IED Industry Enzyme Database

Detail Information for IndEnz0016000039
IED ID IndEnz0016000039
Enzyme Type ID tyrosinase000039
Protein Name Laccase
EC 1.10.3.2
Bilirubin oxidase
EC 1.3.3.5
Spore coat protein A
Gene Name cotA pig BSU06300
Organism Bacillus subtilis (strain 168)
Taxonomic Lineage cellular organisms Bacteria Terrabacteria group Firmicutes Bacilli Bacillales Bacillaceae Bacillus Bacillus subtilis group Bacillus subtilis Bacillus subtilis subsp. subtilis Bacillus subtilis (strain 168)
Enzyme Sequence MTLEKFVDALPIPDTLKPVQQSKEKTYYEVTMEECTHQLHRDLPPTRLWGYNGLFPGPTIEVKRNENVYVKWMNNLPSTHFLPIDHTIHHSDSQHEEPEVKTVVHLHGGVTPDDSDGYPEAWFSKDFEQTGPYFKREVYHYPNQQRGAILWYHDHAMALTRLNVYAGLVGAYIIHDPKEKRLKLPSDEYDVPLLITDRTINEDGSLFYPSAPENPSPSLPNPSIVPAFCGETILVNGKVWPYLEVEPRKYRFRVINASNTRTYNLSLDNGGDFIQIGSDGGLLPRSVKLNSFSLAPAERYDIIIDFTAYEGESIILANSAGCGGDVNPETDANIMQFRVTKPLAQKDESRKPKYLASYPSVQHERIQNIRTLKLAGTQDEYGRPVLLLNNKRWHDPVTETPKVGTTEIWSIINPTRGTHPIHLHLVSFRVLDRRPFDIARYQESGELSYTGPAVPPPPSEKGWKDTIQAHAGEVLRIAATFGPYSGRYVWHCHILEHEDYDMMRPMDITDPHK
Enzyme Length 513
Uniprot Accession Number P07788
Absorption
Active Site
Activity Regulation ACTIVITY REGULATION: Inhibited by azide. {ECO:0000269|PubMed:20200715}.
Binding Site
Calcium Binding
catalytic Activity CATALYTIC ACTIVITY: Reaction=4 hydroquinone + O2 = 4 benzosemiquinone + 2 H2O; Xref=Rhea:RHEA:11276, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379, ChEBI:CHEBI:17594, ChEBI:CHEBI:17977; EC=1.10.3.2; Evidence={ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:20200715, ECO:0000269|PubMed:22481612, ECO:0000269|PubMed:27050268}; CATALYTIC ACTIVITY: Reaction=2 bilirubin IXalpha + O2 = 2 biliverdin IXalpha + 2 H2O; Xref=Rhea:RHEA:20980, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379, ChEBI:CHEBI:57977, ChEBI:CHEBI:57991; EC=1.3.3.5; Evidence={ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226};
DNA Binding
EC Number 1.10.3.2; 1.3.3.5
Enzyme Function FUNCTION: Multicopper oxidase that catalyzes the oxidation of a variety of substrates, including phenolic and non-phenolic compounds. Substrates include syringaldazine (SGZ), 2,6-dimethoxyphenol (2,6-DMP) and the non-phenolic compound 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) (PubMed:11514528, PubMed:11884407, PubMed:18307408, PubMed:20200715, PubMed:22481612, PubMed:27050268). Has no tyrosinase activity (PubMed:11514528). Is implicated in the biosynthesis of a brownish pigment that characterizes sporulating colonies of B.subtilis, and which appears to be a melanin-like product and to confer protection against UV light (PubMed:2821284, PubMed:11514528, PubMed:11884407). {ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:20200715, ECO:0000269|PubMed:22481612, ECO:0000269|PubMed:27050268, ECO:0000269|PubMed:2821284}.; FUNCTION: In vitro, also shows strong bilirubin oxidase (BOD) activity, and can catalyze the oxidation of free bilirubin (UB), direct bilirubin (conjugated with glucuronic acid, DB) and ditaurobilirubin. {ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226}.
Temperature Dependency BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Highly thermostable (PubMed:11884407, PubMed:16391148). Optimum temperature is 45 degrees Celsius for SGZ oxidation (PubMed:11514528). Optimum temperature is 75 degrees Celsius with ABTS as substrate (PubMed:11884407). {ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:16391148};
PH Dependency BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 7 for SGZ and bilirubin oxidation (PubMed:11514528, PubMed:11884407, PubMed:16391148). Optimum pH is about 4 for ditaurobilirubin oxidation (PubMed:16391148). Optimum pH is below 3.0 for ABTS oxidation (PubMed:11884407). Optimum pH is 8.0 for UB oxidation, whereas optimum pH for DB oxidation is 5.5 (PubMed:33618226). {ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226};
Pathway
nucleotide Binding
Features Beta strand (36); Chain (1); Domain (4); Erroneous initiation (2); Helix (11); Metal binding (12); Mutagenesis (22); Sequence conflict (3); Site (2); Turn (3)
Keywords 3D-structure;Copper;Metal-binding;Oxidoreductase;Reference proteome;Repeat;Sporulation
Interact With
Induction INDUCTION: Expression is switched on about four to five hours after the onset of sporulation, a time that corresponds approximately to the stage of spore coat synthesis and deposition. {ECO:0000269|PubMed:3135411}.
Subcellular Location SUBCELLULAR LOCATION: Spore coat {ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:2821284}. Note=Localized to the surface layers of the endospore. {ECO:0000269|PubMed:11884407}.
Modified Residue
Post Translational Modification
Signal Peptide
Structure 3D X-ray crystallography (24)
Cross Reference PDB 1GSK; 1OF0; 1W6L; 1W6W; 1W8E; 2BHF; 2WSD; 2X87; 2X88; 3ZDW; 4A66; 4A67; 4A68; 4AKO; 4AKP; 4AKQ; 4Q89; 4Q8B; 4YVN; 4YVU; 5ZLJ; 5ZLK; 5ZLL; 5ZLM;
Mapped Pubmed ID -
Motif
Gene Encoded By
Mass 58,499
Kinetics BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=106 uM for ABTS {ECO:0000269|PubMed:11884407}; KM=124 uM for ABTS {ECO:0000269|PubMed:18307408}; KM=120 uM for ABTS {ECO:0000269|PubMed:22481612}; KM=26 uM for SGZ {ECO:0000269|PubMed:11884407}; KM=18 uM for SGZ {ECO:0000269|PubMed:18307408}; KM=227 uM for 2,6-DMP {ECO:0000269|PubMed:18307408}; KM=300 uM for 2,6-DMP {ECO:0000269|PubMed:22481612}; KM=8 uM for bilirubin {ECO:0000269|PubMed:16391148}; KM=15 uM for ditaurobilirubin {ECO:0000269|PubMed:16391148}; KM=162 uM for free bilirubin (under neutral conditions) {ECO:0000269|PubMed:33618226}; KM=45 uM for direct bilirubin (under neutral conditions) {ECO:0000269|PubMed:33618226}; KM=25 uM for O(2) {ECO:0000269|PubMed:20200715}; KM=20 uM for O(2) {ECO:0000269|PubMed:22481612}; Vmax=22 umol/min/mg enzyme with ABTS as substrate {ECO:0000269|PubMed:11884407}; Vmax=4 umol/min/mg enzyme with SGZ as substrate {ECO:0000269|PubMed:11884407}; Vmax=28 umol/min/mg enzyme with bilirubin as substrate {ECO:0000269|PubMed:16391148}; Vmax=10 umol/min/mg enzyme with ditaurobilirubin as substrate {ECO:0000269|PubMed:16391148}; Note=kcat is 16.8 sec(-1) with ABTS as substrate (PubMed:11884407). kcat is 322 sec(-1) with ABTS as substrate (PubMed:18307408). kcat is 144 sec(-1) with ABTS as substrate (PubMed:22481612). kcat is 3.7 sec(-1) with SGZ as substrate (PubMed:11884407). kcat is 80 sec(-1) with SGZ as substrate (PubMed:18307408). kcat is 36 sec(-1) with 2,6-DMP as substrate (PubMed:18307408). kcat is 33 sec(-1) with 2,6-DMP as substrate (PubMed:22481612). kcat is 1.59 sec(-1) with free bilirubin as substrate (PubMed:33618226). kcat is 4.77 sec(-1) with direct bilirubin as substrate (PubMed:33618226). kcat is 140 sec(-1) for O(2) consumption (PubMed:22481612). {ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:22481612, ECO:0000269|PubMed:33618226};
Metal Binding METAL 105; /note="Copper 1; type 2"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW"; METAL 107; /note="Copper 2; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 153; /note="Copper 2; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 155; /note="Copper 3; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 419; /note="Copper 4; type 1"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 422; /note="Copper 1; type 2"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW"; METAL 424; /note="Copper 3; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 491; /note="Copper 3; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 492; /note="Copper 4; type 1"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 493; /note="Copper 2; type 3"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 497; /note="Copper 4; type 1"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"; METAL 502; /note="Copper 4; type 1"; /evidence="ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88, ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"
Rhea ID RHEA:11276; RHEA:20980
Cross Reference Brenda 1.3.3.5;