CLAD_PENCR
ID CLAD_PENCR Reviewed; 338 AA.
AC A0A481WNL2;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 05-JUN-2019, sequence version 1.
DT 03-AUG-2022, entry version 12.
DE RecName: Full=Clavatol oxidase claD {ECO:0000303|PubMed:30811183};
DE EC=1.14.-.- {ECO:0000269|PubMed:30811183};
DE AltName: Full=2-oxoglutarate-dependent dioxygenase claD {ECO:0000303|PubMed:30811183};
DE AltName: Full=Clavatol biosynthesis cluster protein D {ECO:0000303|PubMed:30811183};
GN Name=claD {ECO:0000303|PubMed:30811183};
OS Penicillium crustosum (Blue mold fungus).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=36656;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DISRUPTION PHENOTYPE,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RC STRAIN=PRB-2;
RX PubMed=30811183; DOI=10.1021/jacs.9b00110;
RA Fan J., Liao G., Kindinger F., Ludwig-Radtke L., Yin W.B., Li S.M.;
RT "Peniphenone and penilactone formation in Penicillium crustosum via 1,4-
RT Michael additions of ortho-quinone methide from hydroxyclavatol to gamma-
RT butyrolactones from Crustosic Acid.";
RL J. Am. Chem. Soc. 141:4225-4229(2019).
RN [2]
RP FUNCTION.
RX PubMed=31860310; DOI=10.1021/acs.joc.9b02971;
RA Liao G., Fan J., Ludwig-Radtke L., Backhaus K., Li S.M.;
RT "Increasing Structural Diversity of Natural Products by Michael Addition
RT with ortho-Quinone Methide as the Acceptor.";
RL J. Org. Chem. 85:1298-1307(2020).
CC -!- FUNCTION: 2-oxoglutarate-dependent dioxygenase; part of the cla gene
CC cluster that produces clavatol and ortho-quinone methide
CC (PubMed:30811183). The clavatol biosynthesis cluster cla and the
CC terrestric acid cluster tra are both involved in the production of
CC peniphenones and penilactones (PubMed:30811183). The non-reducing PKS
CC claF is responsible for the formation of clavatol from successive
CC condensations of 3 malonyl-CoA units, presumably with a simple acetyl-
CC CoA starter unit, and 2 methylation steps (PubMed:30811183). The
CC esterase claE probably collaborates with claF by catalyzing the
CC hydrolysis of ACP-bound acyl intermediates to free the ACP from stalled
CC intermediates (By similarity). The clavatol oxidase claD then converts
CC clavatol to hydroxyclavatol (PubMed:30811183). Spontaneous dehydration
CC of hydroxyclavatol leads to the accumulation of the highly active
CC ortho-quinone methide (PubMed:30811183, PubMed:31860310). On the other
CC hand, the PKS-NRPS hybrid traA is involved in the formation of
CC crustosic acid, with the help of traB and traD (PubMed:30811183). The
CC polyketide synthase module (PKS) of traA is responsible for the
CC synthesis of the polyketide backbone via the condensation of an acetyl-
CC CoA starter unit with 3 malonyl-CoA units (PubMed:30811183). The
CC downstream nonribosomal peptide synthetase (NRPS) module then amidates
CC the carboxyl end of the polyketide with L-malic acid (PubMed:30811183).
CC Because traA lacks a designated enoylreductase (ER) domain, the
CC required activity is provided the enoyl reductase traG (By similarity).
CC Crustosic acid undergoes decarboxylation and isomerization to the
CC terrestric acid, catalyzed by the 2-oxoglutarate-dependent dioxygenase
CC traH (PubMed:30811183). Both acids are further converted to the 2
CC gamma-butyrolactones (R)-5-methyltetronic acid and (S)-5-
CC carboxylmethyltetronic acid, with involvement of the cytochrome P450
CC monooxygenase claJ (PubMed:30811183). Spontaneous addition of the
CC methide to these gamma-butyrolactones leads to peniphenone D and
CC penilactone D, which undergo again stereospecific attacking by methide
CC to give penilactones A and B (PubMed:30811183, PubMed:31860310).
CC {ECO:0000250|UniProtKB:A0A0E0RXA7, ECO:0000250|UniProtKB:A0A161CKG1,
CC ECO:0000269|PubMed:30811183, ECO:0000269|PubMed:31860310}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + clavatol + O2 = CO2 + hydroxyclavatol +
CC succinate; Xref=Rhea:RHEA:70079, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:16810, ChEBI:CHEBI:30031,
CC ChEBI:CHEBI:188925, ChEBI:CHEBI:188926;
CC Evidence={ECO:0000269|PubMed:30811183};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70080;
CC Evidence={ECO:0000269|PubMed:30811183};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00805};
CC Note=Binds 1 Fe(2+) ion per subunit. {ECO:0000255|PROSITE-
CC ProRule:PRU00805};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.30 mM for clavatol {ECO:0000269|PubMed:30811183};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:30811183}.
CC -!- DISRUPTION PHENOTYPE: Completely abolishes the production of
CC hydroxyclavatol, as well as of the clavatol-derived compounds
CC peniphenone D, penilactone D, penilactone A, penilactone B and
CC hydroxyclavatol (PubMed:30811183). Leads to the accumulation of
CC clavatol (PubMed:30811183). {ECO:0000269|PubMed:30811183}.
CC -!- SIMILARITY: Belongs to the iron/ascorbate-dependent oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; MK360918; QBK15042.1; -; Genomic_DNA.
DR SMR; A0A481WNL2; -.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR Gene3D; 2.60.120.330; -; 1.
DR InterPro; IPR026992; DIOX_N.
DR InterPro; IPR044861; IPNS-like_FE2OG_OXY.
DR InterPro; IPR027443; IPNS-like_sf.
DR InterPro; IPR005123; Oxoglu/Fe-dep_dioxygenase.
DR Pfam; PF03171; 2OG-FeII_Oxy; 1.
DR Pfam; PF14226; DIOX_N; 1.
DR PROSITE; PS51471; FE2OG_OXY; 1.
PE 1: Evidence at protein level;
KW Dioxygenase; Iron; Metal-binding; Oxidoreductase.
FT CHAIN 1..338
FT /note="Clavatol oxidase claD"
FT /id="PRO_0000455068"
FT DOMAIN 193..299
FT /note="Fe2OG dioxygenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 222
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 224
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 280
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 290
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
SQ SEQUENCE 338 AA; 38170 MW; E338F1A04C10F179 CRC64;
MPVLSNPSFY LPLVDITPFL ENPHGAAAQD VIESVRTACK STGFFQIKGH QVPLRLQKSV
FEASARFFAL PLKNKLELDS RKTVGFRGYD VMETQSYELE FGAVQEADAL RDIKEGFFIA
TDLPPDHPHV ANGRFLQGPN VWPKPEQLAP EDFQSVLEEY YTEMQRLSHV VLSLLAATLP
YGPHVFDELE TCDPMSLLRL LHYPRGLEKQ DGKKLQLGAG EHTDFGTFTL LLQDEHPGLE
VQDSVTGEWH GVPPQEDVYI VNVADILSTM TEGDYKSSVH RVWNIKSNDR YSVVFFYDGN
LDYKVKPLRS SGQDENEEID APTIEEHVRS RLTASYAI
