ID   TFDA_CUPPJ              Reviewed;         287 AA.
AC   P10088; B5A9M3; Q46M53;
DT   01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT   01-JUL-1989, sequence version 1.
DT   02-SEP-2026, entry version 148.
DE   RecName: Full=Alpha-ketoglutarate-dependent 2,4-dichlorophenoxyacetate dioxygenase;
DE            Short=2,4-D dioxygenase;
DE            EC=1.14.11.-;
GN   Name=tfdA; OrderedLocusNames=Reut_D6479;
OS   Cupriavidus pinatubonensis (strain JMP 134 / LMG 1197) (Cupriavidus necator
OS   (strain JMP 134)).
OG   Plasmid pJP4, and Plasmid pPJ4.
OC   Bacteria; Pseudomonadati; Pseudomonadota; Betaproteobacteria;
OC   Burkholderiales; Burkholderiaceae; Cupriavidus.
OX   NCBI_TaxID=264198;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   PLASMID=pJP4;
RX   PubMed=3036764; DOI=10.1128/jb.169.7.2950-2955.1987;
RA   Streber W.R., Timmis K.N., Zenk M.H.;
RT   "Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate
RT   monooxygenase gene tfdA of Alcaligenes eutrophus JMP134.";
RL   J. Bacteriol. 169:2950-2955(1987).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   PLASMID=pJP4;
RX   PubMed=15186344; DOI=10.1111/j.1462-2920.2004.00596.x;
RA   Trefault N., De la Iglesia R., Molina A.M., Manzano M., Ledger T.,
RA   Perez-Pantoja D., Sanchez M.A., Stuardo M., Gonzalez B.;
RT   "Genetic organization of the catabolic plasmid pJP4 from Ralstonia eutropha
RT   JMP134 (pJP4) reveals mechanisms of adaptation to chloroaromatic pollutants
RT   and evolution of specialized chloroaromatic degradation pathways.";
RL   Environ. Microbiol. 6:655-668(2004).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=JMP134 / LMG 1197;
RX   PubMed=20206455; DOI=10.1016/j.syapm.2010.01.001;
RA   Baelum J., Jacobsen C.S., Holben W.E.;
RT   "Comparison of 16S rRNA gene phylogeny and functional tfdA gene
RT   distribution in thirty-one different 2,4-dichlorophenoxyacetic acid and 4-
RT   chloro-2-methylphenoxyacetic acid degraders.";
RL   Syst. Appl. Microbiol. 33:67-70(2010).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=JMP134 / LMG 1197; PLASMID=pPJ4;
RX   PubMed=20339589; DOI=10.1371/journal.pone.0009729;
RA   Lykidis A., Perez-Pantoja D., Ledger T., Mavromatis K., Anderson I.J.,
RA   Ivanova N.N., Hooper S.D., Lapidus A., Lucas S., Gonzalez B.,
RA   Kyrpides N.C.;
RT   "The complete multipartite genome sequence of Cupriavidus necator JMP134, a
RT   versatile pollutant degrader.";
RL   PLoS ONE 5:E9729-E9729(2010).
RN   [5]
RP   MUTAGENESIS OF HIS-9; HIS-114; ASP-116; HIS-168; HIS-201; HIS-214; HIS-217;
RP   HIS-236; HIS-246 AND HIS-263.
RC   PLASMID=pJP4;
RX   PubMed=10777523; DOI=10.1074/jbc.275.17.12400;
RA   Hogan D.A., Smith S.R., Saari E.A., McCracken J., Hausinger R.P.;
RT   "Site-directed mutagenesis of 2,4-dichlorophenoxyacetic acid/alpha-
RT   ketoglutarate dioxygenase. Identification of residues involved in
RT   metallocenter formation and substrate binding.";
RL   J. Biol. Chem. 275:12400-12409(2000).
RN   [6]
RP   HYDROXYLATION AT TRP-113, AND IDENTIFICATION BY MASS SPECTROMETRY.
RC   PLASMID=pJP4;
RX   PubMed=11457355; DOI=10.1021/ja005879x;
RA   Liu A., Ho R.Y.N., Que L. Jr., Ryle M.J., Phinney B.S., Hausinger R.P.;
RT   "Alternative reactivity of an alpha-ketoglutarate-dependent iron(II)
RT   oxygenase: enzyme self-hydroxylation.";
RL   J. Am. Chem. Soc. 123:5126-5127(2001).
RN   [7]
RP   3D-STRUCTURE MODELING.
RC   PLASMID=pJP4;
RX   PubMed=11955067; DOI=10.1021/bi016014e;
RA   Elkins J.M., Ryle M.J., Clifton I.J., Dunning Hotopp J.C., Lloyd J.S.,
RA   Burzlaff N.I., Baldwin J.E., Hausinger R.P., Roach P.L.;
RT   "X-ray crystal structure of Escherichia coli taurine/alpha-ketoglutarate
RT   dioxygenase complexed to ferrous iron and substrates.";
RL   Biochemistry 41:5185-5192(2002).
CC   -!- FUNCTION: Involved in degradation of the herbicide 2,4-
CC       dichlorophenoxyacetic acid (2,4-D). Is also able to degrade 2-methyl-4-
CC       chlorophenoxyacetic acid and 3-chlorobenzoic acid.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(2,4-dichlorophenoxy)acetate + 2-oxoglutarate + O2 = 2,4-
CC         dichlorophenol + glyoxylate + succinate + CO2; Xref=Rhea:RHEA:48984,
CC         ChEBI:CHEBI:15379, ChEBI:CHEBI:16526, ChEBI:CHEBI:16738,
CC         ChEBI:CHEBI:16810, ChEBI:CHEBI:19351, ChEBI:CHEBI:30031,
CC         ChEBI:CHEBI:36655;
CC   -!- COFACTOR:
CC       Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC       Note=Binds 1 Fe(2+) ion per subunit.;
CC   -!- ACTIVITY REGULATION: Activated by ascorbate. {ECO:0000305}.
CC   -!- PATHWAY: Xenobiotic degradation; (2,4-dichlorophenoxy)acetate
CC       degradation.
CC   -!- PTM: Hydroxylated on Trp-113; inactivates the enzyme.
CC       {ECO:0000269|PubMed:11457355}.
CC   -!- SIMILARITY: Belongs to the TfdA dioxygenase family. {ECO:0000305}.
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DR   EMBL; M16730; AAA21983.1; -; Genomic_DNA.
DR   EMBL; AY365053; AAR31052.1; -; Genomic_DNA.
DR   EMBL; EU827467; ACF35494.1; -; Genomic_DNA.
DR   EMBL; CP000093; AAZ65777.1; -; Genomic_DNA.
DR   PIR; A27082; A27082.
DR   RefSeq; WP_011178399.1; NZ_AY365053.1.
DR   AlphaFoldDB; P10088; -.
DR   SMR; P10088; -.
DR   GeneID; 55536829; -.
DR   KEGG; reu:Reut_D6479; -.
DR   HOGENOM; CLU_036005_2_0_4; -.
DR   OrthoDB; 581608at2; -.
DR   BioCyc; MetaCyc:MONOMER-14384; -.
DR   UniPathway; UPA00685; -.
DR   GO; GO:0018602; F:2,4-dichlorophenoxyacetate alpha-ketoglutarate dioxygenase activity; IEA:RHEA.
DR   GO; GO:0046300; P:2,4-dichlorophenoxyacetic acid catabolic process; IEA:UniProtKB-UniPathway.
DR   Gene3D; 3.60.130.10; Clavaminate synthase-like; 1.
DR   InterPro; IPR042098; GlaH-like_sf.
DR   InterPro; IPR003819; TauD/TfdA-like.
DR   InterPro; IPR051178; TfdA_dioxygenase.
DR   PANTHER; PTHR43779:SF3; (3R)-3-[(CARBOXYMETHYL)AMINO]FATTY ACID OXYGENASE_DECARBOXYLASE; 1.
DR   PANTHER; PTHR43779; DIOXYGENASE RV0097-RELATED; 1.
DR   Pfam; PF02668; TauD; 1.
DR   SUPFAM; SSF51197; Clavaminate synthase-like; 1.
PE   1: Evidence at protein level;
KW   Aromatic hydrocarbons catabolism; Dioxygenase; Hydroxylation; Iron;
KW   Metal-binding; Oxidoreductase; Plasmid; Vitamin C.
FT   CHAIN           1..287
FT                   /note="Alpha-ketoglutarate-dependent 2,4-
FT                   dichlorophenoxyacetate dioxygenase"
FT                   /id="PRO_0000194017"
FT   BINDING         114
FT                   /ligand="Fe cation"
FT                   /ligand_id="ChEBI:CHEBI:24875"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000305"
FT   BINDING         116
FT                   /ligand="Fe cation"
FT                   /ligand_id="ChEBI:CHEBI:24875"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000305"
FT   BINDING         141
FT                   /ligand="2-oxoglutarate"
FT                   /ligand_id="ChEBI:CHEBI:16810"
FT                   /evidence="ECO:0000250"
FT   BINDING         248
FT                   /ligand="2-oxoglutarate"
FT                   /ligand_id="ChEBI:CHEBI:16810"
FT                   /evidence="ECO:0000250"
FT   BINDING         263
FT                   /ligand="Fe cation"
FT                   /ligand_id="ChEBI:CHEBI:24875"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000305"
FT   BINDING         274
FT                   /ligand="2-oxoglutarate"
FT                   /ligand_id="ChEBI:CHEBI:16810"
FT                   /evidence="ECO:0000250"
FT   BINDING         278
FT                   /ligand="2-oxoglutarate"
FT                   /ligand_id="ChEBI:CHEBI:16810"
FT                   /evidence="ECO:0000250"
FT   MOD_RES         113
FT                   /note="3-hydroxytryptophan; by autocatalysis"
FT                   /evidence="ECO:0000269|PubMed:11457355"
FT   MUTAGEN         9
FT                   /note="H->A: No change in activity and substrate affinity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         114
FT                   /note="H->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         116
FT                   /note="D->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         168
FT                   /note="H->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         201
FT                   /note="H->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         214
FT                   /note="H->A: 10-fold decrease in affinity for 2,4-D and
FT                   highly reduced activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         217
FT                   /note="H->A: 2.5-fold decrease in affinity for 2,4-D but no
FT                   change in activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         236
FT                   /note="H->A: No change in activity and substrate affinity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         246
FT                   /note="H->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
FT   MUTAGEN         263
FT                   /note="H->A: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:10777523"
SQ   SEQUENCE   287 AA;  32316 MW;  0875BA8BE9119E70 CRC64;
     MSVVANPLHP LFAAGVEDID LREALGSTEV REIERLMDEK SVLVFRGQPL SQDQQIAFAR
     NFGPLEGGFI KVNQRPSRFK YAELADISNV SLDGKVAQRD AREVVGNFAN QLWHSDSSFQ
     QPAARYSMLS AVVVPPSGGD TEFCDMRAAY DALPRDLQSE LEGLRAEHYA LNSRFLLGDT
     DYSEAQRNAM PPVNWPLVRT HAGSGRKFLF IGAHASHVEG LPVAEGRMLL AELLEHATQR
     EFVYRHRWNV GDLVMWDNRC VLHRGRRYDI SARRELRRAT TLDDAVV
//