Cloned (Comment) | Organism |
---|---|
gene laao/mog, recombinant expression of His-tagged wild-type enzyme L-amino acid oxidase/monooxygenase in Escherichia coli strain BL21(DE3), selenomethionine-labeled enzyme is expressed in Escherichia coli strain BL21 CodonPlus (DE3)-RIL-X | Pseudomonas sp. AIU 813 |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged wild-type and SeMet-labeled enzymes, sitting-drop vapor diffusion method, mixing of 0.001 ml of protein solution, consisting of 20 mg/mL protein in 20 mM HEPES-NaOH, pH 7.6, and 0.001 ml of reservoir solution, consisting of 8% PEG 4000 and 0.1 M Na-acetate, pH 4.6, Se-Met substituted crystals are obtained using reservoir solution consisting of 16% PEG 3350, 0.02 M citric acid, and 0.08 M bis-Tris propane-HCl, pH 8.8, 20°C, X-ray diffraction structure determination and analysis at 1.90 and 2.20 A resolution respectively | Pseudomonas sp. AIU 813 |
Protein Variants | Comment | Organism |
---|---|---|
C254A | site-directed mutagenesis, the mutant shows unaltered lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254D | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254E | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254F | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254G | site-directed mutagenesis, the mutant shows slightly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254H | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254I | site-directed mutagenesis, the mutant enzyme shows 5times higher specific activity of oxidase activity compared to wild-type, while the lysine 2-monooxygenase activity is completely abolished | Pseudomonas sp. AIU 813 |
C254L | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254L | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254M | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254N | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254P | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254Q | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254R | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254S | site-directed mutagenesis, the mutant shows slightly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254T | site-directed mutagenesis, the mutant shows unaltered lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254V | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254W | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254W | site-directed mutagenesis, the mutant shows moderately reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
C254Y | site-directed mutagenesis, the mutant shows highly reduced lysine 2-monooxygenase activity compared to the wild-type | Pseudomonas sp. AIU 813 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | 4-chloromercuribenzoate inhibits only the oxidase activity of the bifunctional enzyme, while the monooxygenase activity is not affected | Pseudomonas sp. AIU 813 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine + O2 | Pseudomonas sp. AIU 813 | - |
5-aminopentanamide + CO2 + H2O | - |
? | |
additional information | Pseudomonas sp. AIU 813 | bifunctional L-amino acid oxidase, EC 1.4.3.2, of Pseudomonas sp. AIU813, renamed as L-amino acid oxidase/monooxygenase (L-AAO/MOG), exhibits L-lysine 2-monooxygenase as well as oxidase activity | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. AIU 813 | W6JQJ6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and and SeMet-labeled enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, anion exchange chromatography, and gel filtration | Pseudomonas sp. AIU 813 |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-lysine + O2 = 5-aminopentanamide + CO2 + H2O | a slight difference of the binding position of a substrate dictates the activity of this type of enzyme as oxidase or monooxygenase, residue C254 is involved | Pseudomonas sp. AIU 813 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.108 | - |
purified recombinant enzyme, pH 7,0, 30°C, substrate L-arginine | Pseudomonas sp. AIU 813 |
0.146 | - |
purified recombinant enzyme, pH 7,0, 30°C, substrate L-ornithine | Pseudomonas sp. AIU 813 |
0.532 | - |
purified recombinant enzyme, pH 7,0, 30°C, substrate L-lysine | Pseudomonas sp. AIU 813 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-arginine + O2 | - |
Pseudomonas sp. AIU 813 | 4-guanidinobutanamide + CO2 + H2O | - |
? | |
L-lysine + O2 | - |
Pseudomonas sp. AIU 813 | 5-aminopentanamide + CO2 + H2O | - |
? | |
L-lysine + O2 | preferred substrate | Pseudomonas sp. AIU 813 | 5-aminopentanamide + CO2 + H2O | - |
? | |
L-ornithine + O2 | - |
Pseudomonas sp. AIU 813 | 4-aminobutanamide + CO2 + H2O | - |
? | |
additional information | bifunctional L-amino acid oxidase, EC 1.4.3.2, of Pseudomonas sp. AIU813, renamed as L-amino acid oxidase/monooxygenase (L-AAO/MOG), exhibits L-lysine 2-monooxygenase as well as oxidase activity | Pseudomonas sp. AIU 813 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure of L-AAO/MOG, overview. The key residue for the activity conversion of L-AAO/MOG, Cys-254, is located near the aromatic cage (Trp418, Phe473, and Trp516) | Pseudomonas sp. AIU 813 |
Synonyms | Comment | Organism |
---|---|---|
L-AAO/MOG | - |
Pseudomonas sp. AIU 813 |
L-amino acid oxidase/monooxygenase | - |
Pseudomonas sp. AIU 813 |
L-lysine 2-monooxygenase | - |
Pseudomonas sp. AIU 813 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas sp. AIU 813 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Pseudomonas sp. AIU 813 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Pseudomonas sp. AIU 813 |
General Information | Comment | Organism |
---|---|---|
evolution | the bifunctional enzyme L-AAO/MOG belongs to the MAO family of enzymes | Pseudomonas sp. AIU 813 |
additional information | three-dimensional structure of L-AAO/MOG, overview. The key residue for the activity conversion of L-AAO/MOG, Cys254, is located near the aromatic cage (Trp418, Phe473, and Trp516). Cys254 is not directly involved in the substrate binding, but the chemical modification by 4-chloromercuribenzoate or C254I mutation has significant impact on the substrate binding via the side chain of Trp516. A slight difference of the binding position of a substrate dictates the activity of this type of enzyme as oxidase or monooxygenase | Pseudomonas sp. AIU 813 |