BRENDA - Enzyme Database

Improving kinetic or thermodynamic stability of an azoreductase by directed evolution

Brissos, V.; Goncalves, N.; Melo, E.; Martins, L.; PLoS ONE 9, e87209 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.6.5.10
recombinant expression in Escherichia coli strains Tuner(DE3) and KRX
Pseudomonas putida
1.7.1.6
recombinant expression in Escherichia coli strains Tuner(DE3) and KRX
Pseudomonas putida
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.6.5.10
additional information
improvement of the kinetic and thermodynamic stability of the azoreductase by directed evolution via rational design approaches, five rounds of mutagenesis/recombination are followed by high-throughput screening. Mutant 1B6 shows a 300fold higher half-life at 50°C compared to the wild-type enzyme. mutant 1B6 has a folded state slightly less stable than the wild-type (with lower melting and optimal temperatures) but in contrast is more resistant to irreversible denaturation. The superior kinetic stability of 1B6 variant is therefore related to an increased resistance of the unfolded monomers to aggregation through the introduction of mutations that disturb hydrophobic patches and increase the surface net charge of the protein. Mutants 2A1 and 2A1-Y179H show increased thermodynamic stability with a 10-20°C higher melting temperature than wild-type, these residues are mostly involved in strengthening the solvent-exposed loops or the inter-dimer interactions of the folded state. Molecular details of mutations that improve stability, overview
Pseudomonas putida
1.6.5.10
Q192R
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant B1G6
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 16B7
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/A48P
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 2A1
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 23C10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A31S/K74E/A88G/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2F11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A77T/F98L/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 3B9
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A88G/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 1B6
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/K74E/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 6F11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A77T/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 14D4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/D7H/A178D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 13G10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/E36D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 1C11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/I6V/T79R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 32F5
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/K74E/A88G
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23C5
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/L161M/L169P
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 27E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/N14D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 6F10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 19E4
Pseudomonas putida
1.6.5.10
Q192R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 12B8
Pseudomonas putida
1.6.5.10
Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant K7E3
Pseudomonas putida
1.7.1.6
additional information
improvement of the kinetic and thermodynamic stability of the azoreductase by directed evolution via rational design approaches, five rounds of mutagenesis/recombination are followed by high-throughput screening. Mutant 1B6 shows a 300fold higher half-life at 50°C compared to the wild-type enzyme. mutant 1B6 has a folded state slightly less stable than the wild-type (with lower melting and optimal temperatures) but in contrast is more resistant to irreversible denaturation. The superior kinetic stability of 1B6 variant is therefore related to an increased resistance of the unfolded monomers to aggregation through the introduction of mutations that disturb hydrophobic patches and increase the surface net charge of the protein. Mutants 2A1 and 2A1-Y179H show increased thermodynamic stability with a 10-20°C higher melting temperature than wild-type, these residues are mostly involved in strengthening the solvent-exposed loops or the inter-dimer interactions of the folded state. Molecular details of mutations that improve stability, overview
Pseudomonas putida
1.7.1.6
Q192R
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant B1G6
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 16B7
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/A48P
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 2A1
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 23C10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A31S/K74E/A88G/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2F11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A77T/F98L/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 3B9
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A88G/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 1B6
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/K74E/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 6F11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A77T/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 14D4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/D7H/A178D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 13G10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/E36D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 1C11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/I6V/T79R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 32F5
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/K74E/A88G
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23C5
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/L161M/L169P
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 27E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/N14D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 6F10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 19E4
Pseudomonas putida
1.7.1.6
Q192R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 12B8
Pseudomonas putida
1.7.1.6
Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant K7E3
Pseudomonas putida
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.6.5.10
additional information
-
additional information
wild-type and mutant kinetics and thermodynamics, overview
Pseudomonas putida
1.7.1.6
additional information
-
additional information
wild-type and mutant kinetics and thermodynamics, overview
Pseudomonas putida
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.6.5.10
Pseudomonas putida
-
-
-
1.6.5.10
Pseudomonas putida MET94
-
-
-
1.7.1.6
Pseudomonas putida
-
-
-
1.7.1.6
Pseudomonas putida MET94
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.5.10
additional information
the bifunctional enzyme also shows activity with azo dyes and NAD(P)H as cofactor, cf. EC 1.7.1.6
743650
Pseudomonas putida
?
-
-
-
-
1.6.5.10
additional information
the bifunctional enzyme also shows activity with azo dyes and NAD(P)H as cofactor, cf. EC 1.7.1.6
743650
Pseudomonas putida MET94
?
-
-
-
-
1.6.5.10
NADPH + H+ + 1,2-naphthoquinone-4-sulfonate
i.e. Lawsone
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,2-naphthoquinone-4-sulfonate
i.e. Lawsone
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,4-benzoquinone
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,4-benzoquinone
-
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 2-hydroxy-1,4-naphthoquinone
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 2-hydroxy-1,4-naphthoquinone
-
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + a quinone
-
743650
Pseudomonas putida
NADP+ + a quinol
-
-
-
?
1.6.5.10
NADPH + H+ + a quinone
-
743650
Pseudomonas putida MET94
NADP+ + a quinol
-
-
-
?
1.6.5.10
NADPH + H+ + anthraquinone-2-sulfonic acid
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + catechol
i.e. 1,2-dihydroxybenzene
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.7.1.6
additional information
the bifunctional enzyme shows activity with quinone substrates, anthraquinone-2-sulfonic acid, 1,4-benzoquinone, 1,2-dihydroxybenzene (catechol), 2-hydroxy-1,4-naphthoquinone (Lawsone), and 1,2-naphthoquinone-4-sulfonate, and NADPH as cofactor, cf. EC 1.6.5.10
743650
Pseudomonas putida
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.6.5.10
homodimer
-
Pseudomonas putida
1.7.1.6
homodimer
-
Pseudomonas putida
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.6.5.10
30
-
assay at
Pseudomonas putida
1.7.1.6
30
-
assay at
Pseudomonas putida
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.6.5.10
7
-
assay at
Pseudomonas putida
1.7.1.6
7
-
assay at
Pseudomonas putida
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.7.1.6
NAD(P)H
-
Pseudomonas putida
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.6.5.10
recombinant expression in Escherichia coli strains Tuner(DE3) and KRX
Pseudomonas putida
1.7.1.6
recombinant expression in Escherichia coli strains Tuner(DE3) and KRX
Pseudomonas putida
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.7.1.6
NAD(P)H
-
Pseudomonas putida
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.6.5.10
additional information
improvement of the kinetic and thermodynamic stability of the azoreductase by directed evolution via rational design approaches, five rounds of mutagenesis/recombination are followed by high-throughput screening. Mutant 1B6 shows a 300fold higher half-life at 50°C compared to the wild-type enzyme. mutant 1B6 has a folded state slightly less stable than the wild-type (with lower melting and optimal temperatures) but in contrast is more resistant to irreversible denaturation. The superior kinetic stability of 1B6 variant is therefore related to an increased resistance of the unfolded monomers to aggregation through the introduction of mutations that disturb hydrophobic patches and increase the surface net charge of the protein. Mutants 2A1 and 2A1-Y179H show increased thermodynamic stability with a 10-20°C higher melting temperature than wild-type, these residues are mostly involved in strengthening the solvent-exposed loops or the inter-dimer interactions of the folded state. Molecular details of mutations that improve stability, overview
Pseudomonas putida
1.6.5.10
Q192R
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant B1G6
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 16B7
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/A48P
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 2A1
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 23C10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A31S/K74E/A88G/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2F11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A77T/F98L/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 3B9
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/A88G/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 1B6
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/K74E/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A178D/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 6F11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/A77T/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 14D4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/D7H/A178D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 13G10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/E36D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 1C11
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/I6V/T79R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 32F5
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/K74E/A88G
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23C5
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/L161M/L169P
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 27E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/N14D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 6F10
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/C129S/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23E4
Pseudomonas putida
1.6.5.10
Q192R/A46P/V159A/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 19E4
Pseudomonas putida
1.6.5.10
Q192R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 12B8
Pseudomonas putida
1.6.5.10
Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant K7E3
Pseudomonas putida
1.7.1.6
additional information
improvement of the kinetic and thermodynamic stability of the azoreductase by directed evolution via rational design approaches, five rounds of mutagenesis/recombination are followed by high-throughput screening. Mutant 1B6 shows a 300fold higher half-life at 50°C compared to the wild-type enzyme. mutant 1B6 has a folded state slightly less stable than the wild-type (with lower melting and optimal temperatures) but in contrast is more resistant to irreversible denaturation. The superior kinetic stability of 1B6 variant is therefore related to an increased resistance of the unfolded monomers to aggregation through the introduction of mutations that disturb hydrophobic patches and increase the surface net charge of the protein. Mutants 2A1 and 2A1-Y179H show increased thermodynamic stability with a 10-20°C higher melting temperature than wild-type, these residues are mostly involved in strengthening the solvent-exposed loops or the inter-dimer interactions of the folded state. Molecular details of mutations that improve stability, overview
Pseudomonas putida
1.7.1.6
Q192R
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant B1G6
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 16B7
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/A48P
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 2A1
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 23C10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A31S/K74E/A88G/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2F11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A77T/F98L/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 3B9
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/A88G/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 1B6
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/K74E/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 2E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A178D/N131D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 85°C, 150 min) relative to parental mutant variant 6F11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/A77T/N131D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 14D4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/D7H/A178D
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 13G10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/E36D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 1C11
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/I6V/T79R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 32F5
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/K74E/A88G
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23C5
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/L161M/L169P
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 27E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/N14D/L143Q
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 6F10
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/C129S/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 80°C, 60 min) relative to parental mutant variant 23E4
Pseudomonas putida
1.7.1.6
Q192R/A46P/V159A/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 19E4
Pseudomonas putida
1.7.1.6
Q192R/Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 12B8
Pseudomonas putida
1.7.1.6
Y179H
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant K7E3
Pseudomonas putida
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.6.5.10
additional information
-
additional information
wild-type and mutant kinetics and thermodynamics, overview
Pseudomonas putida
1.7.1.6
additional information
-
additional information
wild-type and mutant kinetics and thermodynamics, overview
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.5.10
additional information
the bifunctional enzyme also shows activity with azo dyes and NAD(P)H as cofactor, cf. EC 1.7.1.6
743650
Pseudomonas putida
?
-
-
-
-
1.6.5.10
additional information
the bifunctional enzyme also shows activity with azo dyes and NAD(P)H as cofactor, cf. EC 1.7.1.6
743650
Pseudomonas putida MET94
?
-
-
-
-
1.6.5.10
NADPH + H+ + 1,2-naphthoquinone-4-sulfonate
i.e. Lawsone
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,2-naphthoquinone-4-sulfonate
i.e. Lawsone
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,4-benzoquinone
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 1,4-benzoquinone
-
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 2-hydroxy-1,4-naphthoquinone
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + 2-hydroxy-1,4-naphthoquinone
-
743650
Pseudomonas putida MET94
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + a quinone
-
743650
Pseudomonas putida
NADP+ + a quinol
-
-
-
?
1.6.5.10
NADPH + H+ + a quinone
-
743650
Pseudomonas putida MET94
NADP+ + a quinol
-
-
-
?
1.6.5.10
NADPH + H+ + anthraquinone-2-sulfonic acid
-
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.6.5.10
NADPH + H+ + catechol
i.e. 1,2-dihydroxybenzene
743650
Pseudomonas putida
NADP+ + ?
-
-
-
?
1.7.1.6
additional information
the bifunctional enzyme shows activity with quinone substrates, anthraquinone-2-sulfonic acid, 1,4-benzoquinone, 1,2-dihydroxybenzene (catechol), 2-hydroxy-1,4-naphthoquinone (Lawsone), and 1,2-naphthoquinone-4-sulfonate, and NADPH as cofactor, cf. EC 1.6.5.10
743650
Pseudomonas putida
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.6.5.10
homodimer
-
Pseudomonas putida
1.7.1.6
homodimer
-
Pseudomonas putida
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.6.5.10
30
-
assay at
Pseudomonas putida
1.7.1.6
30
-
assay at
Pseudomonas putida
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.6.5.10
7
-
assay at
Pseudomonas putida
1.7.1.6
7
-
assay at
Pseudomonas putida
General Information
EC Number
General Information
Commentary
Organism
1.6.5.10
physiological function
Pseudomonas putida MET94 is a bacteria that degrades a wide range of structurally distinct azo dyes with high efficiency and the azoreductase PpAzoR plays a key role in this process
Pseudomonas putida
1.7.1.6
physiological function
Pseudomonas putida MET94 is a bacteria that degrades a wide range of structurally distinct azo dyes with high efficiency and the azoreductase PpAzoR plays a key role in this process
Pseudomonas putida
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.6.5.10
physiological function
Pseudomonas putida MET94 is a bacteria that degrades a wide range of structurally distinct azo dyes with high efficiency and the azoreductase PpAzoR plays a key role in this process
Pseudomonas putida
1.7.1.6
physiological function
Pseudomonas putida MET94 is a bacteria that degrades a wide range of structurally distinct azo dyes with high efficiency and the azoreductase PpAzoR plays a key role in this process
Pseudomonas putida