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Literature summary 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

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

Cloned(Commentary)

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

General Stability

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.195 1,10-phenanthroline inhibits 90% at 10 mM Leucaena leucocephala
1.1.1.195 Cs+
-
 Leucaena leucocephala
1.1.1.195 CTAB inhibits 92% at 10 mM Leucaena leucocephala
1.1.1.195 Cu2+
-
 Leucaena leucocephala
1.1.1.195 diethyldicarbonate 60% inhibition at 1 mM Leucaena leucocephala
1.1.1.195 EDTA inhibits 92% at 10 mM Leucaena leucocephala
1.1.1.195 Hg2+ strong inhibition Leucaena leucocephala
1.1.1.195 Li+ strong inhibition Leucaena leucocephala
1.1.1.195 additional information poor inhibition by Triton X-100 and Tween 80 at 1%, and by glucose at 20% Leucaena leucocephala
1.1.1.195 Ni2+ strong inhibition Leucaena leucocephala
1.1.1.195 SDS inhibits 95% at 1% Leucaena leucocephala
1.1.1.195 sucrose 10% inhibition at 20% Leucaena leucocephala
1.1.1.195 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.195 0.0025
-
 NADPH pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 0.0028
-
 sinapyl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 0.0035
-
 p-coumaryl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 0.0037
-
 coniferyl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 0.0057
-
 NADP+ pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 0.0059
-
 cinnamaldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 0.0086
-
 sinapyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 0.0096
-
 coniferyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 0.0113
-
 cinnamyl alcohol pH 8.8, 40°C Leucaena leucocephala

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.195 additional information slight activating effect by KCl at 100 mM Leucaena leucocephala
1.1.1.195 NaCl activates 15% at 100 mM Leucaena leucocephala
1.1.1.195 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]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.195 40000
-
 2 * 40000, recombinant His-tagged enzyme, SDS-PAGE Leucaena leucocephala
1.1.1.195 80000
-
 gel filtration, recombinant His-tagged enzyme Leucaena leucocephala

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.195 cinnamyl alcohol + NADP+ Leucaena leucocephala
-
 cinnamaldehyde + NADPH + H+
-
 r
1.1.1.195 coniferyl alcohol + NADP+ Leucaena leucocephala
-
 coniferyl aldehyde + NADPH + H+
-
 r
1.1.1.195 p-coumaryl alcohol + NADP+ Leucaena leucocephala
-
 p-coumaryl aldehyde + NADPH + H+
-
 r
1.1.1.195 sinapyl alcohol + NADP+ Leucaena leucocephala
-
 sinapyl aldehyde + NADPH + H+
-
 r
1.1.1.195 sinapyl aldehyde + NADPH + H+ Leucaena leucocephala
-
 sinapyl alcohol + NADP+
-
 r

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.195 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

EC Number Source Tissue Comment Organism Textmining
1.1.1.195 stem
-
 Leucaena leucocephala
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.195 37.8
-
 purified recombinant His-tagged enzyme, pH 6.25, 25°C Leucaena leucocephala

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.195 cinnamaldehyde + NADPH + H+
-
 Leucaena leucocephala cinnamyl alcohol + NADP+
-
 r
1.1.1.195 cinnamyl alcohol + NADP+
-
 Leucaena leucocephala cinnamaldehyde + NADPH + H+
-
 r
1.1.1.195 coniferyl alcohol + NADP+
-
 Leucaena leucocephala coniferyl aldehyde + NADPH + H+
-
 r
1.1.1.195 coniferyl aldehyde + NADPH + H+
-
 Leucaena leucocephala coniferyl alcohol + NADP+
-
 r
1.1.1.195 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 ?
-
 ?
1.1.1.195 p-coumaryl alcohol + NADP+
-
 Leucaena leucocephala p-coumaryl aldehyde + NADPH + H+
-
 r
1.1.1.195 p-coumaryl aldehyde + NADPH + H+
-
 Leucaena leucocephala p-coumaryl alcohol + NADP+
-
 r
1.1.1.195 sinapyl alcohol + NADP+
-
 Leucaena leucocephala sinapyl aldehyde + NADPH + H+
-
 r
1.1.1.195 sinapyl aldehyde + NADPH + H+
-
 Leucaena leucocephala sinapyl alcohol + NADP+
-
 r
1.1.1.195 sinapyl aldehyde + NADPH + H+ preferred substrate Leucaena leucocephala sinapyl alcohol + NADP+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.195 homodimer 2 * 40000, recombinant His-tagged enzyme, SDS-PAGE Leucaena leucocephala
1.1.1.195 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

EC Number Synonyms Comment Organism
1.1.1.195 CAD
-
 Leucaena leucocephala
1.1.1.195 cinnamyl alcohol dehydrogenase
-
 Leucaena leucocephala
1.1.1.195 LlCAD2
-
 Leucaena leucocephala

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.195 40
-
-
 Leucaena leucocephala

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.195 40
-
 stable up to, half-life is 5 h Leucaena leucocephala

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.195 2 3.7 coniferyl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 16
-
 sinapyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 16.2
-
 cinnamyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 16.3
-
 coniferyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 18
-
 cinnamaldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 22.5
-
 p-coumaryl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 23.3
-
 NADP+ pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 28.8
-
 NADPH pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 32.2
-
 sinapyl aldehyde pH 6.5, 40°C Leucaena leucocephala

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.195 6.5
-
 aldehyde reduction Leucaena leucocephala
1.1.1.195 9
-
 alcohol oxidation Leucaena leucocephala

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.195 7 9 stable at, half-life is 15 h Leucaena leucocephala

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.195 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
1.1.1.195 NADP+
-
 Leucaena leucocephala
1.1.1.195 NADPH
-
 Leucaena leucocephala

General Information

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

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.195 1400
-
 cinnamyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 1700
-
 coniferyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 1900
-
 sinapyl alcohol pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 3100
-
 cinnamaldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 4100
-
 NADP+ pH 8.8, 40°C Leucaena leucocephala
1.1.1.195 6400
-
 coniferyl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 6400
-
 p-coumaryl aldehyde pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 11500
-
 NADPH pH 6.5, 40°C Leucaena leucocephala
1.1.1.195 11600
-
 sinapyl aldehyde pH 6.5, 40°C Leucaena leucocephala