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Literature summary extracted from

  • Geueke, B.; Hummel, W.
    A new bacterial L-amino acid oxidase with a broad substrate specificity: purification and characterization (2002), Enzyme Microb. Technol., 31, 77-87.
No PubMed abstract available

General Stability

EC Number General Stability Organism
1.4.3.2 freezing at -18°C without addition of glycerol inactivates the enzyme Rhodococcus opacus
1.4.3.2 stability of the enzyme can be increased by storage or incubation of the enzyme in glycine/NaOH buffer Rhodococcus opacus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.4.3.2 glycine activity strongly decreased in glycine/NaOH-buffer, competitive inhibition, no inhibition, when 10 mM L-phenylanalnine is used as substrate Rhodococcus opacus
1.4.3.2 L-arginine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-citrulline pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-leucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-lysine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-methionine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-norleucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-ornithine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-phenylalanine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-serine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 L-tyrosine pH 7.6, 30°C Rhodococcus opacus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.2 0.015
-
L-lysine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.019
-
L-tyrosine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.022
-
L-phenylalanine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.026
-
L-norleucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.026
-
L-citrulline pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.028
-
L-leucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.028
-
L-asparagine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.034
-
L-ornithine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.039
-
L-methionine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.07
-
L-arginine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.085
-
L-glutamine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.274
-
L-alanine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 0.411
-
L-Glutamic acid pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 1.36
-
L-serine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 3.73
-
L-valine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 5.11
-
L-isoleucine pH 7.6, 30°C Rhodococcus opacus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.2 53000
-
2 * 53000, SDS-PAGE, 2 * 54000, MALDI-TOF/MS, correlates with a homodimer containing 2 FAD molecules Rhodococcus opacus
1.4.3.2 54000
-
2 * 53000, SDS-PAGE, 2 * 54000, MALDI-TOF/MS, correlates with a homodimer containing 2 FAD molecules Rhodococcus opacus
1.4.3.2 99000
-
gel filtration Rhodococcus opacus
1.4.3.2 104000
-
MALDI-TOF/MS Rhodococcus opacus

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.2 Rhodococcus opacus Q8VPD4 DSM 43250, one of the best L-AAO producer
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.2 to homogeneity, recovery of 68% and 144fold Rhodococcus opacus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.3.2 4.6
-
homogenous enzyme, L-alanine as susbtrate, 30°C, pH 7.6 Rhodococcus opacus

Storage Stability

EC Number Storage Stability Organism
1.4.3.2 -18°C, 50 mM glycine/NaOH buffer, pH 8.6, 50% glycerol, stable for several months, withou glycerol inactivation Rhodococcus opacus
1.4.3.2 4°C, 50 mM glycine/NaOH buffer, pH 8.6, stable for several months Rhodococcus opacus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.2 DL-3-(1-pyridyl)alanine + H2O + O2 24% of the activity compared to L-alanine Rhodococcus opacus 3-(1-pyridyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 DL-4-fluorophenylalanine + H2O + O2 25% of the activity compared to L-alanine Rhodococcus opacus (4-fluorophenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 DL-homophenylalanine + H2O + O2 50% of the activity compared to L-alanine Rhodococcus opacus 2-oxo-4-phenylbutanoic acid + NH3 + H2O2
-
?
1.4.3.2 DL-norleucine + H2O + O2 73% of the activity compared to L-alanine Rhodococcus opacus alpha-ketocaproate + NH3 + H2O2
-
?
1.4.3.2 DL-O-methylserine + H2O + O2 99% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 DL-phenylseine + H2O + O2 8% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 L-2,3-diaminopropionic acid + H2O + O2 71% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 L-2,4-diaminobutyric acid + H2O + O2 100% of the activity compared to L-alanine Rhodococcus opacus 4-amino-2-oxobutanoate + NH3 + H2O2
-
?
1.4.3.2 L-2-amino-4,4-dimethylvaleric acid + H2O + O2 46% of the activity compared to L-alanine Rhodococcus opacus 4,4-dimethyl-2-oxopentanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-2-amino-5,5-dimethylhexanoic acid + H2O + O2 44% of the activity compared to L-alanine Rhodococcus opacus 5,5-dimethyl-2-oxohexanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-2-chlorophenylalanine + H2O + O2 24% of the activity compared to L-alanine Rhodococcus opacus (2-chlorophenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-3-(1-naphthyl)alanine + H2O + O2 109% of the activity compared to L-alanine Rhodococcus opacus 3-(1-naphthyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-4-aminophenylalanine + H2O + O2 16% of the activity compared to L-alanine Rhodococcus opacus (4-aminophenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-4-chlorophenylalanine + H2O + O2 9% of the activity compared to L-alanine Rhodococcus opacus (4-chlorophenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-4-nitrophenylalanine + H2O + O2 30% of the activity compared to L-alanine Rhodococcus opacus (4-nitrophenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-alanine + H2O + O2 highly enantioselective enzyme with a broad substrate specificity Rhodococcus opacus 2-oxopropanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-arginine + H2O + O2 70% of the activity compared to L-alanine Rhodococcus opacus 2-oxo-5-guanidinovaleric acid + NH3 + H2O2
-
?
1.4.3.2 L-asparagine + H2O + O2 97% of the activity compared to L-alanine Rhodococcus opacus alpha-ketosuccinamic acid + NH3 + H2O2
-
?
1.4.3.2 L-aspartic acid + H2O + O2 11% of the activity compared to L-alanine Rhodococcus opacus oxaloacetate + NH3 + H2O2
-
?
1.4.3.2 L-citrulline + H2O + O2 74% of the activity compared to L-alanine Rhodococcus opacus N-carbamoyl-5-amino-2-oxopentanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-cysteic acid + H2O + O2 34% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 L-cystine + H2O + O2 104% of the activity compared to L-alanine Rhodococcus opacus 2-oxo-3-mercaptopropanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-glutamic acid + H2O + O2 52% of the activity compared to L-alanine Rhodococcus opacus 2-oxoglutarate + NH3 + H2O2
-
?
1.4.3.2 L-glutamine + H2O + O2 52% of the activity compared to L-alanine Rhodococcus opacus 5-amino-2,5-dioxopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-histidine + H2O + O2 36% of the activity compared to L-alanine Rhodococcus opacus 3-(1H-imidazol-4-yl)-2-oxopropionic acid + NH3 + H2O2
-
?
1.4.3.2 L-homocysteine thiolactone + H2O + O2 23% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 L-homoserine lactone + H2O + O2 66% of the activity compared to L-alanine Rhodococcus opacus ? + NH3 + H2O2
-
?
1.4.3.2 L-isoleucine + H2O + O2 45% of the activity compared to L-alanine Rhodococcus opacus alpha-keto-beta-methyl-pentanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-leucine + H2O + O2 68% of the activity compared to L-alanine Rhodococcus opacus 4-methyl-2-oxopentanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-lysine + H2O + O2 55% of the activity compared to L-alanine Rhodococcus opacus 6-amino-2-oxohexanoic acid + NH3 + H2O2
-
?
1.4.3.2 L-methionine + H2O + O2 72% of the activity compared to L-alanine Rhodococcus opacus 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
?
1.4.3.2 L-norleucine + H2O + O2
-
Rhodococcus opacus 2-oxohexanoate + NH3 + H2O2
-
r
1.4.3.2 L-ornithine + H2O + O2 117% of the activity compared to L-alanine Rhodococcus opacus 2-oxo-5-aminopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-phenylalanine + H2O + O2 53% of the activity compared to L-alanine Rhodococcus opacus phenylpyruvate + NH3 + H2O2
-
?
1.4.3.2 L-phenylglycine + H2O + O2 5% of the activity compared to L-alanine Rhodococcus opacus phenylacetate + NH3 + H2O2
-
?
1.4.3.2 L-serine + H2O + O2 72% of the activity compared to L-alanine Rhodococcus opacus 2-oxo-3-hydroxypropionic acid + NH3 + H2O2
-
?
1.4.3.2 L-tryptophan + H2O + O2 12% of the activity compared to L-alanine Rhodococcus opacus indole-3-pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-tyrosine + H2O + O2 54% of the activity compared to L-alanine Rhodococcus opacus p-hydroxyphenylpyruvate + NH3 + H2O2
-
?
1.4.3.2 L-valine + H2O + O2 32% of the activity compared to L-alanine Rhodococcus opacus 2-oxoisovalerate + NH3 + H2O2
-
?
1.4.3.2 Nepsilon-acetyl-L-lysine + H2O + O2 46% of the activity compared to L-alanine Rhodococcus opacus 6-acetylamino-2-oxohexanoic acid + NH3 + H2O2
-
?

Subunits

EC Number Subunits Comment Organism
1.4.3.2 dimer 2 * 53000, SDS-PAGE, 2 * 54000, MALDI-TOF/MS, correlates with a homodimer containing 2 FAD molecules Rhodococcus opacus

Synonyms

EC Number Synonyms Comment Organism
1.4.3.2 L-AAO
-
Rhodococcus opacus
1.4.3.2 L-amino acid:O2 oxidoreductase (deaminating)
-
Rhodococcus opacus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.3.2 25 50 inactivation in 50 mM triethylamine/HCl buffer, pH 7.6 in the same range as 50 mM potassium-phosphate buffer pH 6.6 Rhodococcus opacus
1.4.3.2 30
-
incubation for 30 min with 0.28 M H2O2 results in 50% inactivation Rhodococcus opacus
1.4.3.2 37
-
t1/2: 35 min, increases more than 20fold by incubating the L-AAO in glycine/NaOH buffer: 938 min, potassium phosphate buffer t1/2: 44 min Rhodococcus opacus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.2 8 9 for 10 mM L-alanine, L-leucine, L-phenylalanine, the pH-profiles are almost identical Rhodococcus opacus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.4.3.2 4 11 L-phenylalanine as substrate, pH 4: ca. 10% of the activity, pH 11: ca. 60% of the activity Rhodococcus opacus
1.4.3.2 4.5 10.5 L-alanine as substrate, pH 4.5, ca. 3% of the activity, pH 10.5, ca. 30% of the activity Rhodococcus opacus

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.2 FAD a homodimer complex containing 2 FAD molecules Rhodococcus opacus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.4.3.2 1.06
-
L-tyrosine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 2.28
-
L-phenylalanine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 2.91
-
L-norleucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 4.5
-
glycine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 5.42
-
L-arginine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 5.69
-
L-leucine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 6.84
-
L-methionine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 12.5
-
L-ornithine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 20.6
-
L-citrulline pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 63.7
-
L-lysine pH 7.6, 30°C Rhodococcus opacus
1.4.3.2 192
-
L-serine pH 7.6, 30°C Rhodococcus opacus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.4.3.2 Rhodococcus opacus isoelectric focusing, using a narrow pH-range of 4-6
-
4.8