Literature summary for 1.4.99.B3 extracted from

Motta, P.; Molla, G.; Pollegioni, L.; Nardini, M.
Structure-function relationships in L-amino acid deaminase, a flavoprotein belonging to a novel class of biotechnologically relevant enzymes (2016), J. Biol. Chem., 291, 10457-75.

Search for more literature for 1.4.99.B3

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Proteus myxofaciens

Crystallization (Commentary)

Crystallization (Comment)	Organism
the overall fold resembles that of known amine or amino acid oxidases. An additional alpha+beta subdomain is placed close to the putative transmembrane alpha-helix and to the active-site entrance, an FAD isoalloxazine ring is exposed to solvent, and a large and accessible active site suits to bind large hydrophobic substrates. The enzyme requires substrate-induced conformational changes of part of the active site, particularly in Arg316 and Phe318, to achieve the correct geometry for catalysis	Proteus myxofaciens

Crystallization (Comment)

Organism

the overall fold resembles that of known amine or amino acid oxidases. An additional alpha+beta subdomain is placed close to the putative transmembrane alpha-helix and to the active-site entrance, an FAD isoalloxazine ring is exposed to solvent, and a large and accessible active site suits to bind large hydrophobic substrates. The enzyme requires substrate-induced conformational changes of part of the active site, particularly in Arg316 and Phe318, to achieve the correct geometry for catalysis

Proteus myxofaciens

Protein Variants

Protein Variants	Comment	Organism
additional information	a deleltion mutant lacking the N-terminal transmembrane alpha-helix, residues 1-27, i almost completely soluble, catalytic activity is below detection	Proteus myxofaciens

General Stability

General Stability	Organism
crude extract, 50% residual activity after 24 h at 25°C	Proteus myxofaciens

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	no substrate inhibition up to 100 mM L-phenylalanine	Proteus myxofaciens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
3.27	-	L-phenylalanine	pH 7.5, 25°C	Proteus myxofaciens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	presene of a membrane environment is required for activity	Proteus myxofaciens	16020	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
44600	-	gel filtration, recombinant N-terminal deletion mutant	Proteus myxofaciens
49900	-	-	Proteus myxofaciens

Organism

Organism	UniProt	Comment	Textmining
Proteus myxofaciens	A0A158RFS7	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
2.9	-	pH 7.5, 25°C	Proteus myxofaciens

Storage Stability

Storage Stability	Organism
-20°C, 10% glycerol, 1 month, 50% residual activity	Proteus myxofaciens
-80°C, 10% glycerol, 1 month, 75% residual activity	Proteus myxofaciens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
L-cysteine + H2O + 2 cytochrome b	-	Proteus myxofaciens	2-oxo-3-thiopropanoate + NH3 + 2 reduced cytochrome b	43% of the activitywith L-phenylalanine	?
L-Dopa + H2O + 2 cytochrome b	-	Proteus myxofaciens	3-(3,4-dihydroxyphenyl)-pyruvate + NH3 + 2 reduced cytochrome b	56.7% of the activitywith L-phenylalanine	?
L-isoleucine + H2O + 2 cytochrome b	-	Proteus myxofaciens	3-methyl-2-oxo-pentanoate + NH3 + 2 reduced cytochrome b	43% of the activitywith L-phenylalanine	?
L-leucine + H2O + 2 cytochrome b	-	Proteus myxofaciens	2-oxo-4-methylpentanoate + NH3 + 2 reduced cytochrome b	99.2% of the activitywith L-phenylalanine	?
L-methionine + H2O + 2 cytochrome b	-	Proteus myxofaciens	4-methylsulfanyl-2-oxobutanoate + NH3 + 2 reduced cytochrome b	86% of the activitywith L-phenylalanine	?
L-phenylalanine + H2O + 2 cytochrome b	-	Proteus myxofaciens	phenylpyruvate + NH3 + 2 reduced cytochrome b	-	?
L-phenylalanine + H2O + 2,6-dichlorophenolindophenol	-	Proteus myxofaciens	phenylpyruvate + NH3 + reduced 2,6-dichlorophenolindophenol	membrane-free enzyme can be assayed using 2,6-dichlorophenolindophenol as acceptor	?
L-phenylalanine ethyl ester + H2O + 2 cytochrome b	-	Proteus myxofaciens	phenylpyruvate ethyl ester + NH3 + 2 reduced cytochrome b	33% of the activitywith L-phenylalanine	?
L-tryptophan + H2O + 2 cytochrome b	-	Proteus myxofaciens	3-(1H-indol-3-yl)-2-oxopropanoic acid + NH3 + 2 reduced cytochrome b	60.5% of the activitywith L-phenylalanine	?
L-tyrosine + H2O + 2 cytochrome b	-	Proteus myxofaciens	4-hydroxyphenylpyruvate + NH3 + 2 reduced cytochrome b	13.7% of the activitywith L-phenylalanine	?
additional information	the enzyme does not use dioxygen to reoxidize reduced FADH2 and does not produce hydrogen peroxide, instead, it uses a cytochrome b-like protein as an electron acceptor. No substrates: D-amino acids	Proteus myxofaciens	?	-	?

Subunits

Subunits	Comment	Organism
monomer	1 * 49900, calculated, recombinant N-terminal deletion mutant	Proteus myxofaciens

Synonyms

Synonyms	Comment	Organism
L-amino acid deaminase	-	Proteus myxofaciens
LAAD	-	Proteus myxofaciens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	-	Proteus myxofaciens

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
70	-	negligible activity	Proteus myxofaciens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
8.5	-	L-phenylalanine	pH 7.5, 25°C	Proteus myxofaciens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	7.5	-	Proteus myxofaciens

pH Range

pH Minimum	pH Maximum	Comment	Organism
5	9	negligible activity bleow and above	Proteus myxofaciens

Cofactor

Cofactor	Comment	Organism	Structure
FAD	an FAD isoalloxazine ring is exposed to solvent	Proteus myxofaciens