6HN3M_PSEFL
ID 6HN3M_PSEFL Reviewed; 385 AA.
AC P86491;
DT 18-MAY-2010, integrated into UniProtKB/Swiss-Prot.
DT 18-MAY-2010, sequence version 1.
DT 03-AUG-2022, entry version 28.
DE RecName: Full=6-hydroxynicotinate 3-monooxygenase {ECO:0000303|PubMed:10091591};
DE Short=6-HNA monooxygenase;
DE EC=1.14.13.114 {ECO:0000269|PubMed:10091591};
DE Flags: Precursor;
GN Name=nicC;
OS Pseudomonas fluorescens.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=294;
RN [1] {ECO:0000305}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 323-340, FUNCTION,
RP CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RC STRAIN=TN5 {ECO:0000269|PubMed:10091591};
RX PubMed=10091591; DOI=10.1046/j.1432-1327.1999.00124.x;
RA Nakano H., Wieser M., Hurh B., Kawai T., Yoshida T., Yamane T.,
RA Nagasawa T.;
RT "Purification, characterization and gene cloning of 6-hydroxynicotinate 3-
RT monooxygenase from Pseudomonas fluorescens TN5.";
RL Eur. J. Biochem. 260:120-126(1999).
CC -!- FUNCTION: Flavin-dependent monooxygenase (FMO) that catalyzes the
CC decarboxylative hydroxylation of 6-hydroxynicotinic acid (6-HNA) to
CC 2,5-dihydroxypyridine (2,5-DHP) with concomitant oxidation of NADH, a
CC step in the aerobic nicotinate degradation pathway. Uses NADH in
CC preference to NADPH as an electron donor.
CC {ECO:0000269|PubMed:10091591}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=6-hydroxynicotinate + 2 H(+) + NADH + O2 = 2,5-
CC dihydroxypyridine + CO2 + H2O + NAD(+); Xref=Rhea:RHEA:27333,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16364, ChEBI:CHEBI:16526, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57664, ChEBI:CHEBI:57945; EC=1.14.13.114;
CC Evidence={ECO:0000269|PubMed:10091591};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:10091591};
CC Note=Binds 1 FAD molecule per subunit. {ECO:0000269|PubMed:10091591};
CC -!- ACTIVITY REGULATION: Inhibited competitively by nicotinic acid with a
CC Ki of 0.49 mM. Inhibited by thiol-specific compounds p-
CC chloromercuribenzoate, DTNB, Ag(2)SO(4), HgCl(2), CuCl(2) and N-
CC ethylmaleimide. No inhibition by o-phenanthroline, 8-hydroxyquinoline,
CC EDTA, disodium 4,5-dihydroxy-m-benzenedisulfonate, fluoride, azide,
CC KCl, LiCl, NaCl, BaCl(2), MnCl(2), MgCl(2), PBCl, ZnCl(2), CoCl(2),
CC SnCl(2), FeSO(4), FeCl(3), NiCl(2), CdCl(2), AlCl(3), iodoacetic acid,
CC hydro-xylamine, phenylhydrazine, semicarbazide, cysteamine,
CC alpha,alpha-dipyridyl and urea. {ECO:0000269|PubMed:10091591}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=98 uM for 6-hydroxynicotinic acid (at 30 degrees Celsius and pH
CC 7.0) {ECO:0000269|PubMed:10091591};
CC KM=150 uM for 4-hydroxybenzoic acid (at 30 degrees Celsius and pH
CC 7.0) {ECO:0000269|PubMed:10091591};
CC Vmax=6.67 umol/min/mg enzyme toward 6-hydroxynicotinic acid (at 30
CC degrees Celsius and pH 7.0) {ECO:0000269|PubMed:10091591};
CC pH dependence:
CC Optimum pH is 7.6 with 6-hydroxynicotinic acid as substrate. Stable
CC between pH 6.0 and 9.0. {ECO:0000269|PubMed:10091591};
CC Temperature dependence:
CC Optimum temperature is 35 degrees Celsius with 6-hydroxynicotinic
CC acid as substrate. {ECO:0000269|PubMed:10091591};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:10091591}.
CC -!- SIMILARITY: Belongs to the 6-hydroxynicotinate 3-monooxygenase family.
CC {ECO:0000305}.
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DR AlphaFoldDB; P86491; -.
DR SMR; P86491; -.
DR GO; GO:0043731; F:6-hydroxynicotinate 3-monooxygenase activity; IEA:UniProtKB-EC.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0019439; P:aromatic compound catabolic process; IDA:UniProtKB.
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; Direct protein sequencing; FAD;
KW Flavoprotein; Monooxygenase; NAD; NADP; Oxidoreductase; Signal.
FT SIGNAL 1..20
FT /evidence="ECO:0000255"
FT CHAIN 21..385
FT /note="6-hydroxynicotinate 3-monooxygenase"
FT /evidence="ECO:0000255"
FT /id="PRO_0000394198"
FT ACT_SITE 47
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:A0A0H3LKL4"
FT ACT_SITE 215
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:A0A0H3LKL4"
FT BINDING 15
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 34..35
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 47
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 108
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 130
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 294
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 307..308
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
SQ SEQUENCE 385 AA; 42888 MW; EB01E06986DEB760 CRC64;
MSQSPRIAVV GAGLGGAAAA KLLLQEGFNV RVYEQAPSFS RLGAGIHVGP NVMKILRRIG
IEDALNEQGS HPDYWYSRHW QTGDVLAQIP LGDYAVKEYG ASYLTVHRGD FHALLVEALP
DSVMAYGKFL TKVEDRGNVV VMHFADGTTE EADIVIGPDG VNSRIREELL GPELPKYAGY
LAHRAVFPTP EVKAGMLPFD ACVKWWSDDR HMMTYFVTGK ADELYYVTGV PVEKWDLNDR
WLESSKEEMR EAFSGWHPTV QALIDATVEV TKWSLLERDP LPLWSRGRLV LLGDACHPMK
PHMAQGAAMA IEDGAMLARC LKEVGAHNHE LAFALYEANR AERASKVQRI SHDNTWLRTN
EDPSWCFGYD VFNVPLVEPK VKAAA
