Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | enzyme is activated by incubation for 10 min at pH 3 | Hebeloma cylindrosporum |
Application | Comment | Organism |
---|---|---|
synthesis | expression of isoform LAOO4 in the yeast Pichia pastoris with a 9His-tag and comparison with the 6His-LAAO4 expressed in Escherichia coli. The expression of the enzyme with an ER-signal sequence in Pastoris pastoris results in a glycosylated, secreted protein. The enzymatic activity without activation is higher after expression in Pichia pastoris compared to Escherichia coli. Due to treatment with acidic pH, a striking increase of activity can be detected for both expression systems resulting in similar specific activities after acid activation | Hebeloma cylindrosporum |
Cloned (Comment) | Organism |
---|---|
expression in Pichia pastoris | Hebeloma cylindrosporum |
Protein Variants | Comment | Organism |
---|---|---|
N54A/N164A/N193A/N331A | mutation of the potential glycosylatoon sites, mutant enzyme has the same mobility as the lower band of 9His-LAAO4 treated with PNGaseF | Hebeloma cylindrosporum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
L-leucine | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.07 | - |
L-phenylalanine | acid-activited recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.07 | - |
L-phenylalanine | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.16 | - |
L-methionine | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.29 | - |
L-leucine | acid-activited recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.41 | - |
L-methionine | acid-activated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.51 | - |
L-glutamine | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
0.59 | - |
L-glutamine | acid-activited recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
1.12 | - |
L-phenylalanine methyl ester | acid-activited recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
1.85 | - |
L-phenylalanine methyl ester | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
3.01 | - |
L-leucine methyl ester | untreated recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum | |
3.58 | - |
L-leucine methyl ester | acid-activited recombinant protein, pH 7, 30°C | Hebeloma cylindrosporum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
216000 | - |
gel filtration | Hebeloma cylindrosporum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Hebeloma cylindrosporum | S4S6Z0 | isoform LAAO4 | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | sequence contains four putative N-glycosylation sites predicted at amino acid residues N54, N164, N193, and N331 | Hebeloma cylindrosporum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2 | - |
untreated enzyme, expressed as 6His-LAAO4 in Escherichia coli, pH 7, 30°C | Hebeloma cylindrosporum |
20 | 40 | acid-treated enzyme, expressed as 9His-LAAO4 in Pichia pastoris, as well as 6His-LAAO4 expressed in Escherichia coli, pH 7, 30°C | Hebeloma cylindrosporum |
1020 | - |
untreated enzyme, expressed as 9His-LAAO4 in Pichia pastoris, pH 7, 30°C | Hebeloma cylindrosporum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-chloro-DL-phenylalanine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-3-(4-chlorophenyl)-propanoate + NH3 + H2O2 | - |
? | |
DL-homophenylalanine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-4-phenylbutanoate + NH3 + H2O2 | - |
? | |
L-alanine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxopropanoate + NH3 + H2O2 | - |
? | |
L-arginine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-5-guanidinopentanoate + NH3 + H2O2 | - |
? | |
L-asparagine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-3-phenylpropanoate + NH3 + H2O2 | - |
? | |
L-glutamate + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxoglutarate + NH3 + H2O2 | - |
? | |
L-glutamate dimethyl ester + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxoglutarate dimethyl ester + NH3 + H2O2 | - |
? | |
L-glutamine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-4-carbamoylbutanoic acid + NH3 + H2O2 | - |
? | |
L-histidine + H2O + O2 | - |
Hebeloma cylindrosporum | 3-(1H-imidazol-4-yl)-2-oxopropanoate + NH3 + H2O2 | - |
? | |
L-isoleucine + H2O + O2 | - |
Hebeloma cylindrosporum | 3-methyl-2-oxo-pentanoate + NH3 + H2O2 | - |
? | |
L-leucine + H2O + O2 | - |
Hebeloma cylindrosporum | 4-methyl-2-oxopentanoate + NH3 + H2O2 | - |
? | |
L-leucine methyl ester + H2O + O2 | - |
Hebeloma cylindrosporum | methyl 4-methyl-2-oxopentanoate + NH3 + H2O2 | - |
? | |
L-lysine + H2O + O2 | - |
Hebeloma cylindrosporum | 6-amino-2-oxohexanoate + NH3 + H2O2 | - |
? | |
L-methionine + H2O + O2 | - |
Hebeloma cylindrosporum | 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
L-methionine methyl ester + H2O + O2 | - |
Hebeloma cylindrosporum | methyl 4-methylsulfanyl-2-oxobutanoate + NH3 + H2O2 | - |
? | |
L-norleucine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxohexanoate + NH3 + H2O2 | - |
? | |
L-ornithine + H2O + O2 | - |
Hebeloma cylindrosporum | 5-amino-2-oxopentanoate + NH3 + H2O2 | - |
? | |
L-phenylalanine + H2O + O2 | - |
Hebeloma cylindrosporum | 2-oxo-3-phenylpropanoate + NH3 + H2O2 | - |
? | |
L-phenylalanine methyl ester + H2O + O2 | - |
Hebeloma cylindrosporum | methyl 2-oxo-3-phenylpropanoate + NH3 + H2O2 | - |
? | |
L-tyrosine + H2O + O2 | - |
Hebeloma cylindrosporum | 3-(4-hydroxyphenyl)pyruvate + NH3 + H2O2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
trimer | 3 * 69600, calculated from sequence, 3 * 75000, SDS-PAGE of 9His-LAAO4 expressed in Pichia pastoris | Hebeloma cylindrosporum |
Synonyms | Comment | Organism |
---|---|---|
LAAO4 | - |
Hebeloma cylindrosporum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | broad, acid-treated enzyme | Hebeloma cylindrosporum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | - |
complete loss of activity below | Hebeloma cylindrosporum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | FAD loading of the enzyme expressed in Pichia pastoris is 80%-100% in different expressions | Hebeloma cylindrosporum |