AMNB_COMTE
ID AMNB_COMTE Reviewed; 312 AA.
AC Q6J1Z6; Q38M41;
DT 26-JUN-2013, integrated into UniProtKB/Swiss-Prot.
DT 26-JUN-2013, sequence version 2.
DT 03-AUG-2022, entry version 58.
DE RecName: Full=2-aminophenol 1,6-dioxygenase subunit beta {ECO:0000303|PubMed:15580337};
DE EC=1.13.11.74;
DE AltName: Full=2-amino-5-chlorophenol 1,6-dioxygenase subunit beta {ECO:0000312|EMBL:ABB13577.1};
DE EC=1.13.11.76;
GN Name=cnbCb {ECO:0000312|EMBL:ABB13577.1};
GN Synonyms=amnB {ECO:0000312|EMBL:AAT35226.1};
OS Comamonas testosteroni (Pseudomonas testosteroni).
OG Plasmid pCNB1.
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Comamonadaceae; Comamonas.
OX NCBI_TaxID=285;
RN [1] {ECO:0000305, ECO:0000312|EMBL:AAT35226.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-8, FUNCTION,
RP CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=CNB-1 {ECO:0000269|PubMed:15580337}; PLASMID=pCNB1;
RX PubMed=15580337; DOI=10.1007/s00203-004-0738-5;
RA Wu J.F., Sun C.W., Jiang C.Y., Liu Z.P., Liu S.J.;
RT "A novel 2-aminophenol 1,6-dioxygenase involved in the degradation of p-
RT chloronitrobenzene by Comamonas strain CNB-1: purification, properties,
RT genetic cloning and expression in Escherichia coli.";
RL Arch. Microbiol. 183:1-8(2005).
RN [2] {ECO:0000305, ECO:0000312|EMBL:ABB13577.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND PATHWAY.
RC STRAIN=CNB-1 {ECO:0000269|PubMed:16517619}; PLASMID=pCNB1;
RX PubMed=16517619; DOI=10.1128/aem.72.3.1759-1765.2006;
RA Wu J.F., Jiang C.Y., Wang B.J., Ma Y.F., Liu Z.P., Liu S.J.;
RT "Novel partial reductive pathway for 4-chloronitrobenzene and nitrobenzene
RT degradation in Comamonas sp. strain CNB-1.";
RL Appl. Environ. Microbiol. 72:1759-1765(2006).
RN [3] {ECO:0000305, ECO:0000312|EMBL:ABB13577.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC STRAIN=CNB-1 {ECO:0000269|PubMed:17526790}; PLASMID=pCNB1;
RX PubMed=17526790; DOI=10.1128/aem.00616-07;
RA Ma Y.F., Wu J.F., Wang S.Y., Jiang C.Y., Zhang Y., Qi S.W., Liu L.,
RA Zhao G.P., Liu S.J.;
RT "Nucleotide sequence of plasmid pCNB1 from Comamonas strain CNB-1 reveals
RT novel genetic organization and evolution for 4-chloronitrobenzene
RT degradation.";
RL Appl. Environ. Microbiol. 73:4477-4483(2007).
RN [4] {ECO:0000305, ECO:0000312|PDB:3VSG}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH ALPHA SUBUNIT AND IN
RP COMPLEXES WITH REACTION INTERMEDIATE; PRODUCT; INHIBITOR AND IRON,
RP COFACTOR, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT,
RP REACTION MECHANISM, AND MUTAGENESIS OF HIS-13; HIS-62; TYR-129; HIS-195 AND
RP GLU-251.
RC STRAIN=CNB-1 {ECO:0000269|PubMed:23275161}; PLASMID=pCNB1;
RX PubMed=23275161; DOI=10.1107/s0907444912042072;
RA Li de F., Zhang J.Y., Hou Y.J., Liu L., Hu Y., Liu S.J., Wang da C.,
RA Liu W.;
RT "Structures of aminophenol dioxygenase in complex with intermediate,
RT product and inhibitor.";
RL Acta Crystallogr. D 69:32-43(2013).
CC -!- FUNCTION: Component of the 2-aminophenol 1,6-dioxygenase (APD) complex
CC that catalyzes the ring fission of 2-aminophenol to produce 2-
CC aminomuconic semialdehyde. CnbCb seems to be the catalytic subunit of
CC the complex. Also active on other substrates such as 2-amino-5-
CC chlorophenol (68% activity), protocatechuate (33% activity) and
CC catechol (5% activity). Both 2-aminophenol and 2-amino-5-cholorophenol
CC are likely native substrates for this dioxygenase which is involved in
CC the reductive degradation pathway of both nitrobenzene (NB) and 4-
CC chloronitrobenzene (4-CNB), allowing C.testosteroni strain CNB-1 to
CC grow on these compounds as sole source of carbon, nitrogen, and energy.
CC {ECO:0000269|PubMed:15580337, ECO:0000269|PubMed:16517619}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-aminophenol + O2 = 2-aminomuconate 6-semialdehyde;
CC Xref=Rhea:RHEA:26305, ChEBI:CHEBI:15379, ChEBI:CHEBI:18112,
CC ChEBI:CHEBI:77634; EC=1.13.11.74;
CC Evidence={ECO:0000269|PubMed:15580337};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-amino-5-chlorophenol + O2 = 2-amino-5-chloromuconate 6-
CC semialdehyde; Xref=Rhea:RHEA:37543, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:75051, ChEBI:CHEBI:75057; EC=1.13.11.76;
CC Evidence={ECO:0000269|PubMed:15580337};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:15580337, ECO:0000269|PubMed:23275161};
CC Note=Binds 2 Fe(2+) ions per APD complex. The iron ions are bound to
CC the beta subunit. {ECO:0000269|PubMed:15580337,
CC ECO:0000269|PubMed:23275161};
CC -!- ACTIVITY REGULATION: Complete loss of activity in the presence of
CC Ni(2+), Co(2+), Cd(2+), Zn(2+) and hydrogen peroxide, however activity
CC with hydrogen peroxide partially restored upon addition of excess
CC ascorbate. Partially inhibited by Fe(2+), Mg(2+), Ca(2+), Mn(2+),
CC Cu(2+) and also by EDTA, at 2 mM concentration. Total activity
CC inhibited in the presence of catechol or 4-nitrocatechol but completely
CC restored after removal of catechol and addition of 2 mM Fe(2+) and 5 mM
CC ascorbate. {ECO:0000269|PubMed:15580337, ECO:0000269|PubMed:23275161}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.89 uM for 2-aminophenol {ECO:0000269|PubMed:15580337};
CC KM=17.1 uM for 2-aminophenol {ECO:0000269|PubMed:23275161};
CC KM=0.77 uM for 2-amino-5-chlorophenol {ECO:0000269|PubMed:15580337};
CC KM=6.84 uM for catechol {ECO:0000269|PubMed:23275161};
CC KM=77.5 uM for oxygen (in the presence of 2-aminophenol)
CC {ECO:0000269|PubMed:23275161};
CC KM=58.3 uM for oxygen (in the presence of catechol)
CC {ECO:0000269|PubMed:23275161};
CC Vmax=44.6 umol/min/mg enzyme with 2-aminophenol as substrate
CC {ECO:0000269|PubMed:15580337};
CC Vmax=19.0 umol/min/mg enzyme with 2-aminophenol as substrate
CC {ECO:0000269|PubMed:23275161};
CC Vmax=19.3 umol/min/mg enzyme with 2-amino-5-chlorophenol as substrate
CC {ECO:0000269|PubMed:15580337};
CC Vmax=1.12 umol/min/mg enzyme with catechol as substrate
CC {ECO:0000269|PubMed:23275161};
CC -!- PATHWAY: Xenobiotic degradation; nitrobenzene degradation.
CC {ECO:0000269|PubMed:16517619, ECO:0000269|PubMed:17526790}.
CC -!- PATHWAY: Xenobiotic degradation; 4-chloronitrobenzene degradation.
CC -!- SUBUNIT: The APD complex is a heterotetramer of 2 alpha (CnbCa) and 2
CC beta (CnbCb) subunits. {ECO:0000269|PubMed:15580337,
CC ECO:0000269|PubMed:23275161}.
CC -!- MISCELLANEOUS: Not active on 4-methylcatechol, 4-chlorocatechol, 2,4-
CC dihydroxybenzoate, o-nitrophenol, p-nitrophenol or 4-nitrocatechol.
CC -!- SIMILARITY: Belongs to the LigB/MhpB extradiol dioxygenase family.
CC {ECO:0000255}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAT35226.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; AY605054; AAT35226.1; ALT_INIT; Genomic_DNA.
DR EMBL; EF079106; ABB13577.1; -; Genomic_DNA.
DR RefSeq; YP_001967698.1; NC_010935.1.
DR PDB; 3VSG; X-ray; 2.40 A; B/D=1-312.
DR PDB; 3VSH; X-ray; 2.70 A; B/D=1-312.
DR PDB; 3VSI; X-ray; 2.50 A; B/D=1-312.
DR PDB; 3VSJ; X-ray; 2.30 A; B/D=1-312.
DR PDBsum; 3VSG; -.
DR PDBsum; 3VSH; -.
DR PDBsum; 3VSI; -.
DR PDBsum; 3VSJ; -.
DR AlphaFoldDB; Q6J1Z6; -.
DR SMR; Q6J1Z6; -.
DR KEGG; ag:AAT35226; -.
DR PATRIC; fig|688245.4.peg.54; -.
DR BioCyc; MetaCyc:MON-13344; -.
DR BRENDA; 1.13.11.74; 1590.
DR UniPathway; UPA00923; -.
DR UniPathway; UPA01033; -.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0008198; F:ferrous iron binding; IEA:InterPro.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR CDD; cd07372; 2A5CPDO_B; 1.
DR InterPro; IPR034943; 2A5CPDO_B.
DR InterPro; IPR004183; Xdiol_dOase_suB.
DR Pfam; PF02900; LigB; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aromatic hydrocarbons catabolism; Dioxygenase;
KW Direct protein sequencing; Iron; Metal-binding; Oxidoreductase; Plasmid.
FT CHAIN 1..312
FT /note="2-aminophenol 1,6-dioxygenase subunit beta"
FT /id="PRO_0000422781"
FT BINDING 13
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:23275161"
FT BINDING 62
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:23275161"
FT BINDING 251
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:23275161"
FT MUTAGEN 13
FT /note="H->A: Loss of enzyme activity."
FT /evidence="ECO:0000269|PubMed:23275161"
FT MUTAGEN 62
FT /note="H->A: Loss of enzyme activity."
FT /evidence="ECO:0000269|PubMed:23275161"
FT MUTAGEN 129
FT /note="Y->F: Loss of enzyme activity."
FT /evidence="ECO:0000269|PubMed:23275161"
FT MUTAGEN 195
FT /note="H->Q: Loss of enzyme activity."
FT /evidence="ECO:0000269|PubMed:23275161"
FT MUTAGEN 251
FT /note="E->A: Loss of enzyme activity."
FT /evidence="ECO:0000269|PubMed:23275161"
FT CONFLICT 8
FT /note="G -> E (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000269|PubMed:15580337"
FT STRAND 2..10
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 16..19
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 35..51
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 54..72
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 75..82
FT /evidence="ECO:0007829|PDB:3VSJ"
FT TURN 87..89
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 91..98
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 100..112
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 117..120
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 129..138
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 146..153
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 154..157
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 160..181
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 184..189
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 208..210
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 216..231
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 234..248
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 251..254
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 256..263
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 272..279
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 282..290
FT /evidence="ECO:0007829|PDB:3VSJ"
FT HELIX 291..294
FT /evidence="ECO:0007829|PDB:3VSJ"
FT STRAND 298..301
FT /evidence="ECO:0007829|PDB:3VSJ"
SQ SEQUENCE 312 AA; 35040 MW; E05323C35EB8EFA4 CRC64;
MQGEIIAGFL APHPPHLVYG ENPPQNEPRS QGGWEVLRWA YERARERLDA MKPDVLLVHS
PHWITSVGHH FLGVPELSGK SVDPIFPNVF RYDFSLNVDV ELAEACAEEG RKAGLVTKMM
RNPKFRVDYG TITTLHLIRP QWDIPVVGIS ANNSPYYLNT KEGMSEMDVL GKATREAIRK
TGRKAVLLAS NTLSHWHFHE EPTIPEDMSK EYPATMAGYQ WDIRMIELMR QGKTSEVFKL
LPQFIDEAFA EVKSGAFTWM HAAMQYPELA AELFGYGTVI GTGNAVMEWD LRKAGLSMLG
AADQKQRSAA VA
