OTEMO_PSEPU
ID OTEMO_PSEPU Reviewed; 545 AA.
AC H3JQW0; M5AXJ6;
DT 01-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 18-APR-2012, sequence version 1.
DT 03-AUG-2022, entry version 32.
DE RecName: Full=2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetyl-CoA monooxygenase;
DE Short=OTEMO;
DE EC=1.14.13.160;
DE AltName: Full=(2,2,3-trimethyl-5-oxocyclopent-3-enyl)acetyl-CoA 1,5-monooxygenase;
DE AltName: Full=MO2;
GN Name=otemo; Synonyms=camG {ECO:0000303|PubMed:23524667};
OS Pseudomonas putida (Arthrobacter siderocapsulatus).
OG Plasmid CAM.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=303;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE SPECIFICITY.
RC STRAIN=ATCC 17453 / DSM 50198 / JCM 6157 / NCIMB 10007 / NRRL B-4067 /
RC Stanier 77 / Biotype A;
RX PubMed=22286514; DOI=10.1007/s00253-011-3859-1;
RA Kadow M., Loschinski K., Sass S., Schmidt M., Bornscheuer U.T.;
RT "Completing the series of BVMOs involved in camphor metabolism of
RT Pseudomonas putida NCIMB 10007 by identification of the two missing genes,
RT their functional expression in E. coli, and biochemical characterization.";
RL Appl. Microbiol. Biotechnol. 96:419-429(2012).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 17453 / DSM 50198 / JCM 6157 / NCIMB 10007 / NRRL B-4067 /
RC Stanier 77 / Biotype A;
RX PubMed=23524667; DOI=10.1128/aem.03958-12;
RA Iwaki H., Grosse S., Bergeron H., Leisch H., Morley K., Hasegawa Y.,
RA Lau P.C.K.;
RT "Camphor pathway redux: functional recombinant expression of 2,5- and 3,6-
RT diketocamphane monooxygenases of Pseudomonas putida ATCC 17453 with their
RT cognate flavin reductase catalyzing Baeyer-Villiger reactions.";
RL Appl. Environ. Microbiol. 79:3282-3293(2013).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, INDUCTION, BIOPHYSICOCHEMICAL PROPERTIES,
RP COFACTOR, AND SUBUNIT.
RX PubMed=6848481; DOI=10.1128/jb.153.1.140-152.1983;
RA Ougham H.J., Taylor D.G., Trudgill P.W.;
RT "Camphor revisited: involvement of a unique monooxygenase in metabolism of
RT 2-oxo-delta 3-4,5,5-trimethylcyclopentenylacetic acid by Pseudomonas
RT putida.";
RL J. Bacteriol. 153:140-152(1983).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.04 ANGSTROMS) IN COMPLEX WITH FAD AND NADP,
RP FUNCTION, MUTAGENESIS OF TYR-53; ASP-59 AND ARG-337, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, COFACTOR, AND SUBUNIT.
RC STRAIN=ATCC 17453 / DSM 50198 / JCM 6157 / NCIMB 10007 / NRRL B-4067 /
RC Stanier 77 / Biotype A;
RX PubMed=22267661; DOI=10.1128/aem.07694-11;
RA Leisch H., Shi R., Grosse S., Morley K., Bergeron H., Cygler M., Iwaki H.,
RA Hasegawa Y., Lau P.C.K.;
RT "Cloning, Baeyer-Villiger biooxidations, and structures of the camphor
RT pathway 2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetyl-coenzyme A
RT monooxygenase of Pseudomonas putida ATCC 17453.";
RL Appl. Environ. Microbiol. 78:2200-2212(2012).
CC -!- FUNCTION: Involved in the degradation of (+)-camphor. Catalyzes the
CC lactonization of 2-oxo-delta(3)-4,5, 5-trimethylcyclopentenylacetyl-CoA
CC (OT-CoA), a key intermediate in the metabolism of camphor. 2-
CC Oxocyclopentyl ethyl acetate is also a good substrate, as is 2-
CC oxocyclohexyl ethyl acetate and methyl-substituted cyclohexanones, but
CC free acid is a poor substrate. {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:22286514, ECO:0000269|PubMed:6848481}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[(1R)-2,2,3-trimethyl-5-oxocyclopent-3-enyl]acetyl-CoA + H(+)
CC + NADPH + O2 = [(2R)-3,3,4-trimethyl-6-oxo-3,6-dihydro-1H-pyran-2-
CC yl]acetyl-CoA + H2O + NADP(+); Xref=Rhea:RHEA:33015,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:64784,
CC ChEBI:CHEBI:64785; EC=1.14.13.160;
CC Evidence={ECO:0000269|PubMed:22286514, ECO:0000269|PubMed:6848481};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:6848481};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:6848481};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.018 mM for OT-CoA (at pH 9 and at 25 degrees Celsius)
CC {ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:22286514,
CC ECO:0000269|PubMed:6848481};
CC KM=0.032 mM for 2-n-hexyl cyclopentanone (at pH 9 and at 25 degrees
CC Celsius) {ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:22286514,
CC ECO:0000269|PubMed:6848481};
CC KM=0.16 mM for 2-oxocyclohexyl ethyl acetate (at pH 9 and at 25
CC degrees Celsius) {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:22286514, ECO:0000269|PubMed:6848481};
CC KM=0.31 mM for 2-oxocyclopentyl ethyl acetate (at pH 9 and at 25
CC degrees Celsius) {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:22286514, ECO:0000269|PubMed:6848481};
CC KM=0.6 mM for 4-methyl cyclohexanone (at pH 9 and at 25 degrees
CC Celsius) {ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:22286514,
CC ECO:0000269|PubMed:6848481};
CC KM=1.4 mM for 2-methyl cyclohexanone (at pH 9 and at 25 degrees
CC Celsius) {ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:22286514,
CC ECO:0000269|PubMed:6848481};
CC pH dependence:
CC Optimum pH is 9. {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:22286514, ECO:0000269|PubMed:6848481};
CC Temperature dependence:
CC Optimum temperature is 20 degrees Celsius.
CC {ECO:0000269|PubMed:22267661, ECO:0000269|PubMed:22286514,
CC ECO:0000269|PubMed:6848481};
CC -!- PATHWAY: Terpene metabolism; (R)-camphor degradation.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:22267661,
CC ECO:0000269|PubMed:6848481}.
CC -!- INDUCTION: Induced by (+)-camphor and 2-oxo-delta(3)-4,5,5-
CC trimethylcyclopentenylacetic acid. {ECO:0000269|PubMed:6848481}.
CC -!- SIMILARITY: Belongs to the FAD-binding monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; JQ034405; AEZ35248.1; -; Genomic_DNA.
DR EMBL; AB771747; BAN13280.1; -; Genomic_DNA.
DR PDB; 3UOV; X-ray; 2.04 A; A/B=1-545.
DR PDB; 3UOX; X-ray; 1.96 A; A/B=1-545.
DR PDB; 3UOY; X-ray; 2.00 A; A/B=1-545.
DR PDB; 3UOZ; X-ray; 2.41 A; A/B=1-545.
DR PDB; 3UP4; X-ray; 2.80 A; A/B=1-545.
DR PDB; 3UP5; X-ray; 2.45 A; A/B=1-545.
DR PDBsum; 3UOV; -.
DR PDBsum; 3UOX; -.
DR PDBsum; 3UOY; -.
DR PDBsum; 3UOZ; -.
DR PDBsum; 3UP4; -.
DR PDBsum; 3UP5; -.
DR AlphaFoldDB; H3JQW0; -.
DR SMR; H3JQW0; -.
DR KEGG; ag:AEZ35248; -.
DR KEGG; ag:BAN13280; -.
DR BRENDA; 1.14.13.160; 5092.
DR UniPathway; UPA00719; -.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0019383; P:(+)-camphor catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR023753; FAD/NAD-binding_dom.
DR Pfam; PF07992; Pyr_redox_2; 1.
DR SUPFAM; SSF51905; SSF51905; 2.
PE 1: Evidence at protein level;
KW 3D-structure; FAD; Flavoprotein; Monooxygenase; NADP; Oxidoreductase;
KW Plasmid.
FT CHAIN 1..545
FT /note="2-oxo-Delta(3)-4,5,5-trimethylcyclopentenylacetyl-
FT CoA monooxygenase"
FT /id="PRO_0000422223"
FT BINDING 20
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 39
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 47..50
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 57..59
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 59..60
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 65
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 112
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 193..199
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 216..217
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 446
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:22267661"
FT BINDING 501
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:22267661"
FT SITE 337
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 53
FT /note="Y->A: Loss of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 53
FT /note="Y->F: Retains 32% of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 59
FT /note="D->A: Loss of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 59
FT /note="D->N: Loss of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 337
FT /note="R->A: Loss of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT MUTAGEN 337
FT /note="R->K: Loss of monooxygenase activity."
FT /evidence="ECO:0000269|PubMed:22267661"
FT STRAND 9..15
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 19..30
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 35..38
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 40..44
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 47..50
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 62..65
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 67..70
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 80..83
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 86..100
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 103..105
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 106..109
FT /evidence="ECO:0007829|PDB:3UOZ"
FT STRAND 112..118
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 119..121
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 123..128
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 129..131
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 132..141
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 157..159
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 162..166
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 167..169
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 174..176
FT /evidence="ECO:0007829|PDB:3UOV"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:3UOZ"
FT STRAND 187..191
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 195..204
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 205..207
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 208..217
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 231..239
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 241..248
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 251..256
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 260..262
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 264..266
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 269..281
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 282..284
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 286..289
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 290..292
FT /evidence="ECO:0007829|PDB:3UOZ"
FT TURN 293..297
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 299..316
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 320..325
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 329..331
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 340..343
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 344..347
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 353..357
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 358..360
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 363..367
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 370..375
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 377..379
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 381..385
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 390..393
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 395..398
FT /evidence="ECO:0007829|PDB:3UOY"
FT STRAND 399..402
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 404..406
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 409..412
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 413..415
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 421..423
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 431..433
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 437..439
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 441..443
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 446..467
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 472..474
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 476..491
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 495..498
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 500..502
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 503..507
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 509..511
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 513..519
FT /evidence="ECO:0007829|PDB:3UOX"
FT HELIX 524..536
FT /evidence="ECO:0007829|PDB:3UOX"
FT TURN 537..541
FT /evidence="ECO:0007829|PDB:3UOX"
FT STRAND 542..544
FT /evidence="ECO:0007829|PDB:3UOX"
SQ SEQUENCE 545 AA; 61374 MW; 4ABB95E87E817D8D CRC64;
MSNRAKSPAL DAVVIGAGVT GIYQAFLINQ AGMKVLGIEA GEDVGGTWYW NRYPGCRLDT
ESYAYGYFAL KGIIPEWEWS ENFASQPEML RYVNRAADAM DVRKHYRFNT RVTAARYVEN
DRLWEVTLDN EEVVTCRFLI SATGPLSASR MPDIKGIDSF KGESFHSSRW PTDAEGAPKG
VDFTGKRVGV IGTGATGVQI IPIAAETAKE LYVFQRTPNW CTPLGNSPMS KEKMDSLRNR
YPTILEYVKS TDTAFPYHRD PRKGTDVSES ERDAFFEELY RQPGYGIWLS GFRDLLLNKE
SNKFLADFVA KKIRQRVKDP VVAEKLIPKD HPFGAKRVPM ETNYYETYNR DNVHLVDIRE
APIQEVTPEG IKTADAAYDL DVIIYATGFD AVTGSLDRID IRGKDNVRLI DAWAEGPSTY
LGLQARGFPN FFTLVGPHNG STFCNVGVCG GLQAEWVLRM ISYMKDNGFT YSEPTQAAEN
RWTEEVYADF SRTLLAEANA WWVKTTTKPD GSVVRRTLVH VSGGPEYRKR CEQVAYNNYN
GFELA
