Literature summary for 1.4.3.2 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 .

Application

Application	Comment	Organism
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)

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

Protein Variants

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

KM Value [mM]

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

Organism

Organism	UniProt	Comment	Textmining
Hebeloma cylindrosporum	S4S6Z0	-	-

Renatured (Commentary)

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
L-alanine + H2O + O2	-	Hebeloma cylindrosporum	2-oxopropanoate + NH3 + H2O2	-	?
L-alanine ethyl ester + H2O + O2	-	Hebeloma cylindrosporum	ethyl 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-canavanine + H2O + O2	-	Hebeloma cylindrosporum	4-[[(diaminomethylidene)amino]oxy]-2-oxobutanoate + 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-oxoglutarate + 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 ethyl ester + H2O + O2	-	Hebeloma cylindrosporum	ethyl 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-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	-	?
L-tyrosine methyl ester + H2O + O2	-	Hebeloma cylindrosporum	methyl 3-(4-hydroxyphenyl)pyruvate + NH3 + H2O2	-	?
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

Synonyms	Comment	Organism
LAAO4	-	Hebeloma cylindrosporum

Turnover Number [1/s]

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

Cofactor

Cofactor	Comment	Organism	Structure
FAD	-	Hebeloma cylindrosporum

kcat/KM [mM/s]

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