IPNA_ACRCH
ID IPNA_ACRCH Reviewed; 338 AA.
AC P05189; Q00047;
DT 13-AUG-1987, integrated into UniProtKB/Swiss-Prot.
DT 13-AUG-1987, sequence version 1.
DT 03-AUG-2022, entry version 114.
DE RecName: Full=Isopenicillin N synthase {ECO:0000303|PubMed:3903520};
DE Short=IPNS {ECO:0000303|PubMed:3903520};
DE EC=1.21.3.1 {ECO:0000269|PubMed:3903520};
GN Name=PCBC; Synonyms=IPS;
OS Acremonium chrysogenum (Cephalosporium acremonium).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Hypocreales incertae sedis; Acremonium.
OX NCBI_TaxID=5044;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP PATHWAY.
RX PubMed=3903520; DOI=10.1038/318191a0;
RA Samson S.N., Belagaje R., Blankenship D.T., Chapman J.L., Perry D.,
RA Skatrud P.L., Vanfrank R.M., Abraham E.P., Baldwin J.E., Queener S.W.,
RA Ingolia T.D.;
RT "Isolation, sequence determination and expression in Escherichia coli of
RT the isopenicillin N synthetase gene from Cephalosporium acremonium.";
RL Nature 318:191-194(1985).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=N-2;
RX PubMed=2620834; DOI=10.1016/0378-1119(89)90493-9;
RA Ramsden M., McQuade B.A., Saunders K., Turner M.K., Harford S.;
RT "Characterization of a loss-of-function mutation in the isopenicillin N
RT synthetase gene of Acremonium chrysogenum.";
RL Gene 85:267-273(1989).
RN [3]
RP PROTEIN SEQUENCE OF 3-52, SUBUNIT, COFACTOR, FUNCTION, AND CATALYTIC
RP ACTIVITY.
RC STRAIN=ATCC 60777 / IMI 237183;
RX PubMed=3839755; DOI=10.1016/0014-5793(85)80382-3;
RA Baldwin J.E., Gagnon J., Ting H.H.;
RT "N-terminal amino acid sequence and some properties of isopenicillin-N
RT synthetase from Cephalosporium acremonium.";
RL FEBS Lett. 188:253-256(1985).
RN [4]
RP PROTEIN SEQUENCE OF 40-78 AND 237-264, AND PHOTOAFFINITY LABELLING.
RX PubMed=2317203;
RA Baldwin J.E., Coates J.B., Moloney M.G., Pratt A.J., Willis A.C.;
RT "Photoaffinity labelling of isopenicillin N synthetase.";
RL Biochem. J. 266:561-567(1990).
RN [5]
RP MUTAGENESIS OF HIS-272, FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8076799; DOI=10.1111/j.1574-6968.1994.tb07040.x;
RA Tiow-Suan S., Tan D.S.;
RT "Histidine-272 of isopenicillin N synthase of Cephalosporium acremonium,
RT which is possibly involved in iron binding, is essential for its catalytic
RT activity.";
RL FEMS Microbiol. Lett. 120:241-247(1994).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF HIS-49; HIS-64; HIS-116;
RP HIS-126; HIS-137; HIS-216 AND HIS-272.
RX PubMed=8557701; DOI=10.1074/jbc.271.2.889;
RA Tan D.S., Sim T.S.;
RT "Functional analysis of conserved histidine residues in Cephalosporium
RT acremonium isopenicillin N synthase by site-directed mutagenesis.";
RL J. Biol. Chem. 271:889-894(1996).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF ASP-218.
RX PubMed=9418249; DOI=10.1111/j.1574-6968.1997.tb12764.x;
RA Loke P., Sim J., Sim T.S.;
RT "Functional analysis of a conserved aspartate D218 in Cephalosporium
RT acremonium isopenicillin N synthase.";
RL FEMS Microbiol. Lett. 157:137-140(1997).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF GLN-234.
RX PubMed=9703965; DOI=10.1006/bbrc.1998.9016;
RA Loke P., Sim T.S.;
RT "Catalytic activity in Cephalosporium acremonium isopenicillin N synthase
RT does not involve glutamine-234.";
RL Biochem. Biophys. Res. Commun. 248:559-561(1998).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF GLN-227; GLN-234 AND
RP GLN-337.
RX PubMed=9826554; DOI=10.1006/bbrc.1998.9663;
RA Loke P., Sim T.S.;
RT "Analysis of glutamines in catalysis in Cephalosporium acremonium
RT isopenicillin N synthase by site-directed mutagenesis.";
RL Biochem. Biophys. Res. Commun. 252:472-475(1998).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF HIS-49; HIS-64; HIS-116;
RP HIS-126; HIS-137; HIS-216 AND HIS-272.
RX PubMed=9530516; DOI=10.1080/15216549800201352;
RA Tan D.S., Ang S.G., Sim T.S.;
RT "Involvement of a third histidine in the ferrous active site of
RT isopenicillin N synthase of Cephalosporium acremonium repudiated by
RT recombinant double histidine mutants.";
RL Biochem. Mol. Biol. Int. 44:333-345(1998).
RN [11]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF SER-283.
RX PubMed=9841222; DOI=10.1111/j.1574-6968.1998.tb13169.x;
RA Loke P., Sim T.S.;
RT "Mutational evidence for the role of serine-283 in Cephalosporium
RT acremonium isopenicillin N synthase.";
RL FEMS Microbiol. Lett. 165:353-356(1998).
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:3903520). IpnA catalyzes the cyclization of the
CC tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
CC (LLD-ACV or ACV) to form isopenicillin N (IPN) that contains the beta-
CC lactam nucleus (PubMed:3903520, PubMed:3839755, PubMed:8076799,
CC PubMed:8557701, PubMed:9418249, PubMed:9703965, PubMed:9826554,
CC PubMed:9530516, PubMed:9841222). The penicillin biosynthesis occurs via
CC 3 enzymatic steps, the first corresponding to the production of the
CC tripeptide N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
CC (LLD-ACV or ACV) by the NRPS pcbAB. The tripeptide ACV is then cyclized
CC to isopenicillin N (IPN) by the isopenicillin N synthase pcbC that
CC forms the beta-lactam nucleus. Finally, the alpha-aminoadipyl side
CC chain is exchanged for phenylacetic acid by the isopenicillin N
CC acyltransferase penDE to yield penicillin in the peroxisomal matrix (By
CC similarity). {ECO:0000250|UniProtKB:P08703, ECO:0000269|PubMed:3839755,
CC ECO:0000269|PubMed:3903520, ECO:0000269|PubMed:8076799,
CC ECO:0000269|PubMed:8557701, ECO:0000269|PubMed:9418249,
CC ECO:0000269|PubMed:9530516, ECO:0000269|PubMed:9703965,
CC ECO:0000269|PubMed:9826554, ECO:0000269|PubMed:9841222}.
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:3839755, ECO:0000269|PubMed:3903520,
CC ECO:0000269|PubMed:8076799, ECO:0000269|PubMed:8557701,
CC ECO:0000269|PubMed:9418249, ECO:0000269|PubMed:9530516,
CC ECO:0000269|PubMed:9703965, ECO:0000269|PubMed:9826554,
CC ECO:0000269|PubMed:9841222};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:22429;
CC Evidence={ECO:0000269|PubMed:3839755, ECO:0000269|PubMed:3903520,
CC ECO:0000269|PubMed:8076799, ECO:0000269|PubMed:8557701,
CC ECO:0000269|PubMed:9418249, ECO:0000269|PubMed:9530516,
CC ECO:0000269|PubMed:9703965, ECO:0000269|PubMed:9826554,
CC ECO:0000269|PubMed:9841222};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU00805,
CC ECO:0000269|PubMed:3839755};
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:3903520}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:3839755}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol
CC {ECO:0000250|UniProtKB:P08703}.
CC -!- SIMILARITY: Belongs to the iron/ascorbate-dependent oxidoreductase
CC family. {ECO:0000305}.
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DR EMBL; X03148; CAA26927.1; -; Genomic_DNA.
DR EMBL; M33522; AAA32674.1; -; Genomic_DNA.
DR PIR; S09312; S09312.
DR AlphaFoldDB; P05189; -.
DR SMR; P05189; -.
DR BioCyc; MetaCyc:MON-13364; -.
DR BRENDA; 1.21.3.1; 114.
DR SABIO-RK; P05189; -.
DR UniPathway; UPA00149; UER00240.
DR GO; GO:0005829; C:cytosol; ISS:GO_Central.
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; Direct protein sequencing; Iron;
KW Metal-binding; Oxidoreductase.
FT CHAIN 1..338
FT /note="Isopenicillin N synthase"
FT /id="PRO_0000219499"
FT DOMAIN 182..290
FT /note="Fe2OG dioxygenase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 89
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 89
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 93
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 93
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 185
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 185
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 191
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 191
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 213..218
FT /ligand="substrate"
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 218
FT /ligand="Fe(2+)"
FT /ligand_id="ChEBI:CHEBI:29033"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 218
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 272
FT /ligand="Fe(2+)"
FT /ligand_id="ChEBI:CHEBI:29033"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 281
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00805"
FT BINDING 283
FT /ligand="isopenicillin N"
FT /ligand_id="ChEBI:CHEBI:58399"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT BINDING 283
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 213
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:P05326"
FT SITE 283
FT /note="Involved in ACV-binding"
FT VARIANT 285
FT /note="P -> L (in strain: N-2; inactive)"
FT /evidence="ECO:0000269|PubMed:3903520"
FT MUTAGEN 49
FT /note="H->L: Reduces strongly the catalytic activiy."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 64
FT /note="H->L: Reduces the catalytic activiy."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 116
FT /note="H->L: Reduces strongly the catalytic activiy."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 126
FT /note="H->L: Reduces strongly the catalytic activiy."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 137
FT /note="H->L: Reduces the catalytic activiy."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 216
FT /note="H->L: Impairs the catalytic activity."
FT /evidence="ECO:0000269|PubMed:8557701,
FT ECO:0000269|PubMed:9530516"
FT MUTAGEN 218
FT /note="D->L: Impairs the catalytic activity."
FT /evidence="ECO:0000269|PubMed:9418249"
FT MUTAGEN 227
FT /note="Q->L: Does not affect the catalytic activity."
FT /evidence="ECO:0000269|PubMed:9826554"
FT MUTAGEN 234
FT /note="Q->L: Does not affect the catalytic activity."
FT /evidence="ECO:0000269|PubMed:9703965,
FT ECO:0000269|PubMed:9826554"
FT MUTAGEN 272
FT /note="H->L: Impairs the catalytic activity."
FT /evidence="ECO:0000269|PubMed:8076799,
FT ECO:0000269|PubMed:8557701, ECO:0000269|PubMed:9530516"
FT MUTAGEN 283
FT /note="S->A: Impairs the catalytic activity."
FT /evidence="ECO:0000269|PubMed:9841222"
FT MUTAGEN 337
FT /note="Q->L: Does not affect the catalytic activity."
FT /evidence="ECO:0000269|PubMed:9826554"
FT CONFLICT 22
FT /note="D -> T (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 177
FT /note="S -> F (in Ref. 2; AAA32674)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 338 AA; 38433 MW; CFB0B6E27FC04EE9 CRC64;
MGSVPVPVAN VPRIDVSPLF GDDKEKKLEV ARAIDAASRD TGFFYAVNHG VDLPWLSRET
NKFHMSITDE EKWQLAIRAY NKEHESQIRA GYYLPIPGKK AVESFCYLNP SFSPDHPRIK
EPTPMHEVNV WPDEAKHPGF RAFAEKYYWD VFGLSSAVLR GYALALGRDE DFFTRHSRRD
TTLSSVVLIR YPYLDPYPEP AIKTADDGTK LSFEWHEDVS LITVLYQSDV QNLQVKTPQG
WQDIQADDTG FLINCGSYMA HITDDYYPAP IHRVKWVNEE RQSLPFFVNL GWEDTIQPWD
PATAKDGAKD AAKDKPAISY GEYLQGGLRG LINKNGQT
