DHPON_PAENI
ID DHPON_PAENI Reviewed; 367 AA.
AC Q93NG6;
DT 03-APR-2007, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 25-MAY-2022, entry version 81.
DE RecName: Full=2,6-dihydropseudooxynicotine hydrolase;
DE EC=3.7.1.19;
OS Paenarthrobacter nicotinovorans (Arthrobacter nicotinovorans).
OG Plasmid pAO1 {ECO:0000312|EMBL:CAD47941.1}.
OC Bacteria; Actinobacteria; Micrococcales; Micrococcaceae; Paenarthrobacter.
OX NCBI_TaxID=29320;
RN [1] {ECO:0000305, ECO:0000312|EMBL:AAK64252.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=11514508; DOI=10.1128/jb.183.18.5262-5267.2001;
RA Baitsch D., Sandu C., Brandsch R., Igloi G.L.;
RT "Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in
RT degradation of the plant alkaloid nicotine: cloning, purification, and
RT characterization of 2,6-dihydroxypyridine 3-hydroxylase.";
RL J. Bacteriol. 183:5262-5267(2001).
RN [2] {ECO:0000312|EMBL:CAD47941.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=12618462; DOI=10.1128/jb.185.6.1976-1986.2003;
RA Igloi G.L., Brandsch R.;
RT "Sequence of the 165-kilobase catabolic plasmid pAO1 from Arthrobacter
RT nicotinovorans and identification of a pAO1-dependent nicotine uptake
RT system.";
RL J. Bacteriol. 185:1976-1986(2003).
RN [3] {ECO:0000305}
RP PATHWAY.
RX PubMed=5835946; DOI=10.1016/s0021-9258(18)97197-8;
RA Gherna R.L., Richardson S.H., Rittenberg S.C.;
RT "The bacterial oxidation of nicotine. VI. The metabolism of 2,6-
RT dihydroxypseudooxynicotine.";
RL J. Biol. Chem. 240:3669-3674(1965).
RN [4] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=16321959; DOI=10.1128/jb.187.24.8516-8519.2005;
RA Sachelaru P., Schiltz E., Igloi G.L., Brandsch R.;
RT "An alpha/beta-fold C--C bond hydrolase is involved in a central step of
RT nicotine catabolism by Arthrobacter nicotinovorans.";
RL J. Bacteriol. 187:8516-8519(2005).
RN [5] {ECO:0000305}
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP PATHWAY, SUBUNIT, ACTIVE SITE, AND MUTAGENESIS OF ARG-18; ASP-22; GLU-148;
RP SER-149; GLU-153; ARG-216; SER-217; ASP-300; HIS-329; CYS-330 AND HIS-332.
RX PubMed=17275835; DOI=10.1016/j.jmb.2006.12.068;
RA Schleberger C., Sachelaru P., Brandsch R., Schulz G.E.;
RT "Structure and action of a C-C bond cleaving alpha/beta-hydrolase involved
RT in nicotine degradation.";
RL J. Mol. Biol. 367:409-418(2007).
CC -!- FUNCTION: L-nicotine is used as a growth substrate. Plays a role in
CC nicotine catabolism by cleaving a C-C bond in 2,6-
CC dihydroxypseudooxynicotine. {ECO:0000269|PubMed:16321959,
CC ECO:0000269|PubMed:17275835}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2,6-dihydroxypseudooxynicotine + H2O = 2,6-dihydroxypyridine +
CC 4-(methylamino)butanoate + H(+); Xref=Rhea:RHEA:34167,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:17681,
CC ChEBI:CHEBI:66882, ChEBI:CHEBI:66944; EC=3.7.1.19;
CC Evidence={ECO:0000269|PubMed:16321959, ECO:0000269|PubMed:17275835};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=6 uM for 2,6-dihydropseudooxynicotine
CC {ECO:0000269|PubMed:16321959};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:16321959};
CC -!- PATHWAY: Alkaloid degradation; nicotine degradation; 2,6-
CC dihydroxypyridine and 4-(methylamino)butanoate from 6-
CC hydroxypseudooxynicotine: step 2/2. {ECO:0000269|PubMed:16321959,
CC ECO:0000269|PubMed:17275835, ECO:0000269|PubMed:5835946}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:17275835}.
CC -!- SIMILARITY: Belongs to the AB hydrolase superfamily. {ECO:0000305}.
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DR EMBL; AF373840; AAK64252.1; -; Genomic_DNA.
DR EMBL; AJ507836; CAD47941.1; -; Genomic_DNA.
DR RefSeq; WP_016359452.1; NC_021229.1.
DR RefSeq; YP_007988767.1; NC_021229.1.
DR PDB; 2JBW; X-ray; 2.10 A; A/B/C/D=1-365.
DR PDBsum; 2JBW; -.
DR AlphaFoldDB; Q93NG6; -.
DR SMR; Q93NG6; -.
DR ESTHER; artni-Q93NG6; Duf_1100-S.
DR MEROPS; S09.A77; -.
DR KEGG; ag:CAD47941; -.
DR BRENDA; 3.7.1.19; 449.
DR SABIO-RK; Q93NG6; -.
DR UniPathway; UPA00106; UER00490.
DR EvolutionaryTrace; Q93NG6; -.
DR GO; GO:0016787; F:hydrolase activity; 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; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR010520; FrsA-like.
DR Pfam; PF06500; FrsA-like; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alkaloid metabolism; Hydrolase; Plasmid.
FT CHAIN 1..367
FT /note="2,6-dihydropseudooxynicotine hydrolase"
FT /id="PRO_0000283735"
FT ACT_SITE 148
FT /evidence="ECO:0000255, ECO:0000303|PubMed:17275835"
FT ACT_SITE 217
FT /evidence="ECO:0000269|PubMed:17275835"
FT ACT_SITE 300
FT /evidence="ECO:0000269|PubMed:17275835"
FT ACT_SITE 329
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 18
FT /note="R->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 22
FT /note="D->A: 91% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 148
FT /note="E->A: 98% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 149
FT /note="S->A: 84% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 153
FT /note="E->A: 32% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 216
FT /note="R->A: 92% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 217
FT /note="S->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 300
FT /note="D->A: 94% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 329
FT /note="H->A: 92% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 330
FT /note="C->A: 5% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT MUTAGEN 332
FT /note="H->A: 58% loss of activity."
FT /evidence="ECO:0000269|PubMed:17275835"
FT HELIX 9..13
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 16..22
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 26..33
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 41..61
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 65..83
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 90..105
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 106..108
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 109..111
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 113..120
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 123..130
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 133..135
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 138..144
FT /evidence="ECO:0007829|PDB:2JBW"
FT TURN 151..153
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 155..163
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 167..171
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 177..179
FT /evidence="ECO:0007829|PDB:2JBW"
FT TURN 180..182
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 189..202
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 206..216
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 218..229
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 235..240
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 248..250
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 253..262
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 268..278
FT /evidence="ECO:0007829|PDB:2JBW"
FT TURN 282..284
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 285..287
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 292..297
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 300..302
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 305..313
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 316..318
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 319..324
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 329..334
FT /evidence="ECO:0007829|PDB:2JBW"
FT HELIX 337..350
FT /evidence="ECO:0007829|PDB:2JBW"
FT STRAND 360..362
FT /evidence="ECO:0007829|PDB:2JBW"
SQ SEQUENCE 367 AA; 40995 MW; 1A5D31A7DFC4C8E3 CRC64;
MTVTSQVKPE DEMLNWGRLI LDGVSYSDMV GARDRPKEIT WFDYWMSLAN EYEQEAERKV
ALGHDLSAGE LLMSAALCAQ YAQFLWFDER RQKGQARKVE LYQKAAPLLS PPAERHELVV
DGIPMPVYVR IPEGPGPHPA VIMLGGLEST KEESFQMENL VLDRGMATAT FDGPGQGEMF
EYKRIAGDYE KYTSAVVDLL TKLEAIRNDA IGVLGRSLGG NYALKSAACE PRLAACISWG
GFSDLDYWDL ETPLTKESWK YVSKVDTLEE ARLHVHAALE TRDVLSQIAC PTYILHGVHD
EVPLSFVDTV LELVPAEHLN LVVEKDGDHC CHNLGIRPRL EMADWLYDVL VAGKKVAPTM
KGWPLNG
