IPNA_PENCH
ID IPNA_PENCH Reviewed; 331 AA.
AC P08703;
DT 01-JAN-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1988, sequence version 1.
DT 03-AUG-2022, entry version 109.
DE RecName: Full=Isopenicillin N synthase {ECO:0000303|PubMed:3104145};
DE Short=IPNS {ECO:0000303|PubMed:3104145};
DE EC=1.21.3.1 {ECO:0000269|PubMed:1368505, ECO:0000269|PubMed:1369045, ECO:0000269|PubMed:1588566};
DE AltName: Full=Isopenicillin cyclase {ECO:0000305};
DE AltName: Full=Penicillin biosynthetis cluster protein ipnA {ECO:0000303|PubMed:1368505};
GN Name=ipnA {ECO:0000303|PubMed:3104145}; Synonyms=pcbC;
OS Penicillium chrysogenum (Penicillium notatum).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium;
OC Penicillium chrysogenum species complex.
OX NCBI_TaxID=5076;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=23X-80-269-37-2;
RX PubMed=3104145; DOI=10.1016/0378-1119(86)90084-3;
RA Carr L.G., Skatrud P.L., Scheetz M.E. II, Queener S.W., Ingolia T.D.;
RT "Cloning and expression of the isopenicillin N synthetase gene from
RT Penicillium chrysogenum.";
RL Gene 48:257-266(1986).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=AS-P-78;
RX PubMed=2499766; DOI=10.1007/bf00332235;
RA Barrredo J.L., Cantoral J.M., Alvarez E., Diez B., Martin J.F.;
RT "Cloning, sequence analysis and transcriptional study of the isopenicillin
RT N synthase of Penicillium chrysogenum AS-P-78.";
RL Mol. Gen. Genet. 216:91-98(1989).
RN [3]
RP INDUCTION.
RX PubMed=3096965; DOI=10.1128/jb.168.2.947-952.1986;
RA Revilla G., Ramos F.R., Lopez-Nieto M.J., Alvarez E., Martin J.F.;
RT "Glucose represses formation of delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-
RT valine and isopenicillin N synthase but not penicillin acyltransferase in
RT Penicillium chrysogenum.";
RL J. Bacteriol. 168:947-952(1986).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=1368505; DOI=10.1038/nbt0190-39;
RA Smith D.J., Burnham M.K., Edwards J., Earl A.J., Turner G.;
RT "Cloning and heterologous expression of the penicillin biosynthetic gene
RT cluster from penicillum chrysogenum.";
RL Biotechnology (N.Y.) 8:39-41(1990).
RN [5]
RP SUBCELLULAR LOCATION.
RX PubMed=1899377; DOI=10.1002/j.1460-2075.1991.tb07971.x;
RA Mueller W.H., van der Krift T.P., Krouwer A.J., Woesten H.A.,
RA van der Voort L.H., Smaal E.B., Verkleij A.J.;
RT "Localization of the pathway of the penicillin biosynthesis in Penicillium
RT chrysogenum.";
RL EMBO J. 10:489-495(1991).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=1369045; DOI=10.1002/jctb.280550306;
RA Bainbridge Z.A., Scott R.I., Perry D.;
RT "Oxygen utilisation by isopenicillin N synthase from Penicillium
RT chrysogenum.";
RL J. Chem. Technol. Biotechnol. 55:233-238(1992).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND PATHWAY.
RX PubMed=1588566; DOI=10.1021/jm00088a028;
RA Huffman G.W., Gesellchen P.D., Turner J.R., Rothenberger R.B.,
RA Osborne H.E., Miller F.D., Chapman J.L., Queener S.W.;
RT "Substrate specificity of isopenicillin N synthase.";
RL J. Med. Chem. 35:1897-1914(1992).
RN [8]
RP INDUCTION.
RX PubMed=8736532; DOI=10.1046/j.1365-2958.1996.5421065.x;
RA Suarez T., Penalva M.A.;
RT "Characterization of a Penicillium chrysogenum gene encoding a PacC
RT transcription factor and its binding sites in the divergent pcbAB-pcbC
RT promoter of the penicillin biosynthetic cluster.";
RL Mol. Microbiol. 20:529-540(1996).
RN [9]
RP SUBCELLULAR LOCATION.
RX PubMed=12223189; DOI=10.1016/s1087-1845(02)00036-1;
RA van der Lende T.R., van de Kamp M., Berg M., Sjollema K., Bovenberg R.A.,
RA Veenhuis M., Konings W.N., Driessen A.J.;
RT "delta-(L-alpha-Aminoadipyl)-L-cysteinyl-D-valine synthetase, that mediates
RT the first committed step in penicillin biosynthesis, is a cytosolic
RT enzyme.";
RL Fungal Genet. Biol. 37:49-55(2002).
RN [10]
RP INDUCTION.
RX PubMed=22960281; DOI=10.1016/j.fgb.2012.08.002;
RA Dominguez-Santos R., Martin J.F., Kosalkova K., Prieto C., Ullan R.V.,
RA Garcia-Estrada C.;
RT "The regulatory factor PcRFX1 controls the expression of the three genes of
RT beta-lactam biosynthesis in Penicillium chrysogenum.";
RL Fungal Genet. Biol. 49:866-881(2012).
CC -!- FUNCTION: Isopenicillin N synthase; part of the gene cluster that
CC mediates the biosynthesis of penicillin, the world's most important
CC antibiotic (PubMed:1368505, PubMed:1369045, PubMed:1588566). IpnA
CC catalyzes the cyclization of the tripeptide N-[(5S)-5-amino-5-
CC carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) to form
CC isopenicillin N (IPN) that contains the beta-lactam nucleus
CC (PubMed:1368505, PubMed:1369045, PubMed:1588566). The penicillin
CC biosynthesis occurs via 3 enzymatic steps, the first corresponding to
CC the production of the tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-
CC cysteinyl-D-valine (LLD-ACV or ACV) by the NRPS acvA. The tripeptide
CC ACV is then cyclized to isopenicillin N (IPN) by the isopenicillin N
CC synthase ipnA that forms the beta-lactam nucleus. Finally, the alpha-
CC aminoadipyl side chain is exchanged for phenylacetic acid by the
CC isopenicillin N acyltransferase aatA to yield penicillin in the
CC peroxisomal matrix (PubMed:1368505) (Probable).
CC {ECO:0000269|PubMed:1368505, ECO:0000269|PubMed:1369045,
CC ECO:0000269|PubMed:1588566, ECO:0000305|PubMed:1368505}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
CC = 2 H2O + isopenicillin N; Xref=Rhea:RHEA:22428, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:58399, ChEBI:CHEBI:58572; EC=1.21.3.1;
CC Evidence={ECO:0000269|PubMed:1368505, ECO:0000269|PubMed:1369045,
CC ECO:0000269|PubMed:1588566};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:22429;
CC Evidence={ECO:0000269|PubMed:1368505, ECO:0000269|PubMed:1369045,
CC ECO:0000269|PubMed:1588566};
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 -!- PATHWAY: Antibiotic biosynthesis; penicillin G biosynthesis; penicillin
CC G from L-alpha-aminoadipate and L-cysteine and L-valine: step 2/3.
CC {ECO:0000269|PubMed:1368505, ECO:0000269|PubMed:1369045,
CC ECO:0000269|PubMed:1588566}.
CC -!- SUBUNIT: Monomer. {ECO:0000250|UniProtKB:P05189}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol {ECO:0000269|PubMed:12223189,
CC ECO:0000269|PubMed:1899377}.
CC -!- INDUCTION: Expression is repressed by glucose (PubMed:3096965). The
CC transcription factor rfx1 controls penicillin biosynthesis through the
CC regulation of the acvA, ipnA and aatA transcription (PubMed:22960281).
CC Expression is also controlled by the transcription factor pacC that
CC specifically recognizes the 5'-GCCARG-3' sequence in the promoter
CC (PubMed:8736532). {ECO:0000269|PubMed:22960281,
CC ECO:0000269|PubMed:3096965, ECO:0000269|PubMed:8736532}.
CC -!- SIMILARITY: Belongs to the iron/ascorbate-dependent oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; M15083; AAA33696.1; -; Genomic_DNA.
DR EMBL; X17436; CAA35480.1; -; Genomic_DNA.
DR PIR; A26467; A26467.
DR PIR; S04441; S04441.
DR AlphaFoldDB; P08703; -.
DR SMR; P08703; -.
DR ChEMBL; CHEMBL3158; -.
DR BRENDA; 1.21.3.1; 4606.
DR UniPathway; UPA00149; UER00240.
DR GO; GO:0005829; C:cytosol; IDA:GO_Central.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0016216; F:isopenicillin-N synthase activity; IDA:GO_Central.
DR GO; GO:0042318; P:penicillin biosynthetic process; IDA:GO_Central.
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; IPR002057; Isopenicillin-N_synth_CS.
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.
DR PROSITE; PS00185; IPNS_1; 1.
DR PROSITE; PS00186; IPNS_2; 1.
PE 1: Evidence at protein level;
KW Antibiotic biosynthesis; Cytoplasm; Dioxygenase; Iron; Metal-binding;
KW Oxidoreductase.
FT CHAIN 1..331
FT /note="Isopenicillin N synthase"
FT /id="PRO_0000219503"
FT DOMAIN 176..288
FT /note="Fe2OG dioxygenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 87
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 87
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 91
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 91
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 183
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 183
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 189
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 189
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 211..216
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 214
FT /ligand="Fe(2+)"
FT /ligand_id="ChEBI:CHEBI:29033"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 214
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 216
FT /ligand="Fe(2+)"
FT /ligand_id="ChEBI:CHEBI:29033"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 216
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 270
FT /ligand="Fe(2+)"
FT /ligand_id="ChEBI:CHEBI:29033"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 279
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 281
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 281
FT /ligand="N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-
FT valine"
FT /ligand_id="ChEBI:CHEBI:58572"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT SITE 211
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT VARIANT 195
FT /note="I -> Y (in strain: AS-P-78)"
SQ SEQUENCE 331 AA; 37960 MW; E338FACBC7C74FC4 CRC64;
MASTPKANVP KIDVSPLFGD NMEEKMKVAR AIDAASRDTG FFYAVNHGVD VKRLSNKTRE
FHFSITDEEK WDLAIRAYNK EHQDQIRAGY YLSIPEKKAV ESFCYLNPNF KPDHPLIQSK
TPTHEVNVWP DEKKHPGFRE FAEQYYWDVF GLSSALLRGY ALALGKEEDF FSRHFKKEDA
LSSVVLIRYP YLNPIPPAAI KTAEDGTKLS FEWHEDVSLI TVLYQSDVAN LQVEMPQGYL
DIEADDNAYL VNCGSYMAHI TNNYYPAPIH RVKWVNEERQ SLPFFVNLGF NDTVQPWDPS
KEDGKTDQRP ISYGDYLQNG LVSLINKNGQ T
