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Literature summary for 2.1.1.68 extracted from

  • Wang, W.; Su, S.; Wang, S.; Ye, L.; Yu, H.
    Significantly improved catalytic efficiency of caffeic acid O-methyltransferase towards N-acetylserotonin by strengthening its interactions with the unnatural substrates terminal structure (2019), Enzyme Microb. Technol., 125, 1-5 .
    View publication on PubMed
Search for more literature for 2.1.1.68

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
C296F the mutant shows slight activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296F/Q310L the mutant shows about 4.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296F/Q310L/V314T the mutant shows 9.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296H the mutant shows about 3fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296H/Q310L the mutant shows about 3.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296H/Q310L/V314T the mutant shows about 6.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296Y the mutant shows about 2fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296Y/Q310L/V314T the mutant shows about 8fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
C296Y/V314T the mutant shows about 5.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
Q310L the mutant shows about 2fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
Q310L/V314T the mutant shows about 5.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
V314N the mutant shows slight activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana
V314T the mutant shows about 2.5fold activity improvement in O-methylation of N-acetylserotonin compared to the wild type enzyme Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.26
-
 N-acetylserotonin mutant enzyme C296F/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.36
-
 N-acetylserotonin mutant enzyme Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
0.58
-
 N-acetylserotonin mutant enzyme C296F, at pH 7.8 and 37°C Arabidopsis thaliana
0.58
-
 N-acetylserotonin mutant enzyme V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.74
-
 N-acetylserotonin wild type enzyme, at pH 7.8 and 37°C Arabidopsis thaliana

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9FK25
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.00609
-
 wild type enzyme, at pH 7.8 and 37°C Arabidopsis thaliana
0.00945
-
 mutant enzyme V314N, at pH 7.8 and 37°C Arabidopsis thaliana
0.00982
-
 mutant enzyme C296F, at pH 7.8 and 37°C Arabidopsis thaliana
0.01109
-
 mutant enzyme C296Y, at pH 7.8 and 37°C Arabidopsis thaliana
0.01131
-
 mutant enzyme Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
0.01697
-
 mutant enzyme V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.01985
-
 mutant enzyme C296H, at pH 7.8 and 37°C Arabidopsis thaliana
0.02186
-
 mutant enzyme C296H/Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
0.02701
-
 mutant enzyme C296F/Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
0.03342
-
 mutant enzyme Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.03462
-
 mutant enzyme C296A/V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.03913
-
 mutant enzyme C296H/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.04694
-
 mutant enzyme C296Y/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana
0.05787
-
 mutant enzyme C296F/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + N-acetylserotonin
-
 Arabidopsis thaliana S-adenosyl-L-homocysteine + melatonin
-
 ?

Synonyms

Synonyms Comment Organism
caffeic acid O-methyltransferase
-
 Arabidopsis thaliana
COMT
-
 Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.94
-
 N-acetylserotonin wild type enzyme, at pH 7.8 and 37°C Arabidopsis thaliana
4.76
-
 N-acetylserotonin mutant enzyme Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
5.41
-
 N-acetylserotonin mutant enzyme C296F, at pH 7.8 and 37°C Arabidopsis thaliana
11.32
-
 N-acetylserotonin mutant enzyme V314T, at pH 7.8 and 37°C Arabidopsis thaliana
43.79
-
 N-acetylserotonin mutant enzyme C296F/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3990
-
 N-acetylserotonin wild type enzyme, at pH 7.8 and 37°C Arabidopsis thaliana
9370
-
 N-acetylserotonin mutant enzyme C296F, at pH 7.8 and 37°C Arabidopsis thaliana
13110
-
 N-acetylserotonin mutant enzyme Q310L, at pH 7.8 and 37°C Arabidopsis thaliana
19530
-
 N-acetylserotonin mutant enzyme V314T, at pH 7.8 and 37°C Arabidopsis thaliana
171060
-
 N-acetylserotonin mutant enzyme C296F/Q310L/V314T, at pH 7.8 and 37°C Arabidopsis thaliana