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Literature summary extracted from

  • Bloess, S.; Beuel, T.; Krueger, T.; Sewald, N.; Dierks, T.; Fischer von Mollard, G.
    Expression, characterization, and site-specific covalent immobilization of an L-amino acid oxidase from the fungus Hebeloma cylindrosporum (2019), Appl. Microbiol. Biotechnol., 103, 2229-2241 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.4.3.2 synthesis purified aldehyde-tagged 6His-hcLAAO4 is covalently bound to a hexylamine resin via the Calpha-formylglycine residue. The immobilized enzyme can be reused repeatedly to generate phenylpyruvate from L-phenylalanine with a total turnover number of 17600 and is stable for over 40 days at 25 °C Hebeloma cylindrosporum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.2 expression of isoform LAAO4 without signal sequence as fusion protein with a 6His tag in Escherichia coli Hebeloma cylindrosporum

Protein Variants

EC Number Protein Variants Comment Organism
1.4.3.2 additional information presence of an N-terminal aldehyde tag does not change the properties of the enzyme Hebeloma cylindrosporum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.2 0.26
-
L-leucine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.26
-
L-phenylalanine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.27
-
L-leucine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.46
-
L-glutamine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.48
-
L-phenylalanine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.56
-
L-glutamine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.67
-
L-leucine methyl ester acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 0.69
-
L-leucine methyl ester untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.2 Hebeloma cylindrosporum S4S6Z0
-
-

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.4.3.2 recombinant purified LAAO4 is activated by exposure to acidic pH, the detergent sodium dodecyl sulfate, or freezing Hebeloma cylindrosporum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.2 L-alanine + H2O + O2
-
Hebeloma cylindrosporum 2-oxopropanoate + NH3 + H2O2
-
?
1.4.3.2 L-alanine ethyl ester + H2O + O2
-
Hebeloma cylindrosporum ethyl 2-oxopropanoate + NH3 + H2O2
-
?
1.4.3.2 L-arginine + H2O + O2
-
Hebeloma cylindrosporum 2-oxo-5-guanidinopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-asparagine + H2O + O2
-
Hebeloma cylindrosporum 2-oxo-3-phenylpropanoate + NH3 + H2O2
-
?
1.4.3.2 L-canavanine + H2O + O2
-
Hebeloma cylindrosporum 4-[[(diaminomethylidene)amino]oxy]-2-oxobutanoate + NH3 + H2O2
-
?
1.4.3.2 L-glutamate + H2O + O2
-
Hebeloma cylindrosporum 2-oxoglutarate + NH3 + H2O2
-
?
1.4.3.2 L-glutamate dimethyl ester + H2O + O2
-
Hebeloma cylindrosporum 2-oxoglutarate dimethyl ester + NH3 + H2O2
-
?
1.4.3.2 L-glutamine + H2O + O2
-
Hebeloma cylindrosporum 2-oxoglutarate + NH3 + H2O2
-
?
1.4.3.2 L-histidine + H2O + O2
-
Hebeloma cylindrosporum 3-(1H-imidazol-4-yl)-2-oxopropanoate + NH3 + H2O2
-
?
1.4.3.2 L-isoleucine + H2O + O2
-
Hebeloma cylindrosporum 3-methyl-2-oxo-pentanoate + NH3 + H2O2
-
?
1.4.3.2 L-leucine + H2O + O2
-
Hebeloma cylindrosporum 4-methyl-2-oxopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-leucine ethyl ester + H2O + O2
-
Hebeloma cylindrosporum ethyl 4-methyl-2-oxopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-leucine methyl ester + H2O + O2
-
Hebeloma cylindrosporum methyl 4-methyl-2-oxopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-lysine + H2O + O2
-
Hebeloma cylindrosporum 6-amino-2-oxohexanoate + NH3 + H2O2
-
?
1.4.3.2 L-methionine + H2O + O2
-
Hebeloma cylindrosporum 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2
-
?
1.4.3.2 L-norleucine + H2O + O2
-
Hebeloma cylindrosporum 2-oxohexanoate + NH3 + H2O2
-
?
1.4.3.2 L-ornithine + H2O + O2
-
Hebeloma cylindrosporum 5-amino-2-oxopentanoate + NH3 + H2O2
-
?
1.4.3.2 L-phenylalanine + H2O + O2
-
Hebeloma cylindrosporum 2-oxo-3-phenylpropanoate + NH3 + H2O2
-
?
1.4.3.2 L-phenylalanine methyl ester + H2O + O2
-
Hebeloma cylindrosporum methyl 2-oxo-3-phenylpropanoate + NH3 + H2O2
-
?
1.4.3.2 L-tyrosine + H2O + O2
-
Hebeloma cylindrosporum 3-(4-hydroxyphenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 L-tyrosine methyl ester + H2O + O2
-
Hebeloma cylindrosporum methyl 3-(4-hydroxyphenyl)pyruvate + NH3 + H2O2
-
?
1.4.3.2 additional information enzyme converts 14 proteinogenic L-amino acids with L-glutamine, L-leucine, L-methionine, L-phenylalanine, L-tyrosine, and L-lysine being the best substrates. Methyl esters of these L-amino acids are also accepted, and ethyl esters are converted but with lower activity. No substrate: L-tert-leucine Hebeloma cylindrosporum ?
-
-

Synonyms

EC Number Synonyms Comment Organism
1.4.3.2 LAAO4
-
Hebeloma cylindrosporum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.3.2 0.83
-
L-phenylalanine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 2.32
-
L-leucine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 2.56
-
L-glutamine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 2.6
-
L-leucine methyl ester untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 21.9
-
L-phenylalanine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 27.8
-
L-leucine methyl ester acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 38.8
-
L-glutamine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 45.3
-
L-leucine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.2 FAD
-
Hebeloma cylindrosporum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4.3.2 3.14
-
L-phenylalanine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 3.77
-
L-leucine methyl ester untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 5.58
-
L-glutamine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 8.48
-
L-leucine untreated recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 41.5
-
L-leucine methyl ester acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 45.2
-
L-phenylalanine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 69.4
-
L-glutamine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum
1.4.3.2 176.2
-
L-leucine acid-activited recombinant protein, pH 7, 30°C Hebeloma cylindrosporum