Activating Compound | Comment | Organism | Structure |
---|---|---|---|
DTT | activates 25% at 1 mM | Leucaena leucocephala |
Cloned (Comment) | Organism |
---|---|
gene CAD, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) | Leucaena leucocephala |
General Stability | Organism |
---|---|
the enzyme is resistant to osmolytes, reducing agents and non-ionic detergents | Leucaena leucocephala |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | inhibits 90% at 10 mM | Leucaena leucocephala | |
Cs+ | - |
Leucaena leucocephala | |
CTAB | inhibits 92% at 10 mM | Leucaena leucocephala | |
Cu2+ | - |
Leucaena leucocephala | |
diethyldicarbonate | 60% inhibition at 1 mM | Leucaena leucocephala | |
EDTA | inhibits 92% at 10 mM | Leucaena leucocephala | |
Hg2+ | strong inhibition | Leucaena leucocephala | |
Li+ | strong inhibition | Leucaena leucocephala | |
additional information | poor inhibition by Triton X-100 and Tween 80 at 1%, and by glucose at 20% | Leucaena leucocephala | |
Ni2+ | strong inhibition | Leucaena leucocephala | |
SDS | inhibits 95% at 1% | Leucaena leucocephala | |
sucrose | 10% inhibition at 20% | Leucaena leucocephala | |
Zn2+ | the enzyme is a Zn-metalloenzyme with 4 Zn2+ per dimer. The enzyme is inhibited in presence of externally supplemented Zn2+ ions, strong inhibition | Leucaena leucocephala |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0025 | - |
NADPH | pH 6.5, 40°C | Leucaena leucocephala | |
0.0028 | - |
sinapyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
0.0035 | - |
p-coumaryl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
0.0037 | - |
coniferyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
0.0057 | - |
NADP+ | pH 8.8, 40°C | Leucaena leucocephala | |
0.0059 | - |
cinnamaldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
0.0086 | - |
sinapyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
0.0096 | - |
coniferyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
0.0113 | - |
cinnamyl alcohol | pH 8.8, 40°C | Leucaena leucocephala |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | slight activating effect by KCl at 100 mM | Leucaena leucocephala | |
NaCl | activates 15% at 100 mM | Leucaena leucocephala | |
Zn2+ | 4 Zn2+ per homodimer, the enzyme is a Zn-metalloenzyme with 4 Zn2+ per dimer. The enzyme is inhibited in presence of externally supplemented Zn2+ ions | Leucaena leucocephala |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
2 * 40000, recombinant His-tagged enzyme, SDS-PAGE | Leucaena leucocephala |
80000 | - |
gel filtration, recombinant His-tagged enzyme | Leucaena leucocephala |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cinnamyl alcohol + NADP+ | Leucaena leucocephala | - |
cinnamaldehyde + NADPH + H+ | - |
r | |
coniferyl alcohol + NADP+ | Leucaena leucocephala | - |
coniferyl aldehyde + NADPH + H+ | - |
r | |
p-coumaryl alcohol + NADP+ | Leucaena leucocephala | - |
p-coumaryl aldehyde + NADPH + H+ | - |
r | |
sinapyl alcohol + NADP+ | Leucaena leucocephala | - |
sinapyl aldehyde + NADPH + H+ | - |
r | |
sinapyl aldehyde + NADPH + H+ | Leucaena leucocephala | - |
sinapyl alcohol + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leucaena leucocephala | B5AR58 | cv. K-636 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 65fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, anion exchange chromatography, and again gel filtration | Leucaena leucocephala |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
stem | - |
Leucaena leucocephala | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
37.8 | - |
purified recombinant His-tagged enzyme, pH 6.25, 25°C | Leucaena leucocephala |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cinnamaldehyde + NADPH + H+ | - |
Leucaena leucocephala | cinnamyl alcohol + NADP+ | - |
r | |
cinnamyl alcohol + NADP+ | - |
Leucaena leucocephala | cinnamaldehyde + NADPH + H+ | - |
r | |
coniferyl alcohol + NADP+ | - |
Leucaena leucocephala | coniferyl aldehyde + NADPH + H+ | - |
r | |
coniferyl aldehyde + NADPH + H+ | - |
Leucaena leucocephala | coniferyl alcohol + NADP+ | - |
r | |
additional information | the enzyme shows broad substrate specificity, LlCAD2 shows significant activity against cinnamyl substrates, but is almost inactive against aliphatic and other benzyl compounds. Possible involvement of histidine at the active site. Low activity with benzaldehyde, anisaldehyde, vanillin, and syringaldehyde. No or very poor activity with formaldehyde, acetaldehyde, propionaldehyde, and butryaldehyde | Leucaena leucocephala | ? | - |
? | |
p-coumaryl alcohol + NADP+ | - |
Leucaena leucocephala | p-coumaryl aldehyde + NADPH + H+ | - |
r | |
p-coumaryl aldehyde + NADPH + H+ | - |
Leucaena leucocephala | p-coumaryl alcohol + NADP+ | - |
r | |
sinapyl alcohol + NADP+ | - |
Leucaena leucocephala | sinapyl aldehyde + NADPH + H+ | - |
r | |
sinapyl aldehyde + NADPH + H+ | - |
Leucaena leucocephala | sinapyl alcohol + NADP+ | - |
r | |
sinapyl aldehyde + NADPH + H+ | preferred substrate | Leucaena leucocephala | sinapyl alcohol + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 40000, recombinant His-tagged enzyme, SDS-PAGE | Leucaena leucocephala |
More | the dimer shows 12 free and 16 totalcysteine residues. Two disulfide linkages can be deduced per dimer from the data, one intra-subunit linkage per subunit, and one non-interchain linkage | Leucaena leucocephala |
Synonyms | Comment | Organism |
---|---|---|
CAD | - |
Leucaena leucocephala |
cinnamyl alcohol dehydrogenase | - |
Leucaena leucocephala |
LlCAD2 | - |
Leucaena leucocephala |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Leucaena leucocephala |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
stable up to, half-life is 5 h | Leucaena leucocephala |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 3.7 | coniferyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
16 | - |
sinapyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
16.2 | - |
cinnamyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
16.3 | - |
coniferyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
18 | - |
cinnamaldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
22.5 | - |
p-coumaryl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
23.3 | - |
NADP+ | pH 8.8, 40°C | Leucaena leucocephala | |
28.8 | - |
NADPH | pH 6.5, 40°C | Leucaena leucocephala | |
32.2 | - |
sinapyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
aldehyde reduction | Leucaena leucocephala |
9 | - |
alcohol oxidation | Leucaena leucocephala |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 9 | stable at, half-life is 15 h | Leucaena leucocephala |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme is highly cofactor specific and is active only in presence of NADP(H), exhibiting insignificant activity with NAD(H) (up to 3 mM) | Leucaena leucocephala | |
NADP+ | - |
Leucaena leucocephala | |
NADPH | - |
Leucaena leucocephala |
General Information | Comment | Organism |
---|---|---|
metabolism | enzyme CAD catalyzes the final step in monolignol biosynthesis, leading to lignin formation in plants | Leucaena leucocephala |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1400 | - |
cinnamyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
1700 | - |
coniferyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
1900 | - |
sinapyl alcohol | pH 8.8, 40°C | Leucaena leucocephala | |
3100 | - |
cinnamaldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
4100 | - |
NADP+ | pH 8.8, 40°C | Leucaena leucocephala | |
6400 | - |
coniferyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
6400 | - |
p-coumaryl aldehyde | pH 6.5, 40°C | Leucaena leucocephala | |
11500 | - |
NADPH | pH 6.5, 40°C | Leucaena leucocephala | |
11600 | - |
sinapyl aldehyde | pH 6.5, 40°C | Leucaena leucocephala |