HLNO_SHIS7
ID HLNO_SHIS7 Reviewed; 437 AA.
AC A0A075BSX9;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 29-OCT-2014, sequence version 1.
DT 03-AUG-2022, entry version 45.
DE RecName: Full=(S)-6-hydroxynicotine oxidase {ECO:0000303|PubMed:25002425};
DE Short=(S)-6HN oxidase {ECO:0000303|PubMed:25002425};
DE EC=1.5.3.5 {ECO:0000269|PubMed:25002425};
DE AltName: Full=6-hydroxy-L-nicotine oxidase {ECO:0000305};
DE Short=6-HLNO {ECO:0000305};
DE AltName: Full=L-hydroxynicotine oxidase {ECO:0000305};
DE Short=LHNO {ECO:0000305};
GN Name=nctB {ECO:0000303|PubMed:25002425};
GN ORFNames=shn_30305 {ECO:0000312|EMBL:ANH08442.1};
OS Shinella sp. (strain HZN7).
OG Plasmid pShin-05.
OC Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC Rhizobiaceae; Shinella; unclassified Shinella.
OX NCBI_TaxID=879274;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND DISRUPTION
RP PHENOTYPE.
RC STRAIN=HZN7;
RX PubMed=25002425; DOI=10.1128/aem.01312-14;
RA Qiu J., Wei Y., Ma Y., Wen R., Wen Y., Liu W.;
RT "A novel (S)-6-hydroxynicotine oxidase gene from Shinella sp. strain
RT HZN7.";
RL Appl. Environ. Microbiol. 80:5552-5560(2014).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=HZN7; PLASMID=pShin-05;
RX PubMed=27625640; DOI=10.3389/fmicb.2016.01348;
RA Qiu J., Yang Y., Zhang J., Wang H., Ma Y., He J., Lu Z.;
RT "The complete genome sequence of the nicotine-degrading bacterium Shinella
RT sp. HZN7.";
RL Front. Microbiol. 7:1348-1348(2016).
RN [3]
RP FUNCTION IN NORNICOTINE DEGRADATION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND DISRUPTION PHENOTYPE.
RX PubMed=27568381; DOI=10.1007/s00253-016-7805-0;
RA Qiu J., Li N., Lu Z., Yang Y., Ma Y., Niu L., He J., Liu W.;
RT "Conversion of nornicotine to 6-hydroxy-nornicotine and 6-hydroxy-myosmine
RT by Shinella sp. strain HZN7.";
RL Appl. Microbiol. Biotechnol. 100:10019-10029(2016).
RN [4] {ECO:0007744|PDB:6CR0}
RP X-RAY CRYSTALLOGRAPHY (1.55 ANGSTROMS) IN COMPLEX WITH FAD.
RA Richter M.;
RT "1.55 A resolution structure of (S)-6-hydroxynicotine oxidase from Shinella
RT HZN7.";
RL Submitted (MAR-2018) to the PDB data bank.
CC -!- FUNCTION: Involved in the degradation of L-nicotine (PubMed:25002425).
CC Catalyzes the oxidation of (S)-6-hydroxynicotine (6-hydroxy-L-nicotine)
CC to 6-hydroxypseudooxynicotine (PubMed:25002425). Oxidation of the
CC pyrrolidine ring of (S)-6-hydroxynicotine leads to the formation of the
CC optically inactive 6-hydroxy-N-methylmyosmine, which hydrolyzes
CC spontaneously to 6-hydroxypseudooxynicotine (PubMed:25002425). Acts
CC with absolute stereospecificity on the L-form of 6-hydroxynicotine
CC (PubMed:25002425). Also involved in the degradation of nornicotine, and
CC catalyzes the oxidation of 6-hydroxynornicotine to 6-hydroxymyosmine,
CC which hydrolyzes to 6-hydroxypseudooxynornicotine (PubMed:27568381). In
CC vitro, converts (S)-nicotine into N-methylmyosmine, which spontaneously
CC hydrolyzes spontaneously into pseudooxynicotine, but catalytic
CC efficiency is about 1900-fold higher with (S)-6-hydroxynicotine
CC (PubMed:25002425). {ECO:0000269|PubMed:25002425,
CC ECO:0000269|PubMed:27568381}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-6-hydroxynicotine + H2O + O2 = 6-hydroxypseudooxynicotine
CC + H2O2; Xref=Rhea:RHEA:11880, ChEBI:CHEBI:15377, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16240, ChEBI:CHEBI:58182, ChEBI:CHEBI:58682; EC=1.5.3.5;
CC Evidence={ECO:0000269|PubMed:25002425};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:11881;
CC Evidence={ECO:0000269|PubMed:25002425};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-6-hydroxynicotine + O2 = 6-hydroxy-N-methylmyosmine +
CC H2O2; Xref=Rhea:RHEA:46976, ChEBI:CHEBI:15379, ChEBI:CHEBI:16240,
CC ChEBI:CHEBI:58182, ChEBI:CHEBI:87164;
CC Evidence={ECO:0000269|PubMed:25002425};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:46977;
CC Evidence={ECO:0000269|PubMed:25002425};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:25002425};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:25002425};
CC -!- ACTIVITY REGULATION: Partially inhibited by Co(2+) or Zn(2+) and
CC significantly inhibited by Ag(+), Cu(2+) and Hg(2+).
CC {ECO:0000269|PubMed:25002425}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.019 mM for (S)-6-hydroxynicotine {ECO:0000269|PubMed:25002425};
CC KM=0.033 mM for 6-hydroxynornicotine {ECO:0000269|PubMed:27568381};
CC KM=2.03 mM for nicotine {ECO:0000269|PubMed:25002425};
CC Note=kcat is 7.3 sec(-1) with (S)-6-hydroxynicotine as substrate.
CC kcat is 0.396 sec(-1) with nicotine as substrate (PubMed:25002425).
CC kcat is 5.9 sec(-1) with 6-hydroxynornicotine as substrate
CC (PubMed:27568381). {ECO:0000269|PubMed:25002425,
CC ECO:0000269|PubMed:27568381};
CC pH dependence:
CC Optimum pH is 7.0. Stable between pH 6.0 and 9.8.
CC {ECO:0000269|PubMed:25002425};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius. Stable at temperatures
CC lower than 50 degrees Celsius. {ECO:0000269|PubMed:25002425};
CC -!- PATHWAY: Alkaloid degradation; nicotine degradation; 6-
CC hydroxypseudooxynicotine from nicotine (S-isomer route): step 2/2.
CC {ECO:0000305}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:25002425}.
CC -!- DISRUPTION PHENOTYPE: Disruption of the gene abolishes the ability of
CC the mutant to degrade (S)-6-hydroxynicotine.
CC {ECO:0000269|PubMed:25002425}.
CC -!- SIMILARITY: Belongs to the flavin monoamine oxidase family.
CC {ECO:0000305}.
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DR EMBL; KF306095; AGS16700.1; -; Genomic_DNA.
DR EMBL; CP015741; ANH08442.1; -; Genomic_DNA.
DR RefSeq; WP_064334256.1; NZ_CP015741.1.
DR PDB; 6CR0; X-ray; 1.55 A; A=1-437.
DR PDBsum; 6CR0; -.
DR SMR; A0A075BSX9; -.
DR EnsemblBacteria; ANH08442; ANH08442; shn_30305.
DR KEGG; shz:shn_30305; -.
DR OrthoDB; 1442136at2; -.
DR BioCyc; MetaCyc:MON-17710; -.
DR BRENDA; 1.5.3.5; 15268.
DR UniPathway; UPA00106; UER00919.
DR Proteomes; UP000077495; Plasmid pShin-05.
DR GO; GO:0018531; F:(S)-6-hydroxynicotine oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0009820; P:alkaloid metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0019608; P:nicotine catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.50.50.60; -; 3.
DR InterPro; IPR002937; Amino_oxidase.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR001613; Flavin_amine_oxidase.
DR Pfam; PF01593; Amino_oxidase; 1.
DR PRINTS; PR00757; AMINEOXDASEF.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; FAD; Flavoprotein; Nucleotide-binding;
KW Oxidoreductase; Plasmid.
FT CHAIN 1..437
FT /note="(S)-6-hydroxynicotine oxidase"
FT /id="PRO_0000455451"
FT BINDING 16
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 35..37
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 43
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 57..60
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 231
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 405
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
FT BINDING 413..415
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|Ref.4, ECO:0007744|PDB:6CR0"
SQ SEQUENCE 437 AA; 48736 MW; 99FC8795861E2EB1 CRC64;
MTEKIYDAIV VGAGFSGLVA ARELSAQGRS VLIIEARHRL GGRTHVVNFL GRPVEIGGAG
VHWCQPHVFA EMQRYGFGFK EAPLADLDKA YMVFADGQKI DVPPATFDEE YTTAFEKFCS
RSRELFPRPY SPLDNHEVSN LDGVSARDHL ESLGLNELQL ASMNAELTLY GGAPTTELSY
PSFVKFHALA SWDTITFTDS EKRYHVQGGT NALCQAIFDD CRADSEFGVP VEAVAQTDNG
VTVTLADKRV FRALTCVLTL PTKVYADVRF EPPLPPEKRA FIEHAEMADG AELYVHVRQN
LGNTFTFCDD PNPFNAVQTY AYDDELGTIL KITIGRQSLI NLENFDAIAA EIRKIHGDVE
VLEALPYNWA MDEYARTSYP AMRKGWFSRY KDMAKPENRL FFAGSATADG WHEYIDGAIE
SGIRVGREIR HFMKATA
