6HN3M_BORBR
ID 6HN3M_BORBR Reviewed; 383 AA.
AC A0A0H3LKL4;
DT 03-JUL-2019, integrated into UniProtKB/Swiss-Prot.
DT 16-SEP-2015, sequence version 1.
DT 03-AUG-2022, entry version 26.
DE RecName: Full=6-hydroxynicotinate 3-monooxygenase {ECO:0000303|PubMed:27218267, ECO:0000303|PubMed:30810301};
DE Short=6-HNA monooxygenase {ECO:0000303|PubMed:27218267, ECO:0000303|PubMed:30810301};
DE EC=1.14.13.114 {ECO:0000269|PubMed:27218267, ECO:0000269|PubMed:30810301};
DE Flags: Precursor;
GN Name=nicC {ECO:0000303|PubMed:30810301};
GN OrderedLocusNames=BB1778 {ECO:0000312|EMBL:CAE32275.1};
OS Bordetella bronchiseptica (strain ATCC BAA-588 / NCTC 13252 / RB50)
OS (Alcaligenes bronchisepticus).
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Alcaligenaceae; Bordetella.
OX NCBI_TaxID=257310;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-588 / NCTC 13252 / RB50;
RX PubMed=12910271; DOI=10.1038/ng1227;
RA Parkhill J., Sebaihia M., Preston A., Murphy L.D., Thomson N.R.,
RA Harris D.E., Holden M.T.G., Churcher C.M., Bentley S.D., Mungall K.L.,
RA Cerdeno-Tarraga A.-M., Temple L., James K.D., Harris B., Quail M.A.,
RA Achtman M., Atkin R., Baker S., Basham D., Bason N., Cherevach I.,
RA Chillingworth T., Collins M., Cronin A., Davis P., Doggett J., Feltwell T.,
RA Goble A., Hamlin N., Hauser H., Holroyd S., Jagels K., Leather S.,
RA Moule S., Norberczak H., O'Neil S., Ormond D., Price C., Rabbinowitsch E.,
RA Rutter S., Sanders M., Saunders D., Seeger K., Sharp S., Simmonds M.,
RA Skelton J., Squares R., Squares S., Stevens K., Unwin L., Whitehead S.,
RA Barrell B.G., Maskell D.J.;
RT "Comparative analysis of the genome sequences of Bordetella pertussis,
RT Bordetella parapertussis and Bordetella bronchiseptica.";
RL Nat. Genet. 35:32-40(2003).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, SUBUNIT, AND PATHWAY.
RC STRAIN=ATCC BAA-588 / NCTC 13252 / RB50;
RX PubMed=27218267; DOI=10.1021/acs.biochem.6b00105;
RA Hicks K.A., Yuen M.E., Zhen W.F., Gerwig T.J., Story R.W., Kopp M.C.,
RA Snider M.J.;
RT "Structural and Biochemical Characterization of 6-Hydroxynicotinic Acid 3-
RT Monooxygenase, A Novel Decarboxylative Hydroxylase Involved in Aerobic
RT Nicotinate Degradation.";
RL Biochemistry 55:3432-3446(2016).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP PATHWAY, MUTAGENESIS OF HIS-47; CYS-201; HIS-210; TYR-214; TYR-224 AND
RP HIS-301, REACTION MECHANISM, AND ACTIVE SITE.
RC STRAIN=ATCC BAA-588 / NCTC 13252 / RB50;
RX PubMed=30810301; DOI=10.1021/acs.biochem.8b00969;
RA Nakamoto K.D., Perkins S.W., Campbell R.G., Bauerle M.R., Gerwig T.J.,
RA Gerislioglu S., Wesdemiotis C., Anderson M.A., Hicks K.A., Snider M.J.;
RT "Mechanism of 6-Hydroxynicotinate 3-Monooxygenase, a Flavin-Dependent
RT Decarboxylative Hydroxylase Involved in Bacterial Nicotinic Acid
RT Degradation.";
RL Biochemistry 58:1751-1763(2019).
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 (PubMed:30810301,
CC PubMed:27218267). Is also active on the non-natural substrate 5-chloro-
CC 6-hydroxynicotinate, and is much less efficient on the substrate analog
CC 4-hydroxybenzoate (PubMed:30810301). {ECO:0000269|PubMed:27218267,
CC ECO:0000269|PubMed:30810301}.
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:30810301};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:30810301};
CC Note=Binds 1 FAD molecule per subunit. {ECO:0000250|UniProtKB:Q88FY2};
CC -!- ACTIVITY REGULATION: Competitively inhibited by 6-
CC hydroxynicotinaldehyde. {ECO:0000269|PubMed:27218267}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=85 uM for 6-hydroxynicotinate (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:27218267};
CC KM=6 uM for NADH (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:27218267};
CC KM=118 uM for 6-hydroxynicotinate (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:30810301};
CC KM=600 uM for 4-hydroxybenzoate (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:30810301};
CC KM=3.9 uM for 5-chloro-6-hydroxynicotinate (at pH 7.5 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:30810301};
CC KM=8.1 uM for NADH (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:30810301};
CC Note=kcat is 4.2 sec(-1) with 6-hydroxynicotinate as substrate (at pH
CC 7.5 and 25 degrees Celsius) (PubMed:27218267). kcat is 5.1 sec(-1)
CC with 6-hydroxynicotinate as substrate. kcat is 0.074 sec(-1) with 4-
CC hydroxybenzoate as substrate. kcat is 2.18 sec(-1) with 5-chloro-6-
CC hydroxynicotinate as substrate (at pH 7.5 and 25 degrees Celsius)
CC (PubMed:30810301). {ECO:0000269|PubMed:27218267,
CC ECO:0000269|PubMed:30810301};
CC pH dependence:
CC Optimum pH is around 7-8. {ECO:0000269|PubMed:27218267};
CC -!- PATHWAY: Cofactor degradation; nicotinate degradation.
CC {ECO:0000305|PubMed:27218267, ECO:0000305|PubMed:30810301}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:27218267}.
CC -!- SIMILARITY: Belongs to the 6-hydroxynicotinate 3-monooxygenase family.
CC {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; BX640442; CAE32275.1; -; Genomic_DNA.
DR RefSeq; WP_010926295.1; NC_002927.3.
DR AlphaFoldDB; A0A0H3LKL4; -.
DR SMR; A0A0H3LKL4; -.
DR STRING; 257310.BB1778; -.
DR EnsemblBacteria; CAE32275; CAE32275; BB1778.
DR GeneID; 56479592; -.
DR KEGG; bbr:BB1778; -.
DR eggNOG; COG0654; Bacteria.
DR HOGENOM; CLU_009665_19_5_4; -.
DR OMA; YKHMVGH; -.
DR OrthoDB; 504558at2; -.
DR UniPathway; UPA01010; -.
DR Proteomes; UP000001027; Chromosome.
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: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 Oxidoreductase; Signal.
FT SIGNAL 1..26
FT /evidence="ECO:0000255"
FT CHAIN 27..383
FT /note="6-hydroxynicotinate 3-monooxygenase"
FT /id="PRO_0000447475"
FT ACT_SITE 47
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:30810301"
FT ACT_SITE 214
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:30810301"
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 293
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT BINDING 306..307
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q88FY2"
FT MUTAGEN 47
FT /note="H->A,F: Unable to bind FAD."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 47
FT /note="H->E: Almost complete loss of 6-HNA hydroxylation
FT activity (0.02% of wild-type). Ability to oxidize NADH is
FT also greatly diminished."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 201
FT /note="C->A: No significant loss of 6-HNA hydroxylation
FT activity."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 210
FT /note="H->A: No significant loss of 6-HNA hydroxylation
FT activity."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 214
FT /note="Y->F: Great decrease in affinity for 6-
FT hydroxynicotinate. Retains ability to oxidize NADH but its
FT ability to transfer the hydroxyl group onto the organic
FT substrate is greatly diminished (0.05% of wild-type)."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 224
FT /note="Y->F: No significant loss of 6-HNA hydroxylation
FT activity."
FT /evidence="ECO:0000269|PubMed:30810301"
FT MUTAGEN 301
FT /note="H->A: 11-fold decrease in 6-HNA hydroxylation
FT activity."
FT /evidence="ECO:0000269|PubMed:30810301"
SQ SEQUENCE 383 AA; 42803 MW; 5162C880F6D65662 CRC64;
MQGKPRIAVI GAGLGGTAGA ALMARAGFNV RLYEQAPAFS RLGAGIHLGP NVMKIMRRIG
IEDELNRQGS HPDYWYSRDW QSGAELARIP LGDYAVSHYG ATYLTVHRGD FHALMTAALP
AGLLQFNKRL TRVDEDDDVV RLHFADGSVE EAEIVIGADG VNSRLREHLL GAELPKYTGY
VAHRAVFPTP LDSGSLPFDM CVKWWSDDRH MMVYFVTGKR DEIYYVTGVP EQQWDMGKSW
VPSSKAEMRA AFAGWHPTVQ ALIEATPEVS KWPLLERDPL PLWSRGRIVL LGDACHPMKP
HMAQGAAMAI EDAAMLTRIF EQTGLQDHAA AFRLYEDNRA ERASRVQRVS HDNTWLRTNE
NPDWCFGYDV YAEPLVEGRR AAA
