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; |