Detail Information for IndEnz0016000113
IED ID IndEnz0016000113
Enzyme Type ID tyrosinase000113
Protein Name Grixazone synthase
EC 1.10.3.15
O-aminophenol oxidase
EC 1.10.3.4
Phenoxazinone synthase
PHS
Gene Name griF SGR_4246
Organism Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350)
Taxonomic Lineage cellular organisms Bacteria Terrabacteria group Actinobacteria Actinomycetia (high G+C Gram-positive bacteria) Streptomycetales Streptomycetaceae Streptomyces Streptomyces griseus group Streptomyces griseus subgroup Streptomyces griseus Streptomyces griseus subsp. griseus Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350)
Enzyme Sequence MVHVRKNHLTMTAEEKRRFVHAVLEIKRRGIYDRFVKLHIQINSTDYLDKETGKRLGHVNPGFLPWHRQYLLKFEQALQKVDPRVTLPYWDWTTDHGENSPLWSDTFMGGNGRPGDRRVMTGPFARRNGWKLNISVIPEGPEDPALNGNYTHDDRDYLVRDFGTLTPDLPTPQELEQTLDLTVYDCPPWNHTSGGTPPYESFRNHLEGYTKFAWEPRLGKLHGAAHVWTGGHMMYIGSPNDPVFFLNHCMIDRCWALWQARHPDVPHYLPTVPTQDVPDLNTPLGPWHTKTPADLLDHTRFYTYDQ
Enzyme Length 306
Uniprot Accession Number B1VTI5
Absorption
Active Site
Activity Regulation ACTIVITY REGULATION: Inhibited by 3-amino-4-hydroxybenzensulfonic acid, 4-hydroxy-3-nitrobenzaldehyde, L-tyrosine, P-hydroxybenzaldehyde. Activated by the copper chaperone GriE. {ECO:0000269|PubMed:16282322}.
Binding Site
Calcium Binding
catalytic Activity CATALYTIC ACTIVITY: Reaction=2 3-amino-4-hydroxybenzoate + H(+) + N-acetyl-L-cysteine + 2 O2 = CO2 + grixazone B + 4 H2O; Xref=Rhea:RHEA:41420, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16526, ChEBI:CHEBI:60005, ChEBI:CHEBI:73483, ChEBI:CHEBI:78236; EC=1.10.3.15; Evidence={ECO:0000269|PubMed:16282322}; CATALYTIC ACTIVITY: Reaction=4 2-aminophenol + 3 O2 = 2 2-aminophenoxazin-3-one + 6 H2O; Xref=Rhea:RHEA:40963, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379, ChEBI:CHEBI:17293, ChEBI:CHEBI:18112; EC=1.10.3.4; Evidence={ECO:0000269|PubMed:16282322};
DNA Binding
EC Number 1.10.3.15; 1.10.3.4
Enzyme Function FUNCTION: Involved in the biosynthesis of the parasiticide antibiotic grixazone. Catalyzes the oxidation of 3,4-aminohydroxybenzoate (3,4-AHBOA) to yield the corresponding quinone imine which is then non-enzymatically conjugated with the thiol group of N-acetylcysteine. The resultant compound is oxidized to its quinone imine enzymatically and is then dimerized non-enzymatically with another quinone imine oxidized by GriF to yield grixazone B. 3,4-aminohydroxybenzaldehyde (3,4-AHBAL) can also be used as substrate to yield grixazone A. In the grixazone biosynthetic pathway, it can also function as an o-aminophenol oxidase that catalyzes the formation of the phenoxazinone chromophore from alpha-aminophenol. It can also use 3,4-dihydroxybenzaldehyde, 2-amino-4-methylphenol and 3,4-dihydroxy-L-phenylalanine (L-DOPA) as substrates. In contrast to tyrosinases, it does not display monophenolase activity. {ECO:0000269|PubMed:16282322, ECO:0000303|PubMed:19268377}.
Temperature Dependency BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 55 degrees Celsius. {ECO:0000269|PubMed:16282322};
PH Dependency BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is between 8.5 and 10.5. {ECO:0000269|PubMed:16282322};
Pathway
nucleotide Binding
Features Chain (1); Metal binding (6)
Keywords Antibiotic biosynthesis;Copper;Metal-binding;Oxidoreductase
Interact With
Induction
Subcellular Location
Modified Residue
Post Translational Modification
Signal Peptide
Structure 3D
Cross Reference PDB -
Mapped Pubmed ID -
Motif
Gene Encoded By
Mass 35,583
Kinetics BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=3.5 mM for alpha-aminophenol {ECO:0000269|PubMed:16282322}; KM=0.58 mM for 3,4-AHBAL {ECO:0000269|PubMed:16282322}; KM=0.75 mM for 2-amino-4-methylphenol {ECO:0000269|PubMed:16282322}; KM=0.41 mM for 3,4-dihydroxybenzaldehyde {ECO:0000269|PubMed:16282322}; KM=19 mM for catechol {ECO:0000269|PubMed:16282322}; KM=5.5 mM for L-DOPA {ECO:0000269|PubMed:16282322}; Note=kcat is 20 sec(-1) with alpha-aminophenol as substrate. kcat is 14 sec(-1) with 3,4-AHBAL as substrate. kcat is 18 sec(-1) with 2-amino-4-methylphenol as substrate. kcat is 0.8 sec(-1) with 3,4-dihydroxybenzaldehyde as substrate. kcat is 12 sec(-1) with catechol as substrate. kcat is 0.066 sec(-1) with L-DOPA as substrate. {ECO:0000269|PubMed:16282322};
Metal Binding METAL 39; /note="Copper A"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"; METAL 58; /note="Copper A"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"; METAL 67; /note="Copper A"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"; METAL 222; /note="Copper B"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"; METAL 226; /note="Copper B"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"; METAL 248; /note="Copper B"; /evidence="ECO:0000305|PubMed:16282322, ECO:0000305|PubMed:19268377"
Rhea ID RHEA:41420; RHEA:40963
Cross Reference Brenda