Identification, Properties and Genetic Control of p-CoumaroylCoenzyme A, 3-Hydroxylase Isolated from Petals of Silene dioica

Kamsteeg, John; Brederode, J. van; Verschuren, Paul M.; Nigtevecht, G. van

doi:10.1016/s0044-328x(81)80178-x

Cited by 43 publications

(16 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was obvious that no P450 catalyzed this reaction since incubation of microsomes from various plants with radiolabeled cinnamic acid led to p-coumaric acid, but caffeic acid was never produced, even at low levels. Studies of the 3-hydroxylation reaction by several laboratories then led to evidence that parallel pathways may exist, acting at the level of conjugated hydroxycinnamic acids such as esters of CoA (46,47), shikimate and quinate (7,9), phenyl lactate (48), and glucose (49). The CoA ester of p-coumaric acid was recently considered as the best potential substrate since the methylation of caffeate to ferulate, which is the next step in the pathway, was shown to occur mainly on the ester of CoA (17, 24 -26, 28, 43).…”

Section: Discussionmentioning

confidence: 99%

“…Three enzymes were so far described to catalyze the 3-hydroxylation of coumaroyl-CoA. One is a nonspecific polyphenol oxidase (50), the second a soluble FAD-dependent hydroxylase (46), and the third a Zn 2ϩ -dependent dioxygenase that was described to be inactive at a normal cytoplasmic pH (47). None of them was characterized at the molecular level or was regarded as a top candidate for catalyzing the reaction in planta.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

CYP98A3 from Arabidopsis thaliana Is a 3′-Hydroxylase of Phenolic Esters, a Missing Link in the Phenylpropanoid Pathway

Schoch

Goepfert

Morant

et al. 2001

Journal of Biological Chemistry

405

370

View full text Add to dashboard Cite

The 4-and 5-hydroxylations of phenolic compounds in plants are catalyzed by cytochrome P450 enzymes. The 3-hydroxylation step leading to the formation of caffeic acid from p-coumaric acid remained elusive, however, alternatively described as a phenol oxidase, a dioxygenase, or a P450 enzyme, with no decisive evidence for the involvement of any in the reaction in planta. In this study, we show that the gene encoding CYP98A3, which was the best possible P450 candidate for a 3-hydroxylase in the Arabidopsis genome, is highly expressed in inflorescence stems and wounded tissues. Recombinant CYP98A3 expressed in yeast did not metabolize free pcoumaric acid or its glucose or CoA esters, p-coumaraldehyde, or p-coumaryl alcohol, but very actively converted the 5-O-shikimate and 5-O-D-quinate esters of trans-p-coumaric acid into the corresponding caffeic acid conjugates. The shikimate ester was converted four times faster than the quinate derivative. Antibodies directed against recombinant CYP98A3 specifically revealed differentiating vascular tissues in stem and root. Taken together, these data show that CYP98A3 catalyzes the synthesis of chlorogenic acid and very likely also the 3-hydroxylation of lignin monomers. This hydroxylation occurs on depsides, the function of which was so far not understood, revealing an additional and unexpected level of networking in lignin biosynthesis.Systematic genome sequencing is revealing a large number of orphan genes and their phylogenetic relatedness to genes with characterized function. EST 1 sequences, on the other hand, are providing preliminary information on levels, patterns of expression, and conservation of genes among species. Taken together, such information can be exploited as a clue to gene function and to track down missing steps in important pathways.The sequencing of the whole genome of Arabidopsis thaliana has revealed 273 cytochrome P450 genes distributed into 45 families and subfamilies (drnelson.utmem.edu/CytochromeP450. html, www.biobase.dk/P450/). P450 proteins thus form the largest superfamily of enzymes involved in plant metabolism, but the function of 80% of these enzymes is still unknown. Our attention was first drawn to the CYP98 family by its phylogeny and structure. An analysis of P450 phylogeny in A. thaliana (Fig. 1) shows that the CYP98 family is most closely related to CYP73A5, coding for the cinnamic-acid 4-hydroxylase, the second enzyme and first P450 in the phenylpropanoid pathway (1). CYP73A5 and the CYP98 family seem to have evolved from the same ancestor as CYP79 enzymes, some of which also, in common with CYP73A5, use aromatic substrates derived from the shikimate pathway (2, 3). It was thus tempting to speculate that the substrate of CYP98 enzymes was derived from aromatic amino acids as well. The Arabidopsis CYP98 family is formed by only three genes. CYP98A3 is present in single copy; CYP98A8 and CYP98A9 are 69% identical to one another and only 52% identical to CYP98A3. All P450 genes in the phenylpropanoid pathway (CYP73A5, CYP84A1, and CYP...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

CYP98A3 from Arabidopsis thaliana Is a 3′-Hydroxylase of Phenolic Esters, a Missing Link in the Phenylpropanoid Pathway

Schoch

Goepfert

Morant

et al. 2001

Journal of Biological Chemistry

405

370

View full text Add to dashboard Cite

show abstract

“…4-Coumarate 3, which is formed from cinnamate 2 by the action of a highly substrate-specific P450 monooxygenase of the CYP73 family, cinnamate 4-hydroxylase (39 -41), is the most preferred substrate of 4-coumarate:CoA ligase (4CL), and 4-coumaroyl-CoA ester 4 the most important 4CL product (36 -38). 4-Coumaroyl-CoA 4 can then be converted to caffeoyl-CoA 6 by a 4-coumaroyl-CoA 3-hydroxylase (CCoA3H) such as that reported in Silene dioica (42), parsley (43), Zinnia (44), and Lithospermum erythrorhizon (45), although these activities may actually be polyphenol oxidases (45)(46)(47). Thus, the conversion of 4-coumarate 3 to caffeoyl-CoA 6 via 4-coumaroyl-CoA 4 represents one lignin precursor pathway.…”

Section: Discussionmentioning

confidence: 99%

5-Hydroxyconiferyl Aldehyde Modulates Enzymatic Methylation for Syringyl Monolignol Formation, a New View of Monolignol Biosynthesis in Angiosperms

Popko

Umezawa

et al. 2000

Journal of Biological Chemistry

231

195

View full text Add to dashboard Cite

S-Adenosyl-L-methionine-dependent caffeate O-methyltransferase (COMT, EC 2.1.1.6) has traditionally been thought to catalyze the methylation of caffeate and 5-hydroxyferulate for the biosynthesis of syringyl monolignol, a lignin constituent of angiosperm wood that enables efficient lignin degradation for cellulose production. However, recent recognition that coniferyl aldehyde prevents 5-hydroxyferulate biosynthesis in lignifying tissue, and that the hydroxylated form of coniferyl aldehyde, 5-hydroxyconiferyl aldehyde, is an alternative COMT substrate, demands a re-evaluation of the role of COMT during monolignol biosynthesis. Based on recombinant aspen (Populus tremuloides) COMT enzyme kinetics coupled with mass spectrometry analysis, this study establishes for the first time that COMT is in fact a 5-hydroxyconiferyl aldehyde O-methyltransferase (AldOMT), and that 5-hydroxyconiferyl aldehyde is both the preferred AldOMT substrate and an inhibitor of caffeate and 5-hydroxyferulate methylation, as measured by K m and K i values. 5-Hydroxyconiferyl aldehyde also inhibited the caffeate and 5-hydroxyferulate methylation activities of xylem proteins from various angiosperm tree species. The evidence that syringyl monolignol biosynthesis is independent of caffeate and 5-hydroxyferulate methylation supports our previous discovery that coniferyl aldehyde prevents ferulate 5-hydroxylation and at the same time ensures a coniferyl aldehyde 5-hydroxylase (CAld5H)-mediated biosynthesis of 5-hydroxyconiferyl aldehyde. Together, our results provide conclusive evidence for the presence of a CAld5H/AldOMT-catalyzed coniferyl aldehyde 5-hydroxylation/methylation pathway that directs syringyl monolignol biosynthesis in angiosperms.Regardless of its importance to tree growth, lignin is problematic to postharvest, cellulose-based wood processing for fiber, chemical, and energy production, because it must be degraded from cellulose at great expense. Certain structural constituents of lignin, such as the guaiacyl moiety, promote monomer cross-linkages that increase lignin resistance to degradation (1-3). In angiosperms, lignin is composed of a mixture of guaiacyl and syringyl monolignols, and can be degraded at considerably less energy and chemical cost than gymnosperm lignin, which consists almost entirely of guaiacyl moieties (4). As a result, there has been long standing incentive to understand the biosynthesis of syringyl monolignol in angiosperm trees in order to genetically engineer more syringyl lignin in trees to facilitate wood processing (5-8).For more than four decades, our thinking of syringyl monolignol biosynthesis in angiosperms has followed the doctrine that S-adenosyl-L-methionine-dependent caffeate O-methyltransferase (COMT) 1 -catalyzed methylation of caffeate and 5-hydroxyferulate constitutes a central pathway for the formation of syringyl monolignol via sinapate (Fig. 1A) . This methylation pathway has, however, been challenged, as the involvement in this pathway (Fig. 1A) of the ferulate 5-hydroxylation step (1...

show abstract

“…Nevertheless, NADPH and O 2 were required for their activity, and the enzymes were fully functional in the presence of reducing agents such as dithioerythritol (DTE; which was included in the extraction and assay buffers). This suggests that these hydroxylases in sweet basil peltate glands and leaves are not likely to be nonspecific phenolases (Patil and Zucker, 1965), nor are they likely to be the potential hydroxylases that have been reported to require FAD (Kamsteeg et al, 1981) or Zn 2ϩ and ascorbate (Kneusel et al, 1989) for activity. It seems likely that the enzyme(s) responsible for the 3Ј-hydroxylation of p-coumaroyl 4-hydroxyphenyllactate in sweet basil will be related to the analogous enzyme in C. blumei (Petersen et al, 1993;Petersen, 1997).…”

Section: Discussionmentioning

confidence: 99%

Differential Production of meta Hydroxylated Phenylpropanoids in Sweet Basil Peltate Glandular Trichomes and Leaves Is Controlled by the Activities of Specific Acyltransferases and Hydroxylases

et al. 2002

View full text Add to dashboard Cite

Sweet basil (Ocimum basilicum) peltate glandular trichomes produce a variety of small molecular weight phenylpropanoids, such as eugenol, caffeic acid, and rosmarinic acid, that result from meta hydroxylation reactions. Some basil lines do not synthesize eugenol but instead synthesize chavicol, a phenylpropanoid that does not contain a meta hydroxyl group. Two distinct acyltransferases, p-coumaroyl-coenzyme A:shikimic acid p-coumaroyl transferase and p-coumaroyl-coenzyme A:4-hydroxyphenyllactic acid p-coumaroyl transferase, responsible for the production of p-coumaroyl shikimate and of p-coumaroyl 4-hydroxyphenyllactate, respectively, were partially purified and shown to be specific for their substrates. p-Coumaroyl-coenzyme A:shikimic acid p-coumaroyl transferase is expressed in basil peltate glands that are actively producing eugenol and is not active in glands of noneugenol-producing basil plants, suggesting that the levels of this activity determine the levels of synthesis of some meta-hydroxylated phenylpropanoids in these glands such as eugenol. Two basil cDNAs encoding isozymes of cytochrome P450 CYP98A13, which meta hydroxylates p-coumaroyl shikimate, were isolated and found to be highly similar (90% identity) to the Arabidopsis homolog, CYP98A3. Like the Arabidopsis enzyme, the basil enzymes were found to be very specific for p-coumaroyl shikimate. Finally, additional hydroxylase activities were identified in basil peltate glands that convert p-coumaroyl 4-hydroxyphenyllactic acid to its caffeoyl derivative and p-coumaric acid to caffeic acid.Basil (Ocimum basilicum) plants synthesize significant quantities of phenylpropanoid derivatives that contain an hydroxyl group at the meta position on the aromatic ring. Several of these compounds, such as eugenol, rosmarinic acid, and caffeic acid (Fig. 1), are found in high concentrations in specialized structures that are located on the surface of the aerial parts of the plant and are known as peltate glandular trichomes (glands; Gang et al., 2001). These specialized metabolites have been found in other plant species as well, but the nature of the enzymes that catalyze the meta hydroxylation has so far remained poorly understood.One of the difficulties in identifying the enzymes catalyzing the 3-hydroxylations in the phenylpropanoid pathways is that they appear to be found at very low abundance in the tissues examined (Petersen, 1997). We have recently shown (Gang et al., 2001) that, as in peppermint (Mentha ϫ piperita; Gershenzon et al., 1992;McCaskill and Croteau, 1995), the basil gland cells can be removed from the plant and studied in isolation from the rest of the plant, greatly facilitating biochemical and molecular investigations of a single, fully differentiated cell type (Gershenzon et al., 1992;McCaskill et al., 1992;McCaskill and Croteau, 1995;Lange et al., 2000;Gang et al., 2001). Furthermore, because these gland cells are so highly specialized for production of plantspecialized metabolites, the enzymes in these metabolic pathways are highly express...

show abstract

Identification, Properties and Genetic Control of p-CoumaroylCoenzyme A, 3-Hydroxylase Isolated from Petals of Silene dioica

Cited by 43 publications

References 13 publications

CYP98A3 from Arabidopsis thaliana Is a 3′-Hydroxylase of Phenolic Esters, a Missing Link in the Phenylpropanoid Pathway

CYP98A3 from Arabidopsis thaliana Is a 3′-Hydroxylase of Phenolic Esters, a Missing Link in the Phenylpropanoid Pathway

5-Hydroxyconiferyl Aldehyde Modulates Enzymatic Methylation for Syringyl Monolignol Formation, a New View of Monolignol Biosynthesis in Angiosperms

Differential Production of meta Hydroxylated Phenylpropanoids in Sweet Basil Peltate Glandular Trichomes and Leaves Is Controlled by the Activities of Specific Acyltransferases and Hydroxylases

Contact Info

Product

Resources

About