IPNA_PENRW
ID IPNA_PENRW Reviewed; 331 AA.
AC B6HLU0;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 16-DEC-2008, sequence version 1.
DT 03-AUG-2022, entry version 65.
DE RecName: Full=Isopenicillin N synthase {ECO:0000303|PubMed:1368505};
DE Short=IPNS {ECO:0000303|PubMed:1368505};
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:1368505}; Synonyms=pcbC;
GN ORFNames=PCH_Pc21g21380;
OS Penicillium rubens (strain ATCC 28089 / DSM 1075 / NRRL 1951 / Wisconsin
OS 54-1255) (Penicillium chrysogenum).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium;
OC Penicillium chrysogenum species complex.
OX NCBI_TaxID=500485;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 28089 / DSM 1075 / NRRL 1951 / Wisconsin 54-1255;
RX PubMed=18820685; DOI=10.1038/nbt.1498;
RA van den Berg M.A., Albang R., Albermann K., Badger J.H., Daran J.-M.,
RA Driessen A.J.M., Garcia-Estrada C., Fedorova N.D., Harris D.M.,
RA Heijne W.H.M., Joardar V.S., Kiel J.A.K.W., Kovalchuk A., Martin J.F.,
RA Nierman W.C., Nijland J.G., Pronk J.T., Roubos J.A., van der Klei I.J.,
RA van Peij N.N.M.E., Veenhuis M., von Doehren H., Wagner C., Wortman J.R.,
RA Bovenberg R.A.L.;
RT "Genome sequencing and analysis of the filamentous fungus Penicillium
RT chrysogenum.";
RL Nat. Biotechnol. 26:1161-1168(2008).
RN [2]
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 [3]
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 [4]
RP FUNCTION.
RX PubMed=2110531; DOI=10.1016/0014-5793(90)80224-7;
RA Whiteman P.A., Abraham E.P., Baldwin J.E., Fleming M.D., Schofield C.J.,
RA Sutherland J.D., Willis A.C.;
RT "Acyl coenzyme A: 6-aminopenicillanic acid acyltransferase from Penicillium
RT chrysogenum and Aspergillus nidulans.";
RL FEBS Lett. 262:342-344(1990).
RN [5]
RP FUNCTION.
RX PubMed=2120195; DOI=10.1128/jb.172.10.5908-5914.1990;
RA Tobin M.B., Fleming M.D., Skatrud P.L., Miller J.R.;
RT "Molecular characterization of the acyl-coenzyme A:isopenicillin N
RT acyltransferase gene (penDE) from Penicillium chrysogenum and Aspergillus
RT nidulans and activity of recombinant enzyme in Escherichia coli.";
RL J. Bacteriol. 172:5908-5914(1990).
RN [6]
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 [7]
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 [8]
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 [9]
RP FUNCTION.
RX PubMed=8416970; DOI=10.1016/s0021-9258(18)54203-4;
RA Lendenfeld T., Ghali D., Wolschek M., Kubicek-Pranz E.M., Kubicek C.P.;
RT "Subcellular compartmentation of penicillin biosynthesis in Penicillium
RT chrysogenum. The amino acid precursors are derived from the vacuole.";
RL J. Biol. Chem. 268:665-671(1993).
RN [10]
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 [11]
RP FUNCTION.
RX PubMed=9266851; DOI=10.1006/bbrc.1997.7107;
RA Etchegaray A., Dieckmann R., Kennedy J., Turner G., von Doehren H.;
RT "ACV synthetase: expression of amino acid activating domains of the
RT Penicillium chrysogenum enzyme in Aspergillus nidulans.";
RL Biochem. Biophys. Res. Commun. 237:166-169(1997).
RN [12]
RP FUNCTION.
RX PubMed=9355751; DOI=10.1042/bj3270185;
RA Theilgaard H.B., Kristiansen K.N., Henriksen C.M., Nielsen J.;
RT "Purification and characterization of delta-(L-alpha-aminoadipyl)-L-
RT cysteinyl-D-valine synthetase from Penicillium chrysogenum.";
RL Biochem. J. 327:185-191(1997).
RN [13]
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 [14]
RP FUNCTION.
RX PubMed=19686863; DOI=10.1016/j.ymben.2009.08.002;
RA Siewers V., Chen X., Huang L., Zhang J., Nielsen J.;
RT "Heterologous production of non-ribosomal peptide LLD-ACV in Saccharomyces
RT cerevisiae.";
RL Metab. Eng. 11:391-397(2009).
RN [15]
RP FUNCTION.
RX PubMed=21889568; DOI=10.1016/j.biochi.2011.08.002;
RA Wu X., Garcia-Estrada C., Vaca I., Martin J.F.;
RT "Motifs in the C-terminal region of the Penicillium chrysogenum ACV
RT synthetase are essential for valine epimerization and processivity of
RT tripeptide formation.";
RL Biochimie 94:354-364(2012).
RN [16]
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).
RN [17]
RP FUNCTION.
RX PubMed=22777282; DOI=10.1007/s00253-012-4256-0;
RA Fernandez-Aguado M., Teijeira F., Martin J.F., Ullan R.V.;
RT "A vacuolar membrane protein affects drastically the biosynthesis of the
RT ACV tripeptide and the beta-lactam pathway of Penicillium chrysogenum.";
RL Appl. Microbiol. Biotechnol. 97:795-808(2013).
RN [18]
RP FUNCTION.
RX PubMed=23053082; DOI=10.1007/s00253-012-4425-1;
RA Fernandez-Aguado M., Ullan R.V., Teijeira F., Rodriguez-Castro R.,
RA Martin J.F.;
RT "The transport of phenylacetic acid across the peroxisomal membrane is
RT mediated by the PaaT protein in Penicillium chrysogenum.";
RL Appl. Microbiol. Biotechnol. 97:3073-3084(2013).
RN [19]
RP FUNCTION.
RX PubMed=24480587; DOI=10.1016/j.ymben.2014.01.004;
RA Fernandez-Aguado M., Martin J.F., Rodriguez-Castro R., Garcia-Estrada C.,
RA Albillos S.M., Teijeira F., Ullan R.V.;
RT "New insights into the isopenicillin N transport in Penicillium
RT chrysogenum.";
RL Metab. Eng. 22:89-103(2014).
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). The first step of the pathway is performed
CC by the trimodular NRPS acvA that produces the tripeptide N-[(5S)-5-
CC amino-5-carboxypentanoyl]-L-cysteinyl-D-valine (LLD-ACV or ACV) via
CC condensation of the 3 residues L-2-aminoadipate, L-cysteine and L-
CC valine (PubMed:9266851, PubMed:9355751, PubMed:19686863,
CC PubMed:21889568). The precursor amino acids for penicillin biosynthesis
CC are withdrawn from the vacuolar amino acid pool by the MFS-type
CC transporter penV (PubMed:8416970, PubMed:22777282). Each of the
CC constituent amino acids of the tripeptide acv are activated as
CC aminoacyl-adenylates with peptide bonds formed through the
CC participation of amino acid thioester intermediates (PubMed:9266851,
CC PubMed:21889568). The tripeptide ACV is then cyclized to form
CC isopenicillin N (IPN) by the isopenicillin N synthase ipnA that forms
CC the beta-lactam nucleus (PubMed:1368505, PubMed:1369045,
CC PubMed:1588566). Finally, the alpha-aminoadipyl side chain is exchanged
CC for phenylacetic acid by the isopenicillin N acyltransferase aatA to
CC yield penicillin (PubMed:2120195, PubMed:2110531, PubMed:1368505). This
CC step occurs in the peroxisomal matrix and the penM and paaT
CC transporters are involved in the isopenicillin N and phenylacetic acid
CC import into the peroxisome, respectively (PubMed:23053082,
CC PubMed:24480587). {ECO:0000269|PubMed:1368505,
CC ECO:0000269|PubMed:1369045, ECO:0000269|PubMed:1588566,
CC ECO:0000269|PubMed:19686863, ECO:0000269|PubMed:2110531,
CC ECO:0000269|PubMed:2120195, ECO:0000269|PubMed:21889568,
CC ECO:0000269|PubMed:22777282, ECO:0000269|PubMed:23053082,
CC ECO:0000269|PubMed:24480587, ECO:0000269|PubMed:8416970,
CC ECO:0000269|PubMed:9266851, ECO:0000269|PubMed:9355751}.
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 -!- 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; AM920436; CAP97035.1; -; Genomic_DNA.
DR RefSeq; XP_002569113.1; XM_002569067.1.
DR SMR; B6HLU0; -.
DR STRING; 1108849.XP_002569113.1; -.
DR EnsemblFungi; CAP97035; CAP97035; PCH_Pc21g21380.
DR GeneID; 8310374; -.
DR KEGG; pcs:Pc21g21380; -.
DR VEuPathDB; FungiDB:PCH_Pc21g21380; -.
DR eggNOG; KOG0143; Eukaryota.
DR HOGENOM; CLU_010119_6_1_1; -.
DR OMA; EADDNAY; -.
DR OrthoDB; 755305at2759; -.
DR UniPathway; UPA00149; UER00240.
DR Proteomes; UP000000724; Contig Pc00c21.
DR GO; GO:0005829; C:cytosol; IEA:UniProtKB-SubCell.
DR GO; GO:0051213; F:dioxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
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; Reference proteome.
FT CHAIN 1..331
FT /note="Isopenicillin N synthase"
FT /id="PRO_0000455268"
FT DOMAIN 181..288
FT /note="Fe2OG dioxygenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 214
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 216
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 270
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
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"
SQ SEQUENCE 331 AA; 38010 MW; EADFEE9BC7C75B94 CRC64;
MASTPKANVP KIDVSPLFGD NMEEKMKVAR AIDAASRDTG FFYAVNHGVD VKRLSNKTRE
FHFSITDEEK WDLAIRAYNK EHQDQIRAGY YLSIPEKKAV ESFCYLNPNF KPDHPLIQSK
TPTHEVNVWP DEKKHPGFRE FAEQYYWDVF GLSSALLRGY ALALGKEEDF FSRHFKKEDA
LSSVVLIRYP YLNPYPPAAI KTAEDGTKLS FEWHEDVSLI TVLYQSDVAN LQVEMPQGYL
DIEADDNAYL VNCGSYMAHI TNNYYPAPIH RVKWVNEERQ SLPFFVNLGF NDTVQPWDPS
KEDGKTDQRP ISYGDYLQNG LVSLINKNGQ T
