AZAA_ASPNA
ID AZAA_ASPNA Reviewed; 2599 AA.
AC G3XMC4;
DT 02-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 14-DEC-2011, sequence version 1.
DT 03-AUG-2022, entry version 58.
DE RecName: Full=Non-reducing polyketide synthase azaA {ECO:0000303|PubMed:22921072};
DE Short=NR-PKS azaA {ECO:0000303|PubMed:22921072};
DE EC=2.3.1.- {ECO:0000269|PubMed:22921072};
DE AltName: Full=Azaphilone biosynthesis cluster protein azaA {ECO:0000303|PubMed:22921072};
GN Name=azaA {ECO:0000303|PubMed:22921072}; ORFNames=ASPNIDRAFT_56946;
OS Aspergillus niger (strain ATCC 1015 / CBS 113.46 / FGSC A1144 / LSHB Ac4 /
OS NCTC 3858a / NRRL 328 / USDA 3528.7).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=380704;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 1015 / CBS 113.46 / FGSC A1144 / LSHB Ac4 / NCTC 3858a / NRRL
RC 328 / USDA 3528.7;
RX PubMed=21543515; DOI=10.1101/gr.112169.110;
RA Andersen M.R., Salazar M.P., Schaap P.J., van de Vondervoort P.J.I.,
RA Culley D., Thykaer J., Frisvad J.C., Nielsen K.F., Albang R., Albermann K.,
RA Berka R.M., Braus G.H., Braus-Stromeyer S.A., Corrochano L.M., Dai Z.,
RA van Dijck P.W.M., Hofmann G., Lasure L.L., Magnuson J.K., Menke H.,
RA Meijer M., Meijer S.L., Nielsen J.B., Nielsen M.L., van Ooyen A.J.J.,
RA Pel H.J., Poulsen L., Samson R.A., Stam H., Tsang A., van den Brink J.M.,
RA Atkins A., Aerts A., Shapiro H., Pangilinan J., Salamov A., Lou Y.,
RA Lindquist E., Lucas S., Grimwood J., Grigoriev I.V., Kubicek C.P.,
RA Martinez D., van Peij N.N.M.E., Roubos J.A., Nielsen J., Baker S.E.;
RT "Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015
RT versus enzyme-producing CBS 513.88.";
RL Genome Res. 21:885-897(2011).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND INDUCTION.
RX PubMed=22921072; DOI=10.1016/j.chembiol.2012.07.004;
RA Zabala A.O., Xu W., Chooi Y.H., Tang Y.;
RT "Characterization of a silent azaphilone gene cluster from Aspergillus
RT niger ATCC 1015 reveals a hydroxylation-mediated pyran-ring formation.";
RL Chem. Biol. 19:1049-1059(2012).
CC -!- FUNCTION: Non-reducing polyketide synthase; part of the gene cluster
CC that mediates the biosynthesis of azaphilones, a class of fungal
CC metabolites characterized by a highly oxygenated pyrano-quinone
CC bicyclic core and exhibiting a broad range of bioactivities
CC (PubMed:22921072). In the first step, the non-reducing polyketide
CC synthase azaA forms the hexaketide precursor from successive
CC condensations of five malonyl-CoA units, presumably with a simple
CC acetyl-CoA starter unit (PubMed:22921072). The reactive polyketide
CC chain then undergoes a PT-mediated C2-C7 cyclization to afford the
CC aromatic ring and is eventually released as an aldehyde through the R-
CC domain (PubMed:22921072). The putative ketoreductase azaE is proposed
CC to catalyze the reduction of the terminal ketone resulting in the early
CC culture product FK17-P2a (PubMed:22921072). The monooxygenase azaH was
CC demonstrated to be the only enzyme required to convert FK17-P2a to
CC azanigerone E (PubMed:22921072). AzaH first hydroxylates the
CC benzaldehyde intermediate FK17-P2a at C4, which triggers the formation
CC of the pyran-ring to afford azanigerone E (PubMed:22921072). In
CC parallel, the 2,4-dimethylhexanoyl chain is synthesized by the HR-PKS
CC azaB and is proposed to be transferred to the C4-hydroxyl of
CC azanigerone E by the acyltransferase azaD directly from the ACP domain
CC of azaB (PubMed:22921072). Alternatively, the 2,4-dimethyl-hexanoyl
CC chain may be offloaded from the HR-PKS as a carboxylic acid and
CC converted to an acyl-CoA by azaF (PubMed:22921072). The resulting acyl-
CC CoA molecule could then be taken up as a substrate by AzaD to form
CC azanigerone B (PubMed:22921072). To yield the carboxylic acid
CC substituent in azanigerone A, the hydroxypropyl side chain of
CC azanigerone B would need to undergo a C-C oxidative cleavage catalyzed
CC by cytochrome P450 AzaI (PubMed:22921072). AzaI is proposed to act on a
CC vicinal diol that leads to a C-C bond scission either through an
CC alkoxyradical intermediate or a peroxy complex (PubMed:22921072). In
CC the biosynthesis of azanigerone A, azanigerone B first undergoes
CC hydroxylation at C10, possibly catalyzed by one of the two FAD-
CC dependent monooxygenases encoded in the cluster, azaG or azaL,
CC resulting in the vicinal diol azanigerone C (PubMed:22921072).
CC Oxidative cleavage of azanigerone C by azaI would yield the
CC corresponding aldehyde derivative of azanigerone A (PubMed:22921072).
CC Finally, the dehydrogenase azaJ is proposed to convert the aldehyde
CC functional group into the carboxylic acid, completing the conversion
CC from azanigerone B to azanigerone A (PubMed:22921072). Alternatively,
CC the oxidation of aldehyde to carboxylic acid may be catalyzed by the
CC same P450 enzyme azaI via consecutive oxidation or by endogenous
CC alcohol dehydrogenase (PubMed:22921072). {ECO:0000269|PubMed:22921072}.
CC -!- COFACTOR:
CC Name=pantetheine 4'-phosphate; Xref=ChEBI:CHEBI:47942;
CC Evidence={ECO:0000255};
CC Note=Binds 1 phosphopantetheine covalently. {ECO:0000255};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:22921072}.
CC -!- INDUCTION: Expression is under the control of the azaphilone cluster-
CC specific transcription factor azaR (PubMed:22921072).
CC {ECO:0000269|PubMed:22921072}.
CC -!- DOMAIN: Multidomain protein; including an N-terminal starter unit:ACP
CC transacylase (SAT) domain, a beta-ketoacyl synthase (KS) domain, a
CC malonyl-CoA:ACP transacylase (MAT) domain, a product template domain, a
CC acyl carrier protein (ACP) domain, a methyltransferase domain and a
CC reductive NADPH-binding domain that is required for NADPH-dependent
CC product release. {ECO:0000305|PubMed:22921072}.
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DR EMBL; ACJE01000001; EHA28237.1; -; Genomic_DNA.
DR AlphaFoldDB; G3XMC4; -.
DR SMR; G3XMC4; -.
DR PRIDE; G3XMC4; -.
DR EnsemblFungi; EHA28237; EHA28237; ASPNIDRAFT_56946.
DR VEuPathDB; FungiDB:ASPNIDRAFT2_1080089; -.
DR HOGENOM; CLU_000022_6_2_1; -.
DR Proteomes; UP000009038; Unassembled WGS sequence.
DR GO; GO:0016746; F:acyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0008168; F:methyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0031177; F:phosphopantetheine binding; IEA:InterPro.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR GO; GO:0044550; P:secondary metabolite biosynthetic process; IEA:UniProt.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.10.129.110; -; 1.
DR Gene3D; 3.40.366.10; -; 2.
DR Gene3D; 3.40.47.10; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR InterPro; IPR001227; Ac_transferase_dom_sf.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR014043; Acyl_transferase.
DR InterPro; IPR016035; Acyl_Trfase/lysoPLipase.
DR InterPro; IPR013120; Far_NAD-bd.
DR InterPro; IPR041068; HTH_51.
DR InterPro; IPR014031; Ketoacyl_synth_C.
DR InterPro; IPR014030; Ketoacyl_synth_N.
DR InterPro; IPR016036; Malonyl_transacylase_ACP-bd.
DR InterPro; IPR013217; Methyltransf_12.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR020841; PKS_Beta-ketoAc_synthase_dom.
DR InterPro; IPR042104; PKS_dehydratase_sf.
DR InterPro; IPR020806; PKS_PP-bd.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR006162; Ppantetheine_attach_site.
DR InterPro; IPR029063; SAM-dependent_MTases_sf.
DR InterPro; IPR032088; SAT.
DR InterPro; IPR016039; Thiolase-like.
DR Pfam; PF00698; Acyl_transf_1; 1.
DR Pfam; PF18558; HTH_51; 1.
DR Pfam; PF00109; ketoacyl-synt; 1.
DR Pfam; PF02801; Ketoacyl-synt_C; 1.
DR Pfam; PF08242; Methyltransf_12; 1.
DR Pfam; PF07993; NAD_binding_4; 1.
DR Pfam; PF00550; PP-binding; 1.
DR Pfam; PF16073; SAT; 1.
DR SMART; SM00827; PKS_AT; 1.
DR SMART; SM00825; PKS_KS; 1.
DR SMART; SM00823; PKS_PP; 1.
DR SUPFAM; SSF47336; SSF47336; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR SUPFAM; SSF52151; SSF52151; 1.
DR SUPFAM; SSF53335; SSF53335; 1.
DR SUPFAM; SSF53901; SSF53901; 1.
DR SUPFAM; SSF55048; SSF55048; 1.
DR PROSITE; PS50075; CARRIER; 1.
DR PROSITE; PS00012; PHOSPHOPANTETHEINE; 1.
PE 1: Evidence at protein level;
KW Acyltransferase; Methyltransferase; Multifunctional enzyme; NADP;
KW Phosphopantetheine; Phosphoprotein; Reference proteome; Transferase.
FT CHAIN 1..2599
FT /note="Non-reducing polyketide synthase azaA"
FT /id="PRO_0000437588"
FT DOMAIN 1653..1727
FT /note="Carrier"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 95..231
FT /note="N-terminal acylcarrier protein transacylase domain
FT (SAT)"
FT /evidence="ECO:0000250|UniProtKB:A0A0K0MCJ4"
FT REGION 375..793
FT /note="Ketosynthase (KS) domain"
FT /evidence="ECO:0000255"
FT REGION 902..1193
FT /note="Malonyl-CoA:ACP transacylase (MAT) domain"
FT /evidence="ECO:0000255"
FT REGION 1310..1589
FT /note="Product template (PT) domain"
FT /evidence="ECO:0000255"
FT REGION 1601..1652
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 1749..1779
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 1952..2140
FT /note="Methyltransferase domain"
FT /evidence="ECO:0000255"
FT REGION 2222..2467
FT /note="NADPH-binding (R) domain"
FT /evidence="ECO:0000255"
FT COMPBIAS 1601..1620
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1628..1652
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 1750..1779
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 132
FT /note="Nucleophile; for transacylase activity"
FT /evidence="ECO:0000250|UniProtKB:A0A0K0MCJ4"
FT ACT_SITE 250
FT /note="Proton donor/acceptor; for transacylase activity"
FT /evidence="ECO:0000250|UniProtKB:A0A0K0MCJ4"
FT ACT_SITE 539
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10022"
FT MOD_RES 1687
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
SQ SEQUENCE 2599 AA; 285933 MW; 020D14CBF0D7A79D CRC64;
MVTTTRATNP TNTLLLFGPQ ALSFSTATFA DIHARVVQTS ENAWIKQTIT SLPGLWDALV
KEFPQYGALE GKQLLRDLDR WFETGTMEHA EPHLPNILLS PMVVITQLTE YVDYLKTMPH
AADQQTETVG FCTGLLTALA ASLAPDIKGI RQYGAIAIKL AMIIGGVVDV QDITSPNGPS
KSLAVAWDSA ETQDRLNQII DQSPEAYISV EYDHNRATIT TAARSISSLQ QRLRNAGLIA
SQIGLRGRFH CACYEKDIEA LSKFCDSVPS LCLPDAAVLA LPTRSNDAGS FILSGKLHHC
ALRSILLDTS HWYQTLEVVR QSFLKSPSSM VVSFGPERCI PPSILKGLSS IVTTAAEYQP
SYLHRDPELC NPNEIAVIGM SCKVAGADDV DEFWDLLCKA ESQHQEVPKE RFGFESAFRE
VDPTRKWYGN FINEHDCFDH KFFKKSAREI AATDPQQRQM LQVAYQAVEQ SGYFTTPKSD
KDRKIGCYIG VCAADYEYNV ACHPPNAFMA TGNLKSFVAG KISHWFGWTG PGLCIDTACS
SSLVAVHQAC QAILTGDCTA ALAGGANIIT HPLWYQNLAA ASFLSPTGQC KPFDASADGY
CRGEGFAAVF LKKMSAAIAD GDMIIGSIKA TAVNQNQNCT PVFVPNAPTL SDLFRDVLDR
SQLTANQITV VEAHGTGTQV GDPAEYESIR NVLGGPSRST PLLFGSVKGL VGHTECTSGA
VSLVKTLLMQ QHEAIPPQPS FDRLNPEIPV SESDNMQIAT RFSPWTAEYR AALINNYGAC
GSNASMVVAQ APRTEQKRSA TRRTSVVLDY PFRLCGSDDR ALRAYSERLV RFIASGIKDG
ISVADLAFNV CRQSNPTLDR SLAFACRTTQ EVEEKLRAFV AGNQGLISAS RSKTPREVIL
CFGGQISNYV GLDREVYDNV ALLRKHLAIC DAACRDLGVD SIFPGIFQKS PISDPVKLQT
ILFSTQYSSA KAWMDSGVRP VAAVGHSFGE LTALCVTGIL SLADAMKMIV GRATVIRDFW
GEDKGSMIAV EADENRVQRL LAEAAKQCEL IHVRAPTIAC VNGPTSYTLA GPVKSIDIVT
EVISRLSDSG PSIRSKRLKV TNAFHSTLVE PLMEELEKVG QQLTFNAPTI QLERAIEHHS
DATLTSDYVP DHMRNPVYFN QAVQRLAQQY PDSVWLEAGS NSTITSMASR ALGSPKSLHF
QAVNITSDDS WSMLITSTLS LWKQGISTNF WAYHAKQTYE YNPVLLPPYQ FEPSRHWMEL
KVPSFMSNGN VQCGPRDEEE PPKTLWSLIE ASDKVARFQI NTAAPKYVEL VSGHVIANTA
PICPATVEVD IVVEALRSLR PDFMDSNLQP QVLAVTNQSP ICIDPNRSVW LECQAMDSNL
VWEWRIVSDS LQEPGTSSSA HVLGKLAFLS GQDEVKQQES EFMRLERLIG HQRCVDLLNT
TEADDIIQGR NIYTTFAGVV DYGEQYRGLK KIVGKGLESA GRVQKKPSEE SWLDAHLGDC
FSQVGGIWVN CMTDHNPDEM FIATGFEKWV RSPALRHGQP RPEIWDVLAC HHRSSEQTYL
TDIFIFDAEQ GALTEVILGI NYHKVAKASM SKILSRLSGT EAAPSSSTRA HPTSSSSPRL
PGPFVPEDKS QNETQTAGTN AVAKKKSEKS AQQNVLDKTR ALLAEISGLE PSEIEAETGL
ADIGIDSLMG MELARDLEAL FKCPLLGDEL ANVTTFQGLV EYVQSAVGVP ANGDEPDNTN
ADEVFEEDNL AASPSSSSSS TNLTEDSSLD PTETTTNISS YPELSPAWVL EAFEESKQLT
DHFIEQYRCA NYVDTILPKQ TQLCVALTVE AFEKLGCPIR SAVAGQKLER ILHIPKHAQL
AQYLYRLLSA DARLINLTED GRITRTHMAL PKPSDQILQD LLRLYPDHEW ANRLAAFTGA
RLAEVLKGET DGLGLIFGTD EGRELVAGLY GDSLLNKLSY RQMEDIITRL ASRIPRDSGP
LKILEMGAGT GGTTRGMAPL LARLGIPVEY TFTDLSGSFV AAARKKYQKE YPFMKFQVHD
IEKPPSDQLR HSQHIVIASN AIHATHSLTD SSRHVREFLK TDGFLMIVEM TQPVHWVDII
FGLFDGWWLF ADGRDHAIAS AGWWEKVFQS VGYGQVDWTD GHRPEVQIQR VIIALASGPR
YGRQPLPPAP PPNLVPGSHA SRQAAVNEYL DKYTKGFTLP AQTSNTDISN STSYWEKQCV
LITGATGSLG VHLVAAVAAL DDVQTVICLN RRSPMDPDLR QQQAFERRGI LLEAASMSKI
RVLQTDSSKP QLGLTDEVYS SLVTSTTHII HNAWPMTGKR PLSGLEQQFL VMRNLLDLAA
QCSSTRPANA PRIVFQFISS IATVGYYPLW SGQTLVPEAR MGIESVLANG YGEAKYVCEQ
MLDRTLHQYP DRFRAMAVRL GQIAGSRTSG YWNPMEHLSF LFKSAQTLQV FPDFTGDLCW
TPVNDVAATL SDLLLRSTHS NSTTDQPIYH IDNPVRQSWS EMVPVLIDAL GIPAQNVIPF
ADWVCRVRAF PGQVEWDNPS ALLIDFLDDH FLRMSCGGLL LDTKRACEHS PTLAAVGPVT
AELARKYIQS WKEMGFLNP
