COTA_BACSU
ID COTA_BACSU Reviewed; 513 AA.
AC P07788; O24818;
DT 01-AUG-1988, integrated into UniProtKB/Swiss-Prot.
DT 30-MAY-2000, sequence version 4.
DT 03-AUG-2022, entry version 144.
DE RecName: Full=Laccase {ECO:0000303|PubMed:11514528};
DE EC=1.10.3.2 {ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:20200715, ECO:0000269|PubMed:22481612, ECO:0000269|PubMed:27050268};
DE AltName: Full=Bilirubin oxidase {ECO:0000303|PubMed:16391148};
DE EC=1.3.3.5 {ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226};
DE AltName: Full=Spore coat protein A {ECO:0000305};
GN Name=cotA {ECO:0000303|PubMed:2821284}; Synonyms=pig;
GN OrderedLocusNames=BSU06300;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=8969499; DOI=10.1099/13500872-142-11-3027;
RA Borriss R., Porwollik S., Schroeter R.;
RT "The 52 degrees-55 degrees segment of the Bacillus subtilis chromosome: a
RT region devoted to purine uptake and metabolism, and containing the genes
RT cotA, gabP and guaA and the pur gene cluster within a 34960 bp nucleotide
RT sequence.";
RL Microbiology 142:3027-3031(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168 / Marburg / ATCC 6051 / DSM 10 / JCM 1465 / NBRC 13719 / NCIMB
RC 3610 / NRRL NRS-744 / VKM B-501;
RX PubMed=9455482; DOI=10.1093/dnares/4.5.335;
RA Kasahara Y., Nakai S., Ogasawara N., Yata K., Sadaie Y.;
RT "Sequence analysis of the groESL-cotA region of the Bacillus subtilis
RT genome, containing the restriction/modification system genes.";
RL DNA Res. 4:335-339(1997).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-37, FUNCTION, SUBCELLULAR LOCATION,
RP AND DISRUPTION PHENOTYPE.
RX PubMed=2821284; DOI=10.1016/0022-2836(87)90506-7;
RA Donovan W., Zheng L., Sandman K., Losick R.;
RT "Genes encoding spore coat polypeptides from Bacillus subtilis.";
RL J. Mol. Biol. 196:1-10(1987).
RN [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-34, AND INDUCTION.
RX PubMed=3135411; DOI=10.1016/0022-2836(88)90536-0;
RA Sandman K., Kroos L., Cutting S.M., Youngman P., Losick R.;
RT "Identification of the promoter for a spore coat protein gene in Bacillus
RT subtilis and studies on the regulation of its induction at a late stage of
RT sporulation.";
RL J. Mol. Biol. 200:461-473(1988).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-10.
RC STRAIN=168;
RA Wray L.V. Jr., Ferson A.E., Fisher S.H.;
RL Submitted (JUL-1995) to the EMBL/GenBank/DDBJ databases.
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND COFACTOR.
RX PubMed=11514528; DOI=10.1128/jb.183.18.5426-5430.2001;
RA Hullo M.F., Moszer I., Danchin A., Martin-Verstraete I.;
RT "CotA of Bacillus subtilis is a copper-dependent laccase.";
RL J. Bacteriol. 183:5426-5430(2001).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, SUBCELLULAR LOCATION, AND MUTAGENESIS OF HIS-497 AND MET-502.
RC STRAIN=168 / MB24;
RX PubMed=11884407; DOI=10.1074/jbc.m200827200;
RA Martins L.O., Soares C.M., Pereira M.M., Teixeira M., Costa T., Jones G.H.,
RA Henriques A.O.;
RT "Molecular and biochemical characterization of a highly stable bacterial
RT laccase that occurs as a structural component of the Bacillus subtilis
RT endospore coat.";
RL J. Biol. Chem. 277:18849-18859(2002).
RN [9]
RP COFACTOR, AND CRYSTALLIZATION.
RX PubMed=12198312; DOI=10.1107/s0907444902011575;
RA Enguita F.J., Matias P.M., Martins L.O., Placido D., Henriques A.O.,
RA Carrondo M.A.;
RT "Spore-coat laccase CotA from Bacillus subtilis: crystallization and
RT preliminary X-ray characterization by the MAD method.";
RL Acta Crystallogr. D 58:1490-1493(2002).
RN [10]
RP FUNCTION AS A BILIRUBIN OXIDASE, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=16391148; DOI=10.1128/aem.72.1.972-975.2006;
RA Sakasegawa S., Ishikawa H., Imamura S., Sakuraba H., Goda S., Ohshima T.;
RT "Bilirubin oxidase activity of Bacillus subtilis CotA.";
RL Appl. Environ. Microbiol. 72:972-975(2006).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, DOMAIN, AND
RP MUTAGENESIS OF ASP-116 AND HIS-497.
RX PubMed=22481612; DOI=10.1039/c2dt12067d;
RA Brissos V., Chen Z., Martins L.O.;
RT "The kinetic role of carboxylate residues in the proximity of the
RT trinuclear centre in the O2 reactivity of CotA-laccase.";
RL Dalton Trans. 41:6247-6255(2012).
RN [12]
RP BIOTECHNOLOGY.
RX PubMed=30030754; DOI=10.1007/s12033-018-0103-6;
RA Zhang Y., Dong W., Lv Z., Liu J., Zhang W., Zhou J., Xin F., Ma J.,
RA Jiang M.;
RT "Surface display of bacterial laccase CotA on Escherichia coli cells and
RT its application in industrial dye decolorization.";
RL Mol. Biotechnol. 60:681-689(2018).
RN [13]
RP FUNCTION AS A BILIRUBIN OXIDASE, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND BIOTECHNOLOGY.
RX PubMed=33618226; DOI=10.1016/j.bbrc.2021.01.094;
RA Zhang C., Zhu L., Zhang J., Wang W., Zeng Y., You S., Qi W., Su R., He Z.;
RT "An effective enzymatic assay for pH selectively measuring direct and total
RT bilirubin concentration by using of CotA.";
RL Biochem. Biophys. Res. Commun. 547:192-197(2021).
RN [14] {ECO:0007744|PDB:1GSK}
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH COPPER, COFACTOR,
RP AND SUBUNIT.
RX PubMed=12637519; DOI=10.1074/jbc.m301251200;
RA Enguita F.J., Martins L.O., Henriques A.O., Carrondo M.A.;
RT "Crystal structure of a bacterial endospore coat component. A laccase with
RT enhanced thermostability properties.";
RL J. Biol. Chem. 278:19416-19425(2003).
RN [15] {ECO:0007744|PDB:1OF0}
RP X-RAY CRYSTALLOGRAPHY (2.45 ANGSTROMS) IN COMPLEX WITH COPPER.
RA Enguita F.J., Marcal D., Grenha R., Martins L.O., Henriques A.O.,
RA Carrondo M.A.;
RT "Structural characterization of a bacterial laccase reaction cycle.";
RL Submitted (APR-2003) to the PDB data bank.
RN [16] {ECO:0007744|PDB:3ZDW}
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) IN COMPLEX WITH COPPER AND ABTS, AND
RP COFACTOR.
RX PubMed=14764581; DOI=10.1074/jbc.m314000200;
RA Enguita F.J., Marcal D., Martins L.O., Grenha R., Henriques A.O.,
RA Lindley P.F., Carrondo M.A.;
RT "Substrate and dioxygen binding to the endospore coat laccase from Bacillus
RT subtilis.";
RL J. Biol. Chem. 279:23472-23476(2004).
RN [17] {ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEXES WITH COPPER; PEROXIDE
RP AND AZIDE, AND COFACTOR.
RX PubMed=16234932; DOI=10.1039/b504806k;
RA Bento I., Martins L.O., Gato Lopes G., Armenia Carrondo M., Lindley P.F.;
RT "Dioxygen reduction by multi-copper oxidases; a structural perspective.";
RL Dalton Trans. 21:3507-3513(2005).
RN [18] {ECO:0007744|PDB:2WSD}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF MUTANT ALA-494 IN COMPLEX WITH
RP COPPER, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF LEU-386 AND ILE-494.
RX PubMed=18307408; DOI=10.1042/bj20080166;
RA Durao P., Chen Z., Silva C.S., Soares C.M., Pereira M.M., Todorovic S.,
RA Hildebrandt P., Bento I., Lindley P.F., Martins L.O.;
RT "Proximal mutations at the type 1 copper site of CotA laccase:
RT spectroscopic, redox, kinetic and structural characterization of I494A and
RT L386A mutants.";
RL Biochem. J. 412:339-346(2008).
RN [19] {ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88}
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEX WITH COPPER, COFACTOR,
RP AND DOMAIN.
RX PubMed=20822511; DOI=10.1186/1472-6807-10-28;
RA Bento I., Silva C.S., Chen Z., Martins L.O., Lindley P.F., Soares C.M.;
RT "Mechanisms underlying dioxygen reduction in laccases. Structural and
RT modelling studies focusing on proton transfer.";
RL BMC Struct. Biol. 10:28-28(2010).
RN [20] {ECO:0007744|PDB:4AKO, ECO:0007744|PDB:4AKP, ECO:0007744|PDB:4AKQ}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF MUTANTS ASP-498; LEU-498 AND
RP THR-498 IN COMPLEX WITH COPPER, FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, DOMAIN, AND MUTAGENESIS
RP OF GLU-498.
RX PubMed=20200715; DOI=10.1039/b922734b;
RA Chen Z., Durao P., Silva C.S., Pereira M.M., Todorovic S., Hildebrandt P.,
RA Bento I., Lindley P.F., Martins L.O.;
RT "The role of Glu498 in the dioxygen reactivity of CotA-laccase from
RT Bacillus subtilis.";
RL Dalton Trans. 39:2875-2882(2010).
RN [21] {ECO:0007744|PDB:4A66, ECO:0007744|PDB:4A67, ECO:0007744|PDB:4A68}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF MUTANTS ALA-116; ASN-116 AND
RP GLU-116 IN COMPLEX WITH COPPER.
RX PubMed=22281748; DOI=10.1107/s0907444911054503;
RA Silva C.S., Damas J.M., Chen Z., Brissos V., Martins L.O., Soares C.M.,
RA Lindley P.F., Bento I.;
RT "The role of Asp116 in the reductive cleavage of dioxygen to water in CotA
RT laccase: assistance during the proton-transfer mechanism.";
RL Acta Crystallogr. D 68:186-193(2012).
RN [22] {ECO:0007744|PDB:4Q89, ECO:0007744|PDB:4Q8B}
RP X-RAY CRYSTALLOGRAPHY (1.91 ANGSTROMS) IN COMPLEX WITH COPPER.
RA Xie T., Liu Z.C., Wang G.G.;
RT "The crystal structure of CotA laccase complexed with sinapic acid.";
RL Submitted (APR-2014) to the PDB data bank.
RN [23] {ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH COPPER AND ABTS,
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF ARG-146;
RP ARG-429; LEU-431; ARG-476; ALA-478 AND THR-480.
RX PubMed=27050268; DOI=10.1107/s2053230x1600426x;
RA Liu Z., Xie T., Zhong Q., Wang G.;
RT "Crystal structure of CotA laccase complexed with 2,2-azinobis-(3-
RT ethylbenzothiazoline-6-sulfonate) at a novel binding site.";
RL Acta Crystallogr. F Struct. Biol. Commun. 72:328-335(2016).
RN [24] {ECO:0007744|PDB:5ZLJ, ECO:0007744|PDB:5ZLK, ECO:0007744|PDB:5ZLL, ECO:0007744|PDB:5ZLM}
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF WILD-TYPE AND MUTANTS CYS-491;
RP ALA-493 AND CYS-493 IN COMPLEX WITH COPPER, AND MUTAGENESIS OF HIS-491 AND
RP HIS-493.
RX PubMed=29722464; DOI=10.1002/cbic.201800236;
RA Xie T., Liu Z., Wang G.;
RT "Structural insight into the allosteric coupling of Cu1 site and trinuclear
RT Cu cluster in CotA laccase.";
RL ChemBioChem 19:1502-1506(2018).
CC -!- FUNCTION: Multicopper oxidase that catalyzes the oxidation of a variety
CC of substrates, including phenolic and non-phenolic compounds.
CC Substrates include syringaldazine (SGZ), 2,6-dimethoxyphenol (2,6-DMP)
CC and the non-phenolic compound 2,2'-azino-bis(3-ethylbenzothiazoline-6-
CC sulfonic acid) (ABTS) (PubMed:11514528, PubMed:11884407,
CC PubMed:18307408, PubMed:20200715, PubMed:22481612, PubMed:27050268).
CC Has no tyrosinase activity (PubMed:11514528). Is implicated in the
CC biosynthesis of a brownish pigment that characterizes sporulating
CC colonies of B.subtilis, and which appears to be a melanin-like product
CC and to confer protection against UV light (PubMed:2821284,
CC PubMed:11514528, PubMed:11884407). {ECO:0000269|PubMed:11514528,
CC ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:18307408,
CC ECO:0000269|PubMed:20200715, ECO:0000269|PubMed:22481612,
CC ECO:0000269|PubMed:27050268, ECO:0000269|PubMed:2821284}.
CC -!- FUNCTION: In vitro, also shows strong bilirubin oxidase (BOD) activity,
CC and can catalyze the oxidation of free bilirubin (UB), direct bilirubin
CC (conjugated with glucuronic acid, DB) and ditaurobilirubin.
CC {ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4 hydroquinone + O2 = 4 benzosemiquinone + 2 H2O;
CC Xref=Rhea:RHEA:11276, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17594, ChEBI:CHEBI:17977; EC=1.10.3.2;
CC Evidence={ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407,
CC ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:20200715,
CC ECO:0000269|PubMed:22481612, ECO:0000269|PubMed:27050268};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 bilirubin IXalpha + O2 = 2 biliverdin IXalpha + 2 H2O;
CC Xref=Rhea:RHEA:20980, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57977, ChEBI:CHEBI:57991; EC=1.3.3.5;
CC Evidence={ECO:0000269|PubMed:16391148, ECO:0000269|PubMed:33618226};
CC -!- COFACTOR:
CC Name=Cu(2+); Xref=ChEBI:CHEBI:29036;
CC Evidence={ECO:0000269|PubMed:11514528, ECO:0000269|PubMed:11884407,
CC ECO:0000269|PubMed:12198312, ECO:0000269|PubMed:12637519,
CC ECO:0000269|PubMed:20200715};
CC Note=Binds 4 copper ions per subunit. The 4 copper centers adopt
CC structures classified as type 1, type 2 and type 3.
CC {ECO:0000269|PubMed:12637519, ECO:0000269|PubMed:14764581,
CC ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20200715,
CC ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268};
CC -!- ACTIVITY REGULATION: Inhibited by azide. {ECO:0000269|PubMed:20200715}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=106 uM for ABTS {ECO:0000269|PubMed:11884407};
CC KM=124 uM for ABTS {ECO:0000269|PubMed:18307408};
CC KM=120 uM for ABTS {ECO:0000269|PubMed:22481612};
CC KM=26 uM for SGZ {ECO:0000269|PubMed:11884407};
CC KM=18 uM for SGZ {ECO:0000269|PubMed:18307408};
CC KM=227 uM for 2,6-DMP {ECO:0000269|PubMed:18307408};
CC KM=300 uM for 2,6-DMP {ECO:0000269|PubMed:22481612};
CC KM=8 uM for bilirubin {ECO:0000269|PubMed:16391148};
CC KM=15 uM for ditaurobilirubin {ECO:0000269|PubMed:16391148};
CC KM=162 uM for free bilirubin (under neutral conditions)
CC {ECO:0000269|PubMed:33618226};
CC KM=45 uM for direct bilirubin (under neutral conditions)
CC {ECO:0000269|PubMed:33618226};
CC KM=25 uM for O(2) {ECO:0000269|PubMed:20200715};
CC KM=20 uM for O(2) {ECO:0000269|PubMed:22481612};
CC Vmax=22 umol/min/mg enzyme with ABTS as substrate
CC {ECO:0000269|PubMed:11884407};
CC Vmax=4 umol/min/mg enzyme with SGZ as substrate
CC {ECO:0000269|PubMed:11884407};
CC Vmax=28 umol/min/mg enzyme with bilirubin as substrate
CC {ECO:0000269|PubMed:16391148};
CC Vmax=10 umol/min/mg enzyme with ditaurobilirubin as substrate
CC {ECO:0000269|PubMed:16391148};
CC Note=kcat is 16.8 sec(-1) with ABTS as substrate (PubMed:11884407).
CC kcat is 322 sec(-1) with ABTS as substrate (PubMed:18307408). kcat is
CC 144 sec(-1) with ABTS as substrate (PubMed:22481612). kcat is 3.7
CC sec(-1) with SGZ as substrate (PubMed:11884407). kcat is 80 sec(-1)
CC with SGZ as substrate (PubMed:18307408). kcat is 36 sec(-1) with 2,6-
CC DMP as substrate (PubMed:18307408). kcat is 33 sec(-1) with 2,6-DMP
CC as substrate (PubMed:22481612). kcat is 1.59 sec(-1) with free
CC bilirubin as substrate (PubMed:33618226). kcat is 4.77 sec(-1) with
CC direct bilirubin as substrate (PubMed:33618226). kcat is 140 sec(-1)
CC for O(2) consumption (PubMed:22481612). {ECO:0000269|PubMed:11884407,
CC ECO:0000269|PubMed:18307408, ECO:0000269|PubMed:22481612,
CC ECO:0000269|PubMed:33618226};
CC pH dependence:
CC Optimum pH is 7 for SGZ and bilirubin oxidation (PubMed:11514528,
CC PubMed:11884407, PubMed:16391148). Optimum pH is about 4 for
CC ditaurobilirubin oxidation (PubMed:16391148). Optimum pH is below 3.0
CC for ABTS oxidation (PubMed:11884407). Optimum pH is 8.0 for UB
CC oxidation, whereas optimum pH for DB oxidation is 5.5
CC (PubMed:33618226). {ECO:0000269|PubMed:11514528,
CC ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:16391148,
CC ECO:0000269|PubMed:33618226};
CC Temperature dependence:
CC Highly thermostable (PubMed:11884407, PubMed:16391148). Optimum
CC temperature is 45 degrees Celsius for SGZ oxidation
CC (PubMed:11514528). Optimum temperature is 75 degrees Celsius with
CC ABTS as substrate (PubMed:11884407). {ECO:0000269|PubMed:11514528,
CC ECO:0000269|PubMed:11884407, ECO:0000269|PubMed:16391148};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:11884407,
CC ECO:0000269|PubMed:12637519}.
CC -!- SUBCELLULAR LOCATION: Spore coat {ECO:0000269|PubMed:11884407,
CC ECO:0000269|PubMed:2821284}. Note=Localized to the surface layers of
CC the endospore. {ECO:0000269|PubMed:11884407}.
CC -!- INDUCTION: Expression is switched on about four to five hours after the
CC onset of sporulation, a time that corresponds approximately to the
CC stage of spore coat synthesis and deposition.
CC {ECO:0000269|PubMed:3135411}.
CC -!- DOMAIN: Glu-498, located within the entrance channel of the trinuclear
CC center, plays a key role in the protonation events that occur at the
CC trinuclear center and in its stabilization, controlling therefore the
CC binding of dioxygen and its further reduction (PubMed:20822511,
CC PubMed:20200715). Asp-116, situated in the exit channel of the
CC trinuclear center, and its hydrogen bond connectivity are highly
CC relevant for the overall reactivity of the laccase but do not appear to
CC play a critical structural role (PubMed:22481612).
CC {ECO:0000269|PubMed:20200715, ECO:0000269|PubMed:20822511,
CC ECO:0000269|PubMed:22481612}.
CC -!- DISRUPTION PHENOTYPE: Disruption of the gene does not prevent spore
CC formation, but mutant is blocked in the appearance of the brown pigment
CC characteristic of colonies of wild-type sporulating cells.
CC {ECO:0000269|PubMed:2821284}.
CC -!- BIOTECHNOLOGY: When expressed on the surface of E.coli cells, CotA was
CC shown to possess improved enzymatic properties, including higher
CC thermal stability and stronger inhibitor tolerance. It has high
CC enzymatic activity against three industrial dye types: an anthraquinone
CC dye, Acid Blue 62, a triphenylmethane dye, Malachite Green, and an azo
CC dye, Methyl Orange. It may help decrease the costs and simplify
CC wastewater treatment, and therefore, surface-displayed CotA shows a
CC great potential for industrial-scale dye decolorization.
CC {ECO:0000269|PubMed:30030754}.
CC -!- BIOTECHNOLOGY: CotA is also a promising candidate for the clinical
CC determination of direct bilirubin and total bilirubin, which are
CC significant diagnostic marker of liver function and cancer.
CC {ECO:0000269|PubMed:33618226}.
CC -!- SIMILARITY: Belongs to the multicopper oxidase family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=BAA22774.1; Type=Erroneous initiation; Evidence={ECO:0000305};
CC Sequence=CAA29165.1; Type=Erroneous initiation; Evidence={ECO:0000305};
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; U51115; AAB62305.1; -; Genomic_DNA.
DR EMBL; AB007638; BAA22774.1; ALT_INIT; Genomic_DNA.
DR EMBL; AL009126; CAB12449.1; -; Genomic_DNA.
DR EMBL; X05678; CAA29165.1; ALT_INIT; Genomic_DNA.
DR EMBL; X07512; CAA30392.1; -; Genomic_DNA.
DR EMBL; U31756; AAC44642.1; -; Genomic_DNA.
DR PIR; F69604; F69604.
DR RefSeq; NP_388511.1; NC_000964.3.
DR RefSeq; WP_003243170.1; NZ_JNCM01000032.1.
DR PDB; 1GSK; X-ray; 1.70 A; A=1-513.
DR PDB; 1OF0; X-ray; 2.45 A; A=1-513.
DR PDB; 1W6L; X-ray; 2.00 A; A=1-513.
DR PDB; 1W6W; X-ray; 2.20 A; A=1-513.
DR PDB; 1W8E; X-ray; 2.20 A; A=1-513.
DR PDB; 2BHF; X-ray; 2.50 A; A=1-513.
DR PDB; 2WSD; X-ray; 1.60 A; A=1-513.
DR PDB; 2X87; X-ray; 2.00 A; A=1-513.
DR PDB; 2X88; X-ray; 1.80 A; A=1-513.
DR PDB; 3ZDW; X-ray; 2.45 A; A=1-513.
DR PDB; 4A66; X-ray; 1.95 A; A=1-513.
DR PDB; 4A67; X-ray; 2.10 A; A=1-513.
DR PDB; 4A68; X-ray; 2.00 A; A=1-513.
DR PDB; 4AKO; X-ray; 1.70 A; A=1-513.
DR PDB; 4AKP; X-ray; 2.00 A; A=1-513.
DR PDB; 4AKQ; X-ray; 2.10 A; A=1-513.
DR PDB; 4Q89; X-ray; 2.31 A; A/B=1-513.
DR PDB; 4Q8B; X-ray; 1.91 A; A/B=1-513.
DR PDB; 4YVN; X-ray; 2.30 A; A=1-513.
DR PDB; 4YVU; X-ray; 2.30 A; A=1-513.
DR PDB; 5ZLJ; X-ray; 1.96 A; A=1-513.
DR PDB; 5ZLK; X-ray; 2.60 A; A=1-513.
DR PDB; 5ZLL; X-ray; 2.60 A; A=1-513.
DR PDB; 5ZLM; X-ray; 1.70 A; A=1-513.
DR PDBsum; 1GSK; -.
DR PDBsum; 1OF0; -.
DR PDBsum; 1W6L; -.
DR PDBsum; 1W6W; -.
DR PDBsum; 1W8E; -.
DR PDBsum; 2BHF; -.
DR PDBsum; 2WSD; -.
DR PDBsum; 2X87; -.
DR PDBsum; 2X88; -.
DR PDBsum; 3ZDW; -.
DR PDBsum; 4A66; -.
DR PDBsum; 4A67; -.
DR PDBsum; 4A68; -.
DR PDBsum; 4AKO; -.
DR PDBsum; 4AKP; -.
DR PDBsum; 4AKQ; -.
DR PDBsum; 4Q89; -.
DR PDBsum; 4Q8B; -.
DR PDBsum; 4YVN; -.
DR PDBsum; 4YVU; -.
DR PDBsum; 5ZLJ; -.
DR PDBsum; 5ZLK; -.
DR PDBsum; 5ZLL; -.
DR PDBsum; 5ZLM; -.
DR AlphaFoldDB; P07788; -.
DR SMR; P07788; -.
DR STRING; 224308.BSU06300; -.
DR PaxDb; P07788; -.
DR EnsemblBacteria; CAB12449; CAB12449; BSU_06300.
DR GeneID; 936023; -.
DR KEGG; bsu:BSU06300; -.
DR PATRIC; fig|224308.179.peg.683; -.
DR eggNOG; COG2132; Bacteria.
DR InParanoid; P07788; -.
DR OMA; PYSGRYV; -.
DR PhylomeDB; P07788; -.
DR BioCyc; BSUB:BSU06300-MON; -.
DR BRENDA; 1.3.3.5; 658.
DR SABIO-RK; P07788; -.
DR EvolutionaryTrace; P07788; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0030288; C:outer membrane-bounded periplasmic space; IBA:GO_Central.
DR GO; GO:0005507; F:copper ion binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IBA:GO_Central.
DR GO; GO:0030435; P:sporulation resulting in formation of a cellular spore; IEA:UniProtKB-KW.
DR Gene3D; 2.60.40.420; -; 3.
DR InterPro; IPR001117; Cu-oxidase.
DR InterPro; IPR011706; Cu-oxidase_C.
DR InterPro; IPR045087; Cu-oxidase_fam.
DR InterPro; IPR011707; Cu-oxidase_N.
DR InterPro; IPR008972; Cupredoxin.
DR PANTHER; PTHR11709; PTHR11709; 1.
DR Pfam; PF00394; Cu-oxidase; 1.
DR Pfam; PF07731; Cu-oxidase_2; 1.
DR Pfam; PF07732; Cu-oxidase_3; 2.
DR SUPFAM; SSF49503; SSF49503; 3.
PE 1: Evidence at protein level;
KW 3D-structure; Copper; Metal-binding; Oxidoreductase; Reference proteome;
KW Repeat; Sporulation.
FT CHAIN 1..513
FT /note="Laccase"
FT /id="PRO_0000079257"
FT DOMAIN 45..81
FT /note="Plastocyanin-like 1"
FT /evidence="ECO:0000255"
FT DOMAIN 101..178
FT /note="Plastocyanin-like 2"
FT /evidence="ECO:0000255"
FT DOMAIN 240..318
FT /note="Plastocyanin-like 3"
FT /evidence="ECO:0000255"
FT DOMAIN 378..509
FT /note="Plastocyanin-like 4"
FT /evidence="ECO:0000255"
FT BINDING 105
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="1"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0007744|PDB:1GSK,
FT ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L,
FT ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E,
FT ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87,
FT ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW"
FT BINDING 107
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 153
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 155
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 419
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 422
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="1"
FT /note="type 2 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0007744|PDB:1GSK,
FT ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L,
FT ECO:0007744|PDB:1W6W, ECO:0007744|PDB:1W8E,
FT ECO:0007744|PDB:2BHF, ECO:0007744|PDB:2X87,
FT ECO:0007744|PDB:2X88, ECO:0007744|PDB:3ZDW"
FT BINDING 424
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 491
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="3"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 492
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 493
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="2"
FT /note="type 3 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 497
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:14764581, ECO:0000269|PubMed:16234932,
FT ECO:0000269|PubMed:20822511, ECO:0000269|PubMed:27050268,
FT ECO:0007744|PDB:1GSK, ECO:0007744|PDB:1OF0,
FT ECO:0007744|PDB:1W6L, ECO:0007744|PDB:1W6W,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:3ZDW, ECO:0007744|PDB:4YVN,
FT ECO:0007744|PDB:4YVU"
FT BINDING 502
FT /ligand="Cu cation"
FT /ligand_id="ChEBI:CHEBI:23378"
FT /ligand_label="4"
FT /note="type 1 copper site"
FT /evidence="ECO:0000269|PubMed:12637519,
FT ECO:0000269|PubMed:16234932, ECO:0000269|PubMed:20822511,
FT ECO:0000269|PubMed:27050268, ECO:0007744|PDB:1GSK,
FT ECO:0007744|PDB:1OF0, ECO:0007744|PDB:1W6L,
FT ECO:0007744|PDB:1W8E, ECO:0007744|PDB:2BHF,
FT ECO:0007744|PDB:2X87, ECO:0007744|PDB:2X88,
FT ECO:0007744|PDB:4YVN, ECO:0007744|PDB:4YVU"
FT SITE 116
FT /note="Plays a crucial role in the protonation steps"
FT /evidence="ECO:0000305|PubMed:22481612"
FT SITE 498
FT /note="Plays a crucial role in the protonation steps"
FT /evidence="ECO:0000305|PubMed:20200715,
FT ECO:0000305|PubMed:20822511, ECO:0000305|PubMed:22481612"
FT MUTAGEN 116
FT /note="D->A: 5-fold decrease in catalytic efficiency with
FT ABTS as substrate. 785-fold decrease in catalytic
FT efficiency with 2,6-DMP as substrate."
FT /evidence="ECO:0000269|PubMed:22481612"
FT MUTAGEN 116
FT /note="D->E: 10-fold decrease in catalytic efficiency with
FT ABTS as substrate. 91-fold decrease in catalytic efficiency
FT with 2,6-DMP as substrate."
FT /evidence="ECO:0000269|PubMed:22481612"
FT MUTAGEN 116
FT /note="D->L: 9-fold decrease in catalytic efficiency with
FT ABTS as substrate. Almost loss of activity with 2,6-DMP as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22481612"
FT MUTAGEN 116
FT /note="D->N: 33-fold decrease in catalytic efficiency with
FT ABTS as substrate. Almost loss of activity with 2,6-DMP as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22481612"
FT MUTAGEN 116
FT /note="D->T: 6-fold decrease in catalytic efficiency with
FT ABTS as substrate. Almost loss of activity with 2,6-DMP as
FT substrate."
FT /evidence="ECO:0000269|PubMed:22481612"
FT MUTAGEN 146
FT /note="R->K: 357-fold decrease in catalytic efficiency with
FT ABTS as substrate. 152-fold decrease in catalytic
FT efficiency with SGZ as substrate."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 386
FT /note="L->A: Slight decrease in catalytic efficiency. Shows
FT minimal changes in the structure of the copper centers."
FT /evidence="ECO:0000269|PubMed:18307408"
FT MUTAGEN 429
FT /note="R->K: 25-fold decrease in catalytic efficiency with
FT ABTS as substrate. 30-fold decrease in catalytic efficiency
FT with SGZ as substrate."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 431
FT /note="L->F: Retains approximately 50% of the wild-type
FT activity with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 476
FT /note="R->K: Retains approximately 20% of the wild-type
FT activity with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 478
FT /note="A->F: Retains approximately 70% of the wild-type
FT activity with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 480
FT /note="T->A: Retains approximately 60% of the wild-type
FT activity with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 480
FT /note="T->F: Retains approximately 30% of the wild-type
FT activity with SGZ but does not affect activity with ABTS."
FT /evidence="ECO:0000269|PubMed:27050268"
FT MUTAGEN 491
FT /note="H->C: Decreases copper content. Strong decrease in
FT catalytic efficiency with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:29722464"
FT MUTAGEN 493
FT /note="H->A: Does not affect copper content. Strong
FT decrease in catalytic efficiency with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:29722464"
FT MUTAGEN 493
FT /note="H->C: Decreases copper content. Strong decrease in
FT catalytic efficiency with both ABTS and SGZ."
FT /evidence="ECO:0000269|PubMed:29722464"
FT MUTAGEN 494
FT /note="I->A: Strong decrease in catalytic efficiency.
FT Significant differences in both the type 1 and type 2
FT copper centers."
FT /evidence="ECO:0000269|PubMed:18307408"
FT MUTAGEN 497
FT /note="H->A: Loss of laccase activity. Mutant fails to
FT develop the dark brown phenotype typical of the wild type
FT strain. Decreases copper content."
FT /evidence="ECO:0000269|PubMed:11884407,
FT ECO:0000269|PubMed:22481612"
FT MUTAGEN 498
FT /note="E->D: 9-fold decrease in catalytic efficiency with
FT ABTS as substrate. 26-fold decrease in catalytic efficiency
FT with 2,6-DMP as substrate."
FT /evidence="ECO:0000269|PubMed:20200715"
FT MUTAGEN 498
FT /note="E->L: Almost loss of laccase activity."
FT /evidence="ECO:0000269|PubMed:20200715"
FT MUTAGEN 498
FT /note="E->T: 165-fold decrease in catalytic efficiency with
FT ABTS as substrate. 400-fold decrease in catalytic
FT efficiency with 2,6-DMP as substrate."
FT /evidence="ECO:0000269|PubMed:20200715"
FT MUTAGEN 502
FT /note="M->L: Loss of laccase activity. Mutant fails to
FT develop the dark brown phenotype typical of the wild type
FT strain."
FT /evidence="ECO:0000269|PubMed:11884407"
FT CONFLICT 347..367
FT /note="DESRKPKYLASYPSVQHERIQ -> TKAESRSTSPHTLRYSMKDT (in
FT Ref. 1; AAB62305)"
FT /evidence="ECO:0000305"
FT CONFLICT 414..420
FT /note="PTRGTHP -> RHAEHIL (in Ref. 1; AAB62305)"
FT /evidence="ECO:0000305"
FT CONFLICT 451..458
FT /note="GPAVPPPP -> VRCPAAA (in Ref. 1; AAB62305)"
FT /evidence="ECO:0000305"
FT STRAND 19..21
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 26..37
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 46..51
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 54..56
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 59..63
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 68..74
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 86..88
FT /evidence="ECO:0007829|PDB:2X88"
FT STRAND 104..107
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 125..127
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 128..130
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 137..141
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 148..154
FT /evidence="ECO:0007829|PDB:2WSD"
FT TURN 157..159
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 160..166
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 169..175
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 177..182
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 187..189
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 190..200
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 210..214
FT /evidence="ECO:0007829|PDB:2X88"
FT STRAND 217..219
FT /evidence="ECO:0007829|PDB:5ZLM"
FT STRAND 231..235
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 238..240
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 242..244
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 247..256
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 263..267
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 273..278
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 281..294
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 299..305
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 307..309
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 313..318
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 322..325
FT /evidence="ECO:0007829|PDB:2WSD"
FT TURN 328..332
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 333..338
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 359..361
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 366..378
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 384..388
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 406..413
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 415..417
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 419..425
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 428..436
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 438..444
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 449..451
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 458..460
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 464..469
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 473..480
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 486..493
FT /evidence="ECO:0007829|PDB:2WSD"
FT HELIX 495..498
FT /evidence="ECO:0007829|PDB:2WSD"
FT TURN 499..501
FT /evidence="ECO:0007829|PDB:2WSD"
FT STRAND 503..509
FT /evidence="ECO:0007829|PDB:2WSD"
SQ SEQUENCE 513 AA; 58499 MW; 836B83B458D75F87 CRC64;
MTLEKFVDAL PIPDTLKPVQ QSKEKTYYEV TMEECTHQLH RDLPPTRLWG YNGLFPGPTI
EVKRNENVYV KWMNNLPSTH FLPIDHTIHH SDSQHEEPEV KTVVHLHGGV TPDDSDGYPE
AWFSKDFEQT GPYFKREVYH YPNQQRGAIL WYHDHAMALT RLNVYAGLVG AYIIHDPKEK
RLKLPSDEYD VPLLITDRTI NEDGSLFYPS APENPSPSLP NPSIVPAFCG ETILVNGKVW
PYLEVEPRKY RFRVINASNT RTYNLSLDNG GDFIQIGSDG GLLPRSVKLN SFSLAPAERY
DIIIDFTAYE GESIILANSA GCGGDVNPET DANIMQFRVT KPLAQKDESR KPKYLASYPS
VQHERIQNIR TLKLAGTQDE YGRPVLLLNN KRWHDPVTET PKVGTTEIWS IINPTRGTHP
IHLHLVSFRV LDRRPFDIAR YQESGELSYT GPAVPPPPSE KGWKDTIQAH AGEVLRIAAT
FGPYSGRYVW HCHILEHEDY DMMRPMDITD PHK
