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show all sequences of 1.14.14.28

Engineering of LadA for enhanced hexadecane oxidation using random- and site-directed mutagenesis

Dong, Y.; Yan, J.; Du, H.; Chen, M.; Ma, T.; Feng, L.; Appl. Microbiol. Biotechnol. 94, 1019-1029 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Geobacillus thermodenitrificans
Engineering
Amino acid exchange
Commentary
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 [mM]
KM Value Maximum [mM]
Substrate
Commentary
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
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Geobacillus thermodenitrificans
A4IU28
-
-
Geobacillus thermodenitrificans NG80-2
A4IU28
-
-
Purification (Commentary)
Commentary
Organism
wild-type and N-terminal His-tagged fusion proteins
Geobacillus thermodenitrificans
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
docosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-docosanol + FMN + H2O
-
-
-
?
docosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-docosanol + FMN + H2O
-
-
-
?
dotriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-dotriacontanol + FMN + H2O
-
-
-
?
dotriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-dotriacontanol + FMN + H2O
-
-
-
?
hexacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexacosanol + FMN + H2O
-
-
-
?
hexacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexacosanol + FMN + H2O
-
-
-
?
hexadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexadecanol + FMN + H2O
-
-
-
?
hexadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexadecanol + FMN + H2O
-
-
-
?
hexatriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexatriacontanol + FMN + H2O
-
-
-
?
hexatriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexatriacontanol + FMN + H2O
-
-
-
?
octacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-octacosanol + FMN + H2O
-
-
-
?
octadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-octadecanol + FMN + H2O
-
-
-
?
pentadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-pentadecanol + FMN + H2O
-
-
-
?
tetracosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-tetracosanol + FMN + H2O
-
-
-
?
Subunits
Subunits
Commentary
Organism
homodimer
-
Geobacillus thermodenitrificans
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
60
-
wild-type enzyme
Geobacillus thermodenitrificans
65
-
mutant enzyme F146R/N376I
Geobacillus thermodenitrificans
75
-
mutant enzyme F146N/N376I
Geobacillus thermodenitrificans
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
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 [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
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 [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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; 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
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
mutant enzyme F146R/N376I
Geobacillus thermodenitrificans
7.5
-
wild-type enzyme
Geobacillus thermodenitrificans
pH Range
pH Minimum
pH Maximum
Commentary
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
Cofactor
Commentary
Organism
Structure
FMN
-
Geobacillus thermodenitrificans
FMNH2
-
Geobacillus thermodenitrificans
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Geobacillus thermodenitrificans
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FMN
-
Geobacillus thermodenitrificans
FMNH2
-
Geobacillus thermodenitrificans
Engineering (protein specific)
Amino acid exchange
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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
Purification (Commentary) (protein specific)
Commentary
Organism
wild-type and N-terminal His-tagged fusion proteins
Geobacillus thermodenitrificans
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
docosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-docosanol + FMN + H2O
-
-
-
?
docosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-docosanol + FMN + H2O
-
-
-
?
dotriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-dotriacontanol + FMN + H2O
-
-
-
?
dotriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-dotriacontanol + FMN + H2O
-
-
-
?
hexacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexacosanol + FMN + H2O
-
-
-
?
hexacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexacosanol + FMN + H2O
-
-
-
?
hexadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexadecanol + FMN + H2O
-
-
-
?
hexadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexadecanol + FMN + H2O
-
-
-
?
hexatriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-hexatriacontanol + FMN + H2O
-
-
-
?
hexatriacontan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans NG80-2
1-hexatriacontanol + FMN + H2O
-
-
-
?
octacosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-octacosanol + FMN + H2O
-
-
-
?
octadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-octadecanol + FMN + H2O
-
-
-
?
pentadecane + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-pentadecanol + FMN + H2O
-
-
-
?
tetracosan + FMNH2 + O2
-
737528
Geobacillus thermodenitrificans
1-tetracosanol + FMN + H2O
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
-
Geobacillus thermodenitrificans
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
60
-
wild-type enzyme
Geobacillus thermodenitrificans
65
-
mutant enzyme F146R/N376I
Geobacillus thermodenitrificans
75
-
mutant enzyme F146N/N376I
Geobacillus thermodenitrificans
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
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 [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
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 [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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; 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 (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
mutant enzyme F146R/N376I
Geobacillus thermodenitrificans
7.5
-
wild-type enzyme
Geobacillus thermodenitrificans
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
8.8
both the wild-type and the mutants are active at pH values from 6.0 to 8.8
Geobacillus thermodenitrificans
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
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
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
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
Other publictions for EC 1.14.14.28
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
744855
Boonmak
Cloning and expression of thr ...
Geobacillus thermoleovorans, Geobacillus thermoleovorans B23
Extremophiles
18
515-523
2014
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
745580
Jain
Homology modeling and protein ...
Burkholderia thailandensis, Burkholderia thailandensis MSMB121
J. Mol. Model.
20
2340
2014
-
-
1
1
3
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
737528
Dong
Engineering of LadA for enhanc ...
Geobacillus thermodenitrificans, Geobacillus thermodenitrificans NG80-2
Appl. Microbiol. Biotechnol.
94
1019-1029
2012
-
-
1
-
15
-
-
10
-
-
-
-
-
4
-
-
1
-
-
-
-
-
14
1
3
1
1
9
2
1
-
2
-
-
-
-
-
1
2
-
15
-
-
-
-
10
-
-
-
-
-
-
-
1
-
-
-
-
14
1
3
1
1
9
2
1
-
-
-
-
-
-
10
10
738256
Wang
Diversity of flavin-binding mo ...
Geobacillus thermodenitrificans, Geobacillus thermodenitrificans NG80-2
FEMS Microbiol. Ecol.
80
523-533
2012
-
-
-
-
-
-
-
-
-
-
-
2
-
7
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
736632
Li
Crystal structure of long-chai ...
Geobacillus thermodenitrificans, Geobacillus thermodenitrificans NG80-2
J. Mol. Biol.
376
453-465
2008
-
-
1
1
5
-
-
-
-
-
2
2
-
7
-
-
1
-
-
-
-
-
4
1
-
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
5
-
-
-
-
-
-
-
2
2
-
-
-
1
-
-
-
-
4
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
739525
Feng
Genome and proteome of long-ch ...
Geobacillus thermodenitrificans, Geobacillus thermodenitrificans NG80-2
Proc. Natl. Acad. Sci. USA
104
5602-5607
2007
-
2
-
-
-
-
-
-
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-