Cloned (Comment) | Organism |
---|---|
- |
Geobacillus thermodenitrificans |
Protein Variants | Comment | Organism |
---|---|---|
A102D | hydroxylation activity of purified LadA mutant on hexadecane is 2.1fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.3fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C16 to C28 | Geobacillus thermodenitrificans |
A102D/F146C/L320V/N376I | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
A102D/F146C/N376I | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
A102D/L320V | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
A102E | hydroxylation activity of purified LadA mutant on hexadecane is 2.2fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.2fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein | Geobacillus thermodenitrificans |
F146C | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
F146C/L320V/N376I | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
F146C/N376I | hydroxylation activity of purified LadA mutant on hexadecane is 2.9fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.9fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein | Geobacillus thermodenitrificans |
F146E/N376I | hydroxylation activity of purified LadA mutant on hexadecane is 2.0fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.7fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C15 to C28 | Geobacillus thermodenitrificans |
F146N/N376I | hydroxylation activity of purified LadA mutant on hexadecane is 3.4fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 3.4fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. The mutant enzyme shows a shift in optimum temperature from 60°C (for the wild-type enzyme) to 75°C. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C15 to C28 | Geobacillus thermodenitrificans |
F146Q/N376I | hydroxylation activity of purified LadA mutant on hexadecane is 2.3fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.3fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C14 to C24 | Geobacillus thermodenitrificans |
F146R/N376I | hydroxylation activity of purified LadA mutant on hexadecane is 2.5fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.8fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein. The mutant enzyme is more heat resistant than wild-type protein, with more than half of the initial activity being retained after incubation at 60°C for 12 h, compared to a 60°C incubation of 4 h or less resulting in the loss of half of the initial activity in the wild-type and F146N/N376I mutant. The mutant enzyme shows a shift in optimum temperature from 60°C (for the wild-type enzyme) to 65°C. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C15 to C24 | Geobacillus thermodenitrificans |
L320A | hydroxylation activity of purified LadA mutant on hexadecane is 2.2fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.5fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein. Compared to the wild-type enzyme, the mutant enzyme utilizes a narrower spectrum of n-alkanes, including C15 to C22 | Geobacillus thermodenitrificans |
L320V | hydroxylation activity of purified LadA mutant on hexadecane is 2.5fold higher than that of the wild-type enzyme. Hexadecane degradation rate is 2.4fold higher than that of the wild-type enzyme. A Pseudomonas fluorescens KOB2DELTA1 strain expressing the LadA mutant grows more rapidly with hexadecane than the strain expressing wild-type LadA, confirming the enhanced activity of LadA mutant in vivo. Mutant enzyme with the same size as the wild-type LadA protein | Geobacillus thermodenitrificans |
N376I | mutant enzyme completely loses the catalytic activity | Geobacillus thermodenitrificans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.4 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146E/N376I | Geobacillus thermodenitrificans | |
1.5 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102E | Geobacillus thermodenitrificans | |
1.8 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102D | Geobacillus thermodenitrificans | |
2 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146R/N376I | Geobacillus thermodenitrificans | |
2.7 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146Q/N376I | Geobacillus thermodenitrificans | |
2.8 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146C/N376I | Geobacillus thermodenitrificans | |
7.1 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320A | Geobacillus thermodenitrificans | |
8.9 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146N/N376I | Geobacillus thermodenitrificans | |
9.1 | - |
hexadecane | pH 7.5, 60°C, wild-type enzyme | Geobacillus thermodenitrificans | |
11 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320V | Geobacillus thermodenitrificans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus thermodenitrificans | A4IU28 | - |
- |
Geobacillus thermodenitrificans NG80-2 | A4IU28 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type and N-terminal His-tagged fusion proteins | Geobacillus thermodenitrificans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
docosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-docosanol + FMN + H2O | - |
? | |
docosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans NG80-2 | 1-docosanol + FMN + H2O | - |
? | |
dotriacontan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-dotriacontanol + FMN + H2O | - |
? | |
dotriacontan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans NG80-2 | 1-dotriacontanol + FMN + H2O | - |
? | |
hexacosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-hexacosanol + FMN + H2O | - |
? | |
hexacosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans NG80-2 | 1-hexacosanol + FMN + H2O | - |
? | |
hexadecane + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-hexadecanol + FMN + H2O | - |
? | |
hexadecane + FMNH2 + O2 | - |
Geobacillus thermodenitrificans NG80-2 | 1-hexadecanol + FMN + H2O | - |
? | |
hexatriacontan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-hexatriacontanol + FMN + H2O | - |
? | |
hexatriacontan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans NG80-2 | 1-hexatriacontanol + FMN + H2O | - |
? | |
octacosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-octacosanol + FMN + H2O | - |
? | |
octadecane + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-octadecanol + FMN + H2O | - |
? | |
pentadecane + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-pentadecanol + FMN + H2O | - |
? | |
tetracosan + FMNH2 + O2 | - |
Geobacillus thermodenitrificans | 1-tetracosanol + FMN + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | - |
Geobacillus thermodenitrificans |
Synonyms | Comment | Organism |
---|---|---|
LADA | - |
Geobacillus thermodenitrificans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
wild-type enzyme | Geobacillus thermodenitrificans |
65 | - |
mutant enzyme F146R/N376I | Geobacillus thermodenitrificans |
75 | - |
mutant enzyme F146N/N376I | Geobacillus thermodenitrificans |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 90 | both the wild-type and the mutants are active at temperatures ranging from 40°C to 90°C | Geobacillus thermodenitrificans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
incubation of 4 h or less results in the loss of half of the initial activity in the wild-type and F146N/N376I mutant. Mutant enzyme F146R/N376I retains more than half of the initial activity after incubation for 12 h | Geobacillus thermodenitrificans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
hexadecane | pH 7.5, 60°C, wild-type enzyme | Geobacillus thermodenitrificans | |
0.043 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146E/N376I | Geobacillus thermodenitrificans | |
0.045 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102D | Geobacillus thermodenitrificans | |
0.047 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102E | Geobacillus thermodenitrificans | |
0.047 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320A | Geobacillus thermodenitrificans | |
0.05 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146Q/N376I | Geobacillus thermodenitrificans | |
0.053 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320V | Geobacillus thermodenitrificans | |
0.055 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146R/N376I | Geobacillus thermodenitrificans | |
0.063 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146C/N376I | Geobacillus thermodenitrificans | |
0.073 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146N/N376I | Geobacillus thermodenitrificans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
mutant enzyme F146R/N376I | Geobacillus thermodenitrificans |
7.5 | - |
wild-type enzyme | Geobacillus thermodenitrificans |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 8.8 | both the wild-type and the mutants are active at pH values from 6.0 to 8.8 | Geobacillus thermodenitrificans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | - |
Geobacillus thermodenitrificans | |
FMNH2 | - |
Geobacillus thermodenitrificans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0025 | - |
hexadecane | pH 7.5, 60°C, wild-type enzyme | Geobacillus thermodenitrificans | |
0.0048 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320V | Geobacillus thermodenitrificans | |
0.0067 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme L320A | Geobacillus thermodenitrificans | |
0.0082 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146N/N376I | Geobacillus thermodenitrificans | |
0.0185 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146Q/N376I | Geobacillus thermodenitrificans | |
0.022 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146C/N376I | Geobacillus thermodenitrificans | |
0.0245 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102D | Geobacillus thermodenitrificans | |
0.0295 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146R/N376I | Geobacillus thermodenitrificans | |
0.031 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme F146E/N376I | Geobacillus thermodenitrificans | |
0.0315 | - |
hexadecane | pH 7.5, 60°C, mutant enzyme A102E | Geobacillus thermodenitrificans |