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Literature summary for 1.1.1.195 extracted from

  • Jun, S.Y.; Walker, A.M.; Kim, H.; Ralph, J.; Vermerris, W.; Sattler, S.E.; Kang, C.
    The enzyme activity and substrate specificity of two major cinnamyl alcohol dehydrogenases in sorghum (Sorghum bicolor), SbCAD2 and SbCAD4 (2017), Plant Physiol., 174, 2128-2145 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.1.195

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta cells Sorghum bicolor

Crystallization (Commentary)

Crystallization (Comment) Organism
isoform CAD4, hanging drop vapor diffusion method, using 2% (v/v) tacsimate, pH 7.0, 5% (v/v) 2-propanol, 0.1 M imidazole, pH 7.0, and 8% (w/v) polyethylene glycol 3350 Sorghum bicolor

Protein Variants

Protein Variants Comment Organism
A278V the isoform CAD4 mutant shows strongly decreased catalytic efficiency compared to the wild type enzyme Sorghum bicolor
G301F the isoform CAD4 mutant enzyme displays slightly elevated Km values for 4-coumaryl aldehyde but decreased Km for coniferyl aldehyde compared with those values of wild type. The mutant becomes more dedicated toward sinapyl aldehyde, with substantially decreased efficiency for coniferyl aldehyde compared to the wild type enzyme Sorghum bicolor
L119W the isoform CAD4 mutant displays a slightly reduced Km value for both 4-coumaryl aldehyde and coniferyl aldehyde compared to the wild type enzyme. The mutant becomes more efficient toward coniferaldehyde due to the very small Km value, with substantially decreased efficiency for both sinapyl aldehyde and 4-coumaryl aldehyde Sorghum bicolor
L119W/G301F the isoform CAD4 double mutant displays its substrate preference in the order coniferyl aldehyde over 4-coumaryl aldehyde over sinapyl aldehyde, with higher catalytic efficiency than that of wild type enzyme. The mutant displays 10 and 800fold increases in its catalytic efficiency for coniferyl aldehyde and 4-coumaryl aldehyde, respectively, and a 50% decrease in catalytic efficiency for sinapyl aldehyde as compared to the wild type enzyme Sorghum bicolor
W58L the isoform CAD4 mutant enzyme displays slightly elevated Km values for 4-coumaryl aldehyde but decreased Km for coniferyl aldehyde compared with those values of wild type Sorghum bicolor
Y288P the isoform CAD4 mutant becomes more dedicated toward sinapyl aldehyde, with substantially decreased efficiency for coniferyl aldehyde compared to the wild type enzyme Sorghum bicolor
Y95V the isoform CAD4 mutant shows decreased catalytic efficiency compared to the wild type enzyme Sorghum bicolor

Inhibitors

Inhibitors Comment Organism Structure
4-coumaryl aldehyde
-
 Sorghum bicolor
coniferyl aldehyde
-
 Sorghum bicolor
sinapyl aldehyde
-
 Sorghum bicolor

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0078
-
 sinapyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
0.0104
-
 coniferyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
0.0168
-
 sinapyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
0.0214
-
 coniferyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
0.028
-
 coniferyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
0.033
-
 sinapyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
0.05976
-
 sinapyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
0.068
-
 coniferyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
0.06975
-
 4-coumaryl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
0.0783
-
 sinapyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
0.0793
-
 sinapyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
0.0809
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
0.09518
-
 coniferyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
0.105
-
 sinapyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
0.112
-
 sinapyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
0.233
-
 coniferyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
0.268
-
 4-coumaryl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
0.387
-
 coniferyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
2.12
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
4.18
-
 4-coumaryl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
4.63
-
 4-coumaryl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
5.52
-
 sinapyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
5.68
-
 coniferyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
6.88
-
 4-coumaryl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
7.1
-
 4-coumaryl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
7.97
-
 4-coumaryl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
9.17
-
 coniferyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ dependent on Sorghum bicolor

Organism

Organism UniProt Comment Textmining
Sorghum bicolor A1ILL4
-
-
Sorghum bicolor C5XC49
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography and Mono Q column chromatography Sorghum bicolor

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-coumaryl aldehyde + NADPH + H+
-
 Sorghum bicolor 4-coumaryl alcohol + NADP+
-
 ?
coniferyl aldehyde + NADPH + H+
-
 Sorghum bicolor coniferyl alcohol + NADP+ ? ?
sinapyl aldehyde + NADPH + H+
-
 Sorghum bicolor sinapyl alcohol + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
CAD2 isoform Sorghum bicolor
CAD4 isoform Sorghum bicolor
cinnamyl alcohol dehydrogenase
-
 Sorghum bicolor

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.76
-
 coniferyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
1.05
-
 sinapyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
1.08
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
1.19
-
 4-coumaryl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
1.59
-
 sinapyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
2.17
-
 4-coumaryl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
2.33
-
 coniferyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
2.5
-
 sinapyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
3.03
-
 4-coumaryl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
3.27
-
 4-coumaryl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
3.32
-
 sinapyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
3.59
-
 4-coumaryl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
3.68
-
 coniferyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
3.76
-
 coniferyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
5.29
-
 sinapyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
9.26
-
 coniferyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
9.5
-
 4-coumaryl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
11.93
-
 4-coumaryl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
12.54
-
 sinapyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
12.77
-
 coniferyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
18.98
-
 sinapyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
20.28
-
 coniferyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
25.83
-
 coniferyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
30.16
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
31.75
-
 sinapyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
39.97
-
 sinapyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
71.57
-
 coniferyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor

Cofactor

Cofactor Comment Organism Structure
NADPH
-
 Sorghum bicolor

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0121
-
 coniferyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
0.0162
-
 coniferyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
0.0374
-
 sinapyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
0.083
-
 sinapyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
0.099
-
 4-coumaryl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
0.118
-
 sinapyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
0.123
-
 coniferyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
0.16
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
0.295
-
 coniferyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
0.295
-
 coniferyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
0.427
-
 coniferyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
0.711
-
 coniferyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
1.056
-
 sinapyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
1.23
-
 sinapyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
1.39
-
 sinapyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
1.455
-
 sinapyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
2.91
-
 sinapyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
4.17
-
 coniferyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
6.03
-
 coniferyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
7.63
-
 4-coumaryl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
10.02
-
 4-coumaryl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
20.59
-
 4-coumaryl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
31.92
-
 4-coumaryl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
32.67
-
 4-coumaryl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
68.08
-
 sinapyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.28
-
 4-coumaryl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
0.38
-
 4-coumaryl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
0.4
-
 coniferyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
0.45
-
 sinapyl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
0.46
-
 4-coumaryl aldehyde isoform CAD4 mutant W58L, at pH 6.5 and 30°C Sorghum bicolor
0.47
-
 4-coumaryl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
0.51
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
0.52
-
 4-coumaryl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
10
-
 sinapyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
10
-
 coniferyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
10
-
 coniferyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
40
-
 coniferyl aldehyde isoform CAD4 mutant A278V, at pH 6.5 and 30°C Sorghum bicolor
40
-
 4-coumaryl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
40
-
 sinapyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
90
-
 coniferyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
130
-
 coniferyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
140
-
 coniferyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
170
-
 4-coumaryl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
180
-
 sinapyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
200
-
 sinapyl aldehyde isoform CAD4 mutant Y288P, at pH 6.5 and 30°C Sorghum bicolor
220
-
 coniferyl aldehyde isoform CAD4 mutant L119W, at pH 6.5 and 30°C Sorghum bicolor
310
-
 sinapyl aldehyde isoform CAD4 mutant G301F, at pH 6.5 and 30°C Sorghum bicolor
360
-
 sinapyl aldehyde isoform CAD4 mutant Y95V, at pH 6.5 and 30°C Sorghum bicolor
370
-
 4-coumaryl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor
380
-
 sinapyl aldehyde wild type isoform CAD4, at pH 6.5 and 30°C Sorghum bicolor
530
-
 sinapyl aldehyde wild type isoform CAD2, at pH 6.5 and 30°C Sorghum bicolor
920
-
 coniferyl aldehyde isoform CAD4 mutant L119W/G301F, at pH 6.5 and 30°C Sorghum bicolor