Biol. PubMed Scopus Google Scholar) a of engineering the of of the and its in the P. and S. of for glucose conjugates of and salicylic acid, and and for