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Literature summary for 1.1.1.195 extracted from

  • Patel, P.; Gupta, N.; Gaikwad, S.; Agrawal, D.C.; Khan, B.M.
    Leucaena sp. recombinant cinnamyl alcohol dehydrogenase: purification and physicochemical characterization (2014), Int. J. Biol. Macromol., 63, 254-260.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
DTT activates 25% at 1 mM Leucaena leucocephala

Cloned(Commentary)

Cloned (Comment) Organism
gene CAD, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) Leucaena leucocephala

General Stability

General Stability Organism
the enzyme is resistant to osmolytes, reducing agents and non-ionic detergents Leucaena leucocephala

Inhibitors

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 [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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Leucaena leucocephala B5AR58 cv. K-636
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
stem
-
Leucaena leucocephala
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms Comment Organism
CAD
-
Leucaena leucocephala
cinnamyl alcohol dehydrogenase
-
Leucaena leucocephala
LlCAD2
-
Leucaena leucocephala

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Leucaena leucocephala

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
stable up to, half-life is 5 h Leucaena leucocephala

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
aldehyde reduction Leucaena leucocephala
9
-
alcohol oxidation Leucaena leucocephala

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 9 stable at, half-life is 15 h Leucaena leucocephala

Cofactor

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

General Information Comment Organism
metabolism enzyme CAD catalyzes the final step in monolignol biosynthesis, leading to lignin formation in plants Leucaena leucocephala

kcat/KM [mM/s]

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