ID   3HBH1_PSEAC             Reviewed;         394 AA.
AC   Q9F131;
DT   01-SEP-2009, integrated into UniProtKB/Swiss-Prot.
DT   01-MAR-2001, sequence version 1.
DT   03-AUG-2022, entry version 60.
DE   RecName: Full=3-hydroxybenzoate 6-hydroxylase 1;
DE            EC=1.14.13.24;
DE   AltName: Full=Constitutive 3-hydroxybenzoate 6-hydroxylase;
GN   Name=xlnD;
OS   Pseudomonas alcaligenes.
OC   Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC   Pseudomonadaceae; Pseudomonas.
OX   NCBI_TaxID=43263;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND INDUCTION.
RC   STRAIN=NCIMB 9867 / P25X;
RX   PubMed=12909360; DOI=10.1016/s0378-1119(03)00619-x;
RA   Yeo C.C., Wong M.V.-M., Feng Y., Song K.P., Poh C.L.;
RT   "Molecular characterization of an inducible gentisate 1,2-dioxygenase gene,
RT   xlnE, from Pseudomonas alcaligenes NCIMB 9867.";
RL   Gene 312:239-248(2003).
RN   [2]
RP   CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES,
RP   SUBUNIT, COFACTOR, AND ACTIVITY REGULATION.
RC   STRAIN=NCIMB 9867 / P25X;
RX   PubMed=16267294; DOI=10.1128/jb.187.22.7696-7702.2005;
RA   Gao X., Tan C.L., Yeo C.C., Poh C.L.;
RT   "Molecular and biochemical characterization of the xlnD-encoded 3-
RT   hydroxybenzoate 6-hydroxylase involved in the degradation of 2,5-xylenol
RT   via the gentisate pathway in Pseudomonas alcaligenes NCIMB 9867.";
RL   J. Bacteriol. 187:7696-7702(2005).
CC   -!- FUNCTION: Catalyzes the NAD- or NADP-dependent conversion of 3-
CC       hydroxybenzoate to gentisate. The affinity of the enzyme toward NAD is
CC       twice as high as for NADP. The enzyme shows higher specific activities
CC       against the intermediates in the degradation of 2,5-xylenol and 3,5-
CC       xylenol, 3-hydroxy-4-methylbenzoate and 3-hydroxy-5-methylbenzoate,
CC       respectively, than for 3-hydroxybenzoate. It also shows activity
CC       against 3-substituted benzoates.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=3-hydroxybenzoate + H(+) + NADH + O2 = 2,5-dihydroxybenzoate +
CC         H2O + NAD(+); Xref=Rhea:RHEA:22692, ChEBI:CHEBI:15377,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:16193,
CC         ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:58044;
CC         EC=1.14.13.24; Evidence={ECO:0000269|PubMed:16267294};
CC   -!- COFACTOR:
CC       Name=FAD; Xref=ChEBI:CHEBI:57692;
CC         Evidence={ECO:0000269|PubMed:16267294};
CC   -!- ACTIVITY REGULATION: Inhibited by manganese, copper, mercury, and iron
CC       ions. {ECO:0000269|PubMed:16267294}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=79 uM for 3-hydroxybenzoate (with NAD as cosubstrate)
CC         {ECO:0000269|PubMed:16267294};
CC         KM=108 uM for 3-hydroxybenzoate (with NADP as cosubstrate)
CC         {ECO:0000269|PubMed:16267294};
CC         KM=95 uM for 3-hydroxy-4-methylbenzoate (with NAD as cosubstrate)
CC         {ECO:0000269|PubMed:16267294};
CC         KM=71 uM for 3-hydroxy-4-methylbenzoate (with NADP as cosubstrate)
CC         {ECO:0000269|PubMed:16267294};
CC         KM=112 uM for NADH {ECO:0000269|PubMed:16267294};
CC         KM=225 uM for NADPH {ECO:0000269|PubMed:16267294};
CC       pH dependence:
CC         Optimum pH is 7.5. {ECO:0000269|PubMed:16267294};
CC       Temperature dependence:
CC         Optimum temperature is 25 degrees Celsius.
CC         {ECO:0000269|PubMed:16267294};
CC   -!- SUBUNIT: Homotrimer. {ECO:0000269|PubMed:16267294}.
CC   -!- INDUCTION: Constitutively expressed. Up-regulated by aromatic
CC       substrates, the best one being 3-hydroxybenzoate.
CC       {ECO:0000269|PubMed:12909360}.
CC   -!- SIMILARITY: Belongs to the 3-hydroxybenzoate 6-hydroxylase family.
CC       {ECO:0000305}.
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DR   EMBL; AF173167; AAG39455.1; -; Genomic_DNA.
DR   AlphaFoldDB; Q9F131; -.
DR   SMR; Q9F131; -.
DR   BioCyc; MetaCyc:MON-14775; -.
DR   BRENDA; 1.14.13.24; 5088.
DR   SABIO-RK; Q9F131; -.
DR   GO; GO:0018669; F:3-hydroxybenzoate 6-monooxygenase activity; IEA:UniProtKB-EC.
DR   GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR   GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR   Gene3D; 3.50.50.60; -; 1.
DR   InterPro; IPR002938; FAD-bd.
DR   InterPro; IPR036188; FAD/NAD-bd_sf.
DR   Pfam; PF01494; FAD_binding_3; 1.
DR   SUPFAM; SSF51905; SSF51905; 1.
PE   1: Evidence at protein level;
KW   Aromatic hydrocarbons catabolism; FAD; Flavoprotein; Monooxygenase; NAD;
KW   NADP; Oxidoreductase.
FT   CHAIN           1..394
FT                   /note="3-hydroxybenzoate 6-hydroxylase 1"
FT                   /id="PRO_0000382700"
SQ   SEQUENCE   394 AA;  43435 MW;  09331B384A367E70 CRC64;
     MHNNILIAGA GIGGLSAALG LARKGMRSIV LEKAPELGEI GAGIQLAPNA YHALDALGIG
     EVARQTGVHV DKLLWMDGMT DKEIASVPLA NRFREFFGNP YAVIHRADFH GLLVEACHKT
     GLVEVRTNAE VVDYENFPDR VEAILHDGSC INGAVLVGAD GLWSNVRQKV IGDGDPRVSG
     HTTYRSVIPA EDMPEELRWN MSTAWAGEGC HMVHYPLKGG KVFNLVLTSN SGASEPEAGV
     PVTTDEVFEK FKTMKRRPTS LIHKGNNWKR WVLCDRDPLP NWVDGRVTLL GDAAHPMMQY
     MAQGASMAIE DAVCLAFELG REMDPVSALK KYNRARFART ARVQTYSRYA SDFIYHAKGG
     AAAMRNELMG GMTPTDFFQW INWLYGKETV EKYK