Cloned (Comment) | Organism |
---|---|
sequence comparisons, recombinant expression of MBP-tagged enzyme in Nicotiana tabacum cv. Nc89 leaves and in Fragaria x ananassa cv. Hongyan ripening fruits via transfection with Agrobacterium strain EHA105, quantitative PCR enzyme expression analysis, analysis of flavonoid contents in the transgenic plants | Paeonia suffruticosa |
sequence comparisons, recombinant expression of MBP-tagged enzyme in Nicotiana tabacum cv. Nc89 leaves and in Fragaria × ananassa cv. Hongyan ripening fruits via transfection with Agrobacterium strain EHA105, quantitative PCR enzyme expression analysis, analysis of flavonoid contents in the transgenic plants | Paeonia tenuifolia |
Protein Variants | Comment | Organism |
---|---|---|
G13E | site-directed mutagenesis, the mutant shows increased activity compared to wild-type | Paeonia tenuifolia |
L87A | site-directed mutagenesis, the mutant shows highly decreased catalytic efficency, the observed in vitro catalytic efficiency of PtAOMT-L87R mutant is equal to that of Paeonia tenuifolia PtAOMT wild-type | Paeonia suffruticosa |
L87R | site-directed mutagenesis, the mutant shows highly decreased catalytic efficency, the observed in vitro catalytic efficiency of PtAOMT-L87R mutant is equal to that of Paeonia tenuifolia PtAOMT wild-type | Paeonia suffruticosa |
R87L | site-directed mutagenesis, the mutant shows highly increased catalytic efficency, the observed in vitro catalytic efficiency of PtAOMT-R87L mutant is equal to that of Paeonia suffruticosa PsAOMT wild-type | Paeonia tenuifolia |
T205R | site-directed mutagenesis, the mutant shows increased activity compared to wild-type | Paeonia tenuifolia |
T85A | site-directed mutagenesis, the mutant shows increased activity compared to wild-type | Paeonia tenuifolia |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00106 | - |
cyanidin 3,5-O-diglucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.00176 | - |
cyanidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.00411 | - |
delphinidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.00433 | - |
quercetin | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.0123 | - |
quercetin 3-O-rutinoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.147 | - |
luteolin 7-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates, less effective compared to Mg2+ | Paeonia suffruticosa | |
Co2+ | activates, less effective compared to Mg2+ | Paeonia suffruticosa | |
Mg2+ | best activating cation, optimal at 1 mM | Paeonia suffruticosa | |
Mn2+ | activates, less effective compared to Mg2+ | Paeonia suffruticosa | |
Zn2+ | activates, less effective compared to Mg2+ | Paeonia suffruticosa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + a 3'-hydroxyflavonoid | Paeonia suffruticosa | - |
S-adenosyl-L-homocysteine + a 3'-methoxyflavonoid | - |
? | |
S-adenosyl-L-methionine + a 3'-hydroxyflavonoid | Paeonia tenuifolia | - |
S-adenosyl-L-homocysteine + a 3'-methoxyflavonoid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paeonia suffruticosa | - |
cv. Gunpohden | - |
Paeonia tenuifolia | - |
- |
- |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
flower | - |
Paeonia suffruticosa | - |
flower | - |
Paeonia tenuifolia | - |
leaf | - |
Paeonia suffruticosa | - |
leaf | low level | Paeonia tenuifolia | - |
additional information | developmental expression of enzyme PsAOMT, and determination of flavonoid profile, overview | Paeonia suffruticosa | - |
additional information | developmental expression of enzyme PtAOMT, and determination of flavonoid profile, overview | Paeonia tenuifolia | - |
petal | - |
Paeonia suffruticosa | - |
petal | - |
Paeonia tenuifolia | - |
root | low level | Paeonia suffruticosa | - |
root | low level | Paeonia tenuifolia | - |
sepal | - |
Paeonia suffruticosa | - |
sepal | - |
Paeonia tenuifolia | - |
stem | - |
Paeonia tenuifolia | - |
stem | low level | Paeonia suffruticosa | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.000003 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate luteolin | Paeonia suffruticosa |
0.000024 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate cyanidin | Paeonia suffruticosa |
0.000125 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate luteolin | Paeonia tenuifolia |
0.000134 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate luteolin 7-O-glucoside | Paeonia tenuifolia |
0.00105 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate quercetin | Paeonia tenuifolia |
0.0013 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate delphinidin 3-O-glucoside | Paeonia tenuifolia |
0.00178 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate cyanidin 3,5-di-O-glucoside | Paeonia tenuifolia |
0.00181 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate quercetin 3-O-rutinoside | Paeonia tenuifolia |
0.0023 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate cyanidin 3-O-glucoside | Paeonia tenuifolia |
0.082 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate delphinidin 3-O-glucoside | Paeonia suffruticosa |
0.085 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate quercetin | Paeonia suffruticosa |
0.107 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate cyanidin 3,5-O-diglucoside | Paeonia suffruticosa |
0.121 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate luteolin 7-O-glucoside | Paeonia suffruticosa |
0.139 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate cyanidin 3-O-glucoside | Paeonia suffruticosa |
0.165 | - |
recombinant enzyme, pH 7.5, temperature not specified in the publication, substrate quercetin 3-O-rutinoside | Paeonia suffruticosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | substrate specificity study using a number of potential substrates including anthocyanidins, anthocyanins, flavonols, flavones, flavan-3-ols, and phenolic acid as substrates based on optimized conditions, overview. Enzyme PsAOMT uses anthocyanins as methoxyl accepters, and acts to methylate the 3'-hydroxyl group of the B-ring with high affinity and efficiency. Pelargonidin 3-O-glucoside is the only tested anthocyanin compound that is not a substrate for PsAOMT. With delphinidin 3-O-glucoside, a sequential methylation occurs at the 3'- and 5'-hydroxyl group on the B-ring. In comparison with the substrates delphinidin 3-O-glucoside, quercetin 3-O-rutinoside, and quercetin, PsAOMT has a higher affinity for cyanidin 3-O-glucoside and cyanidin 3,5-O-diglucoside. Cyanidin-derived anthocyanins are high-affinity substrates for PsAOMT. No activity with the antocyanin pelargonidin 3-O-glucoside and the cyanidin delphinin. The enzyme also shows no activity with kaempferol 3-O-glucoside, kaempferol, apigenin, naringenin, epicatechin, and caffeic acid | Paeonia suffruticosa | ? | - |
- |
|
additional information | substrate specificity study using a number of potential substrates including anthocyanidins, anthocyanins, flavonols, flavones, flavan-3-ols, and phenolic acid as substrates based on optimized conditions, overview. Enzyme PtAOMT uses anthocyanins as methoxyl accepters, and acts to methylate the 3'-hydroxyl group of the B-ring with high affinity and efficiency. Pelargonidin 3-O-glucoside is the only tested anthocyanin compound that is not a substrate for PtAOMT. With delphinidin 3-O-glucoside, a sequential methylation occurs at the 3'- and 5'-hydroxyl group on the B-ring. In comparison with the substrates delphinidin 3-O-glucoside, quercetin 3-O-rutinoside, and quercetin, PtAOMT has a higher affinity for cyanidin 3-O-glucoside and cyanidin 3,5-O-diglucoside. Cyanidin-derived anthocyanins are high-affinity substrates for PtAOMT. No activity with the antocyanin pelargonidin 3-O-glucoside and the cyanidin delphinin. The enzyme also shows no activity with kaempferol 3-O-glucoside, kaempferol, apigenin, naringenin, epicatechin, and caffeic acid | Paeonia tenuifolia | ? | - |
- |
|
S-adenosyl-L-methionine + 3'-methoxydelphinidin 3-O-glucoside | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3',5'-dimethoxydelphinidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + 3'-methoxydelphinidin 3-O-glucoside | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3',5'-dimethoxydelphinidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + a 3'-hydroxyflavonoid | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + a 3'-methoxyflavonoid | - |
? | |
S-adenosyl-L-methionine + a 3'-hydroxyflavonoid | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + a 3'-methoxyflavonoid | - |
? | |
S-adenosyl-L-methionine + cyanidin | very low activity | Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxycyanidin | - |
? | |
S-adenosyl-L-methionine + cyanidin 3,5-O-diglucoside | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxycyanidin 3,5-O-diglucoside | - |
? | |
S-adenosyl-L-methionine + cyanidin 3,5-O-diglucoside | substrate binding structure, modelling, overview | Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxycyanidin 3,5-O-diglucoside | - |
? | |
S-adenosyl-L-methionine + cyanidin 3-O-glucoside | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxycyanidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + cyanidin 3-O-glucoside | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxycyanidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + delphinidin 3-O-glucoside | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxydelphinidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + delphinidin 3-O-glucoside | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxydelphinidin 3-O-glucoside | - |
? | |
S-adenosyl-L-methionine + luteolin | very low activity | Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxyluteolin | - |
? | |
S-adenosyl-L-methionine + luteolin | low activity | Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxyluteolin | - |
? | |
S-adenosyl-L-methionine + luteolin 7-O-glucoside | low activity | Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxyluteolin 7-O-glucoside | - |
? | |
S-adenosyl-L-methionine + luteolin 7-O-glucoside | low activity | Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxyluteolin 7-O-glucoside | - |
? | |
S-adenosyl-L-methionine + quercetin | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxyquercetin | - |
? | |
S-adenosyl-L-methionine + quercetin | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxyquercetin | - |
? | |
S-adenosyl-L-methionine + quercetin 3-O-rutinoside | - |
Paeonia suffruticosa | S-adenosyl-L-homocysteine + 3'-methoxyquercetin 3-O-rutinoside | - |
? | |
S-adenosyl-L-methionine + quercetin 3-O-rutinoside | - |
Paeonia tenuifolia | S-adenosyl-L-homocysteine + 3'-methoxyquercetin 3-O-rutinoside | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 66000, recombinant of MBP-tagged enzyme, SDS-PAGE, x * 26000, about, native enzyme, sequence calculation | Paeonia suffruticosa |
? | x * 66000, recombinant of MBP-tagged enzyme, SDS-PAGE, x * 26000, about, native enzyme, sequence calculation | Paeonia tenuifolia |
Synonyms | Comment | Organism |
---|---|---|
anthocyanin O-methyltransferase | - |
Paeonia suffruticosa |
anthocyanin O-methyltransferase | - |
Paeonia tenuifolia |
AOMT | - |
Paeonia suffruticosa |
AOMT | - |
Paeonia tenuifolia |
PsAOMT | - |
Paeonia suffruticosa |
PtAOMT | - |
Paeonia tenuifolia |
type II OMT | - |
Paeonia suffruticosa |
type II OMT | - |
Paeonia tenuifolia |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00022 | - |
luteolin 7-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.00983 | - |
quercetin | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.0947 | - |
delphinidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.128 | - |
cyanidin 3,5-O-diglucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.162 | - |
cyanidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
0.192 | - |
quercetin 3-O-rutinoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
recombinant enzyme | Paeonia suffruticosa |
7.5 | - |
recombinant enzyme | Paeonia tenuifolia |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8.5 | activity range, recombinant enzyme | Paeonia suffruticosa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Paeonia suffruticosa | |
S-adenosyl-L-methionine | - |
Paeonia tenuifolia |
General Information | Comment | Organism |
---|---|---|
evolution | the catalytic activity of Paeonia tenuifolia PtAOMT is much lower than that of Paeonia suffruticosa PsAOMT (approximately 60fold lower), the two enzymes show identical substrate specificity | Paeonia suffruticosa |
evolution | the catalytic activity of Paeonia tenuifolia PtAOMT is much lower than that of Paeonia suffruticosa PsAOMT (approximately 60fold lower), the two enzymes show identical substrate specificity | Paeonia tenuifolia |
additional information | enzyme structure homology modeling using the three-domensional structure of caffeoyl-CoA 3-O-methyltransferase (CCoAOMT, PDB ID 1SUI) from alfalfa, substrate binding sites are predicted by docking Cy3G5G with the PsAOMT dimer 3D model. Molecular model of the PsAOMT active site. Structure-function analysis | Paeonia suffruticosa |
additional information | enzyme structure homology modeling using the three-domensional structure of caffeoyl-CoA 3-O-methyltransferase (CCoAOMT, PDB ID 1SUI) from alfalfa. Structure-function analysis | Paeonia tenuifolia |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0015 | - |
luteolin 7-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
2.27 | - |
quercetin | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
15.61 | - |
quercetin 3-O-rutinoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
23.1 | - |
delphinidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
92.045 | - |
cyanidin 3-O-glucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa | |
120.75 | - |
cyanidin 3,5-O-diglucoside | recombinant enzyme, pH 7.5, temperature not specified in the publication | Paeonia suffruticosa |