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Literature summary extracted from

  • Bhattacharyya, D.; Hazra, S.; Banerjee, A.; Datta, R.; Kumar, D.; Chakrabarti, S.; Chattopadhyay, S.
    Transcriptome-wide identification and characterization of CAD isoforms specific for podophyllotoxin biosynthesis from Podophyllum hexandrum (2016), Plant Mol. Biol., 92, 1-23.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.195 expressed in Escherichia coli BL21(DE3) cells Sinopodophyllum hexandrum
1.1.1.195 gene PhCAD1, DNA and amino acid sequence determination and analysis, genotyping and phylogenetic analysis and transcriptome analysis, semi-quantitative RT-PCR enzyme expression analysis, overview. Recombinant expression of His-tagged enzyme Sinopodophyllum hexandrum
1.1.1.195 gene PhCAD2, DNA and amino acid sequence determination and analysis, genotyping and phylogenetic analysis and transcriptome analysis, semi-quantitative RT-PCR enzyme expression analysis, overview. Recombinant expression of His-tagged enzyme Sinopodophyllum hexandrum
1.1.1.195 gene PhCAD3, DNA and amino acid sequence determination and analysis, genotyping and phylogenetic analysis and transcriptome analysis, semi-quantitative RT-PCR enzyme expression analysis, overview. Recombinant expression of His-tagged enzyme Sinopodophyllum hexandrum
1.1.1.195 gene PhCAD4, DNA and amino acid sequence determination and analysis, genotyping and phylogenetic analysis and transcriptome analysis, semi-quantitative RT-PCR enzyme expression analysis, overview. Recombinant expression of His-tagged enzyme Sinopodophyllum hexandrum

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.195 additional information in PhCAD1 the active site residue G302 from the sinapyl alcohol dehydrogenase, SAD, is mutated to C302 and active site residue A293 is mutated to M293. Accumulation of ptox and lignin in PhCAD1-4 isoforms overexpressing transgenic lines, overview Sinopodophyllum hexandrum
1.1.1.195 additional information in PhCAD2 the active site residue G302 from the sinapyl alcohol dehydrogenase, SAD, is mutated to A300 and active site residue A293 is mutated to D290 Sinopodophyllum hexandrum
1.1.1.195 additional information in PhCAD3 the active site residue G302 from the sinapyl alcohol dehydrogenase, SAD, is conserved, but active site residue A293 is mutated to T290. Accumulation of ptox and lignin in PhCAD1-4 isoforms overexpressing transgenic lines, overview Sinopodophyllum hexandrum
1.1.1.195 additional information in PhCAD4 the active site residue G302 from the sinapyl alcohol dehydrogenase, SAD, is conserved, but active site residue A293 is mutated to M296 Sinopodophyllum hexandrum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.195 0.0369
-
 coniferaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.04792
-
 sinapaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.0676
-
 coniferaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.0797
-
 coniferaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.1136
-
 coniferaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.1378
-
 sinapaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.1699
-
 sinapaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 0.2082
-
 sinapaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.195 Zn2+ required Sinopodophyllum hexandrum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.195 38700
-
-
 Sinopodophyllum hexandrum
1.1.1.195 38800
-
-
 Sinopodophyllum hexandrum
1.1.1.195 38900
-
-
 Sinopodophyllum hexandrum
1.1.1.195 42100
-
-
 Sinopodophyllum hexandrum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.195 cinnamaldehyde + NADPH + H+ Sinopodophyllum hexandrum
-
 cinnamyl alcohol + NADP+
-
 ?
1.1.1.195 coniferaldehyde + NADPH + H+ Sinopodophyllum hexandrum isoform CAD3 has a higher binding preference with coniferaldehyde over sinapaldehyde, followed by isoforms CAD4, CAD2, and CAD1, respectively coniferyl alcohol + NADP+
-
 ?
1.1.1.195 coniferyl aldehyde + NADPH + H+ Sinopodophyllum hexandrum
-
 coniferyl alcohol + NADP+
-
 r
1.1.1.195 sinapaldehyde + NADPH + H+ Sinopodophyllum hexandrum
-
 sinapyl alcohol + NADP+
-
 ?
1.1.1.195 sinapyl aldehyde + NADPH + H+ Sinopodophyllum hexandrum
-
 sinapyl alcohol + NADP+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.195 Sinopodophyllum hexandrum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.195 Ni-NTA column chromatography Sinopodophyllum hexandrum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.195 callus from leaves Sinopodophyllum hexandrum
-
1.1.1.195 cell suspension culture
-
 Sinopodophyllum hexandrum
-
1.1.1.195 leaf
-
 Sinopodophyllum hexandrum
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.195 cinnamaldehyde + NADPH + H+
-
 Sinopodophyllum hexandrum cinnamyl alcohol + NADP+
-
 ?
1.1.1.195 coniferaldehyde + NADPH + H+ isoform CAD3 has a higher binding preference with coniferaldehyde over sinapaldehyde, followed by isoforms CAD4, CAD2, and CAD1, respectively Sinopodophyllum hexandrum coniferyl alcohol + NADP+
-
 ?
1.1.1.195 coniferyl aldehyde + NADPH + H+
-
 Sinopodophyllum hexandrum coniferyl alcohol + NADP+
-
 r
1.1.1.195 sinapaldehyde + NADPH + H+
-
 Sinopodophyllum hexandrum sinapyl alcohol + NADP+
-
 ?
1.1.1.195 sinapyl aldehyde + NADPH + H+
-
 Sinopodophyllum hexandrum sinapyl alcohol + NADP+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.195 ? x * 38700, isoform CAD4, calculated from amino acid sequence Sinopodophyllum hexandrum
1.1.1.195 ? x * 38800, isoform CAD4, calculated from amino acid sequence Sinopodophyllum hexandrum
1.1.1.195 ? x * 38900, isoform CAD4, calculated from amino acid sequence Sinopodophyllum hexandrum
1.1.1.195 ? x * 42100, isoform CAD4, calculated from amino acid sequence Sinopodophyllum hexandrum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.195 CAD
-
 Sinopodophyllum hexandrum
1.1.1.195 CAD1 isoform Sinopodophyllum hexandrum
1.1.1.195 CAD2 isoform Sinopodophyllum hexandrum
1.1.1.195 CAD3 isoform Sinopodophyllum hexandrum
1.1.1.195 CAD4 isoform Sinopodophyllum hexandrum
1.1.1.195 cinnamyl alcohol dehydrogenase
-
 Sinopodophyllum hexandrum
1.1.1.195 PhCAD1
-
 Sinopodophyllum hexandrum
1.1.1.195 PhCAD2
-
 Sinopodophyllum hexandrum
1.1.1.195 PhCAD3
-
 Sinopodophyllum hexandrum
1.1.1.195 PhCAD4
-
 Sinopodophyllum hexandrum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.195 30
-
-
 Sinopodophyllum hexandrum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.195 2.4
-
 sinapaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 2.76
-
 sinapaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 3.4
-
 sinapaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 7.9
-
 coniferaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 8.3
-
 sinapaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 9
-
 coniferaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 10.8
-
 coniferaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 12
-
 coniferaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.195 7.5
-
-
 Sinopodophyllum hexandrum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.195 NADP+
-
 Sinopodophyllum hexandrum
1.1.1.195 NADPH
-
 Sinopodophyllum hexandrum

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.195 Sinopodophyllum hexandrum isoform CAD4, calculated from amino acid sequence
-
 5.9
1.1.1.195 Sinopodophyllum hexandrum isoform CAD3, calculated from amino acid sequence
-
 6
1.1.1.195 Sinopodophyllum hexandrum isoform CAD2, calculated from amino acid sequence
-
 6.2
1.1.1.195 Sinopodophyllum hexandrum isoform CAD1, calculated from amino acid sequence
-
 6.5

Expression

EC Number Organism Comment Expression
1.1.1.195 Sinopodophyllum hexandrum the CAD genes are strongly upregulated by methyljasmonate, transcriptome analysis, overview up
1.1.1.195 Sinopodophyllum hexandrum the CAD genes are strongly upregulated by methyljasmonate, transcriptome analysis, overview. Accumulation of ptox and lignin in PhCAD1-4 isoforms overexpressing transgenic lines, overview up

General Information

EC Number General Information Comment Organism
1.1.1.195 additional information enzyme molecular docking analysis with substrates, active site structure, structure comparisons between isozymes, overview Sinopodophyllum hexandrum
1.1.1.195 physiological function CAD catalyzes the synthesis of coniferyl alcohol and sinapyl alcohol from coniferaldehyde (CAld) and sinapaldehyde respectively. Coniferyl alcohol can produce both lignin and lignan while sinapyl alcohol produces only lignin. Accumulation of ptox and lignin in PhCAD1-4 isoforms overexpressing transgenic lines, overview. Podophyllotoxin (ptox) is a therapeutically important lignan with economic importance derived from Podophyllum hexandrum and is used as a precursor for the synthesis of anticancer drugs etoposide, teniposide and etopophose Sinopodophyllum hexandrum

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 11
-
 sinapaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 16
-
 sinapaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 24
-
 sinapaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 70
-
 coniferaldehyde isoform CAD2, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 130
-
 coniferaldehyde isoform CAD4, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 170
-
 sinapaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 180
-
 coniferaldehyde isoform CAD3, at pH 7.5 and 30°C Sinopodophyllum hexandrum
1.1.1.195 210
-
 coniferaldehyde isoform CAD1, at pH 7.5 and 30°C Sinopodophyllum hexandrum