Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.24.27 extracted from

  • Kusano, M.; Yasukawa, K.; Inouye, K.
    Synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester catalyzed by thermolysin variants with improved activity (2010), Enzyme Microb. Technol., 46, 320-325.
No PubMed abstract available

Application

Application Comment Organism
synthesis thermolysin is industrially used for the synthesis of N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester, a precursor of an artificial sweetener aspartame, from N-carbobenzoxy-L-aspartic acid and L-phenylalanine methyl ester Bacillus thermoproteolyticus

Protein Variants

Protein Variants Comment Organism
D150A mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type Bacillus thermoproteolyticus
D150E mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type Bacillus thermoproteolyticus
D150W mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type Bacillus thermoproteolyticus
F114A mutant with reduced activity Bacillus thermoproteolyticus
I168A mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type Bacillus thermoproteolyticus
N227H mutant with improved activity, the mutant has higher kcat values in N-carbobenzoxy-L-Asp-L-Phe-methyl ester synthesis than wild type Bacillus thermoproteolyticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.7
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
0.8
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1.3
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.1
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.1
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
5.8
-
L-phenylalanine methyl ester mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
7
-
L-phenylalanine methyl ester mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
7.3
-
L-phenylalanine methyl ester mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
8.7
-
L-phenylalanine methyl ester wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
8.8
-
L-phenylalanine methyl ester mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
11.3
-
L-phenylalanine methyl ester mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
12.9
-
L-phenylalanine methyl ester mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
49
-
N-carbobenzoxy-L-aspartic acid mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
54
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
57
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
60
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
62
-
N-carbobenzoxy-L-aspartic acid wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
70
-
N-carbobenzoxy-L-aspartic acid mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
105
-
N-carbobenzoxy-L-aspartic acid mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus

Organism

Organism UniProt Comment Textmining
Bacillus thermoproteolyticus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-carbobenzoxy-L-Asp-L-Phe-methyl ester + H2O
-
Bacillus thermoproteolyticus N-carbobenzoxy-L-aspartic acid + L-phenylalanine methyl ester
-
r
N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O
-
Bacillus thermoproteolyticus ?
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
L-phenylalanine methyl ester mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
0.3
-
N-carbobenzoxy-L-aspartic acid mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1.3
-
L-phenylalanine methyl ester wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1.3
-
N-carbobenzoxy-L-aspartic acid wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
1.5
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.1
-
L-phenylalanine methyl ester mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.1
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.3
-
L-phenylalanine methyl ester mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.3
-
N-carbobenzoxy-L-aspartic acid mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.5
-
L-phenylalanine methyl ester mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
2.5
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
3.2
-
L-phenylalanine methyl ester mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
3.2
-
N-carbobenzoxy-L-aspartic acid mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
4.9
-
L-phenylalanine methyl ester mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
4.9
-
N-carbobenzoxy-L-aspartic acid mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
9.5
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
12.3
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
17.1
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
20.5
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus
24.4
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
49.4
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
around pH 7.0 Bacillus thermoproteolyticus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme F114A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
9.3
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester wild type enzyme, at pH 7.5, at 25°C Bacillus thermoproteolyticus
11.7
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
13.7
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme N227H, at pH 7.5, at 25°C Bacillus thermoproteolyticus
16
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme I168A, at pH 7.5, at 25°C Bacillus thermoproteolyticus
23
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150W, at pH 7.5, at 25°C Bacillus thermoproteolyticus
27.6
-
N-carbobenzoxy-L-Asp-L-Phe-methyl ester mutant enzyme D150E, at pH 7.5, at 25°C Bacillus thermoproteolyticus