CCSB_ASPCL
ID CCSB_ASPCL Reviewed; 636 AA.
AC A1CLY7;
DT 07-JAN-2015, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 1.
DT 03-AUG-2022, entry version 62.
DE RecName: Full=Ketocytochalasin monooxygenase {ECO:0000305};
DE EC=1.14.13.- {ECO:0000269|PubMed:24838010};
DE AltName: Full=Carbonate-forming Baeyer-Villiger monooxygenase {ECO:0000303|PubMed:24838010};
DE Short=BVMO {ECO:0000303|PubMed:24838010};
DE AltName: Full=Cytochalasin biosynthesis protein B {ECO:0000303|PubMed:21983160};
GN Name=ccsB {ECO:0000303|PubMed:21983160}; ORFNames=ACLA_078650;
OS Aspergillus clavatus (strain ATCC 1007 / CBS 513.65 / DSM 816 / NCTC 3887 /
OS NRRL 1 / QM 1276 / 107).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Fumigati.
OX NCBI_TaxID=344612;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 1007 / CBS 513.65 / DSM 816 / NCTC 3887 / NRRL 1;
RX PubMed=18404212; DOI=10.1371/journal.pgen.1000046;
RA Fedorova N.D., Khaldi N., Joardar V.S., Maiti R., Amedeo P., Anderson M.J.,
RA Crabtree J., Silva J.C., Badger J.H., Albarraq A., Angiuoli S., Bussey H.,
RA Bowyer P., Cotty P.J., Dyer P.S., Egan A., Galens K., Fraser-Liggett C.M.,
RA Haas B.J., Inman J.M., Kent R., Lemieux S., Malavazi I., Orvis J.,
RA Roemer T., Ronning C.M., Sundaram J.P., Sutton G., Turner G., Venter J.C.,
RA White O.R., Whitty B.R., Youngman P., Wolfe K.H., Goldman G.H.,
RA Wortman J.R., Jiang B., Denning D.W., Nierman W.C.;
RT "Genomic islands in the pathogenic filamentous fungus Aspergillus
RT fumigatus.";
RL PLoS Genet. 4:E1000046-E1000046(2008).
RN [2]
RP FUNCTION, AND PATHWAY.
RX PubMed=21983160; DOI=10.1016/j.ymben.2011.09.008;
RA Qiao K., Chooi Y.H., Tang Y.;
RT "Identification and engineering of the cytochalasin gene cluster from
RT Aspergillus clavatus NRRL 1.";
RL Metab. Eng. 13:723-732(2011).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF ARG-421.
RC STRAIN=ATCC 1007 / CBS 513.65 / DSM 816 / NCTC 3887 / NRRL 1;
RX PubMed=24838010; DOI=10.1038/nchembio.1527;
RA Hu Y., Dietrich D., Xu W., Patel A., Thuss J.A., Wang J., Yin W.B.,
RA Qiao K., Houk K.N., Vederas J.C., Tang Y.;
RT "A carbonate-forming Baeyer-Villiger monooxygenase.";
RL Nat. Chem. Biol. 10:552-554(2014).
RN [4]
RP FUNCTION.
RX PubMed=27551732; DOI=10.1371/journal.pone.0161199;
RA Nielsen M.L., Isbrandt T., Petersen L.M., Mortensen U.H., Andersen M.R.,
RA Hoof J.B., Larsen T.O.;
RT "Linker flexibility facilitates module exchange in fungal hybrid PKS-NRPS
RT engineering.";
RL PLoS ONE 11:E0161199-E0161199(2016).
CC -!- FUNCTION: Ketocytochalasin monooxygenase; part of the gene cluster that
CC mediates the biosynthesis of a family of the mycotoxins cytochalasins E
CC and K (PubMed:21983160). The hybrid PKS-NRPS synthetase ccsA and the
CC enoyl reductase ccsC are responsible for fusion of phenylalanine with
CC an octaketide backbone and subsequent release of the stable tetramic
CC acid precursor (PubMed:21983160, PubMed:27551732). The polyketide
CC synthase module (PKS) of the PKS-NRPS ccsA is responsible for the
CC synthesis of the octaketide backbone (PubMed:21983160). The downstream
CC nonribosomal peptide synthetase (NRPS) amidates the carboxyl end of the
CC octaketide with a phenylalanine (PubMed:21983160). A reductase-like
CC domain (R) at the C-terminus catalyzes the reductive release of the
CC polyketide-amino acid intermediate (PubMed:21983160). Because ccsA
CC lacks a designated enoylreductase (ER) domain, the required activity is
CC provided the enoyl reductase ccsC (PubMed:21983160, PubMed:27551732).
CC Upon formation of the 11-membered carbocycle-fused perhydroisoindolone
CC intermediate, a number of oxidative steps are required to afford the
CC final cytochalasin E and K, including two hydroxylations at C17 and
CC C18, one alcohol oxidation at C17, one epoxidation at C6 and C7 and two
CC Baeyer-Villiger oxidations (PubMed:21983160). The oxidative
CC modification at C17, C18 and the C6-C7 epoxidation are likely to be
CC catalyzed by the two cytochrome P450 oxygenases ccsD and ccsG
CC (PubMed:21983160). CcsD may be responsible for the epoxidation of the
CC C6-C7 double bond (PubMed:21983160). CcsG may be responsible for the
CC successive oxidative modifications at C17 and C18 (PubMed:21983160).
CC The double Baeyer-Villiger oxidations of ketocytochalasin to
CC precytochalasin and cytochalasin Z(16) are among the final steps
CC leading to cytochalasin E and K and are catalyzed by ccsB
CC (PubMed:21983160, PubMed:24838010). The first oxygen insertion step
CC follows that of the classic BVMO mechanism, generating the ester
CC precytochalasin (PubMed:24838010). Release of precytochalasin into an
CC aqueous environment can generate the shunt product iso-precytochalasin
CC through spontaneous isomerization (PubMed:24838010). Alternatively,
CC precytochalasin can undergo further oxidation by ccsB to yield the in-
CC line carbonate-containing cytochalasin Z(16) (PubMed:24838010).
CC Cytochalasin Z(16) is a precursor to cytochalasin E and cytochalasin K,
CC whereas iso-precytochalasin is a precursor to cytochalasin Z(17) and
CC rosellichalasin (PubMed:21983160, PubMed:24838010). The hydrolyase ccsE
CC may catalyze hydrolysis of epoxide bond in cytochalasin E to afford
CC cytochalasin K (PubMed:21983160). The function of ccsF has not been
CC assigned but it may play a role in post-PKS-NRPS biosynthetic step,
CC resistance or transport of cytochalasins and related PKS-NRPS products
CC (PubMed:21983160). {ECO:0000269|PubMed:21983160,
CC ECO:0000269|PubMed:24838010, ECO:0000269|PubMed:27551732}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + ketocytochalasin + NADPH + O2 = H2O + iso-
CC precytochalasin + NADP(+); Xref=Rhea:RHEA:47700, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:87837, ChEBI:CHEBI:87838;
CC Evidence={ECO:0000269|PubMed:24838010};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + iso-precytochalasin + NADPH + O2 = cytochalasin Z16 +
CC H2O + NADP(+); Xref=Rhea:RHEA:47704, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:87838, ChEBI:CHEBI:87839;
CC Evidence={ECO:0000269|PubMed:24838010};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:24838010};
CC Note=Binds 1 FAD per subunit. {ECO:0000250|UniProtKB:Q47PU3};
CC -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000305|PubMed:21983160}.
CC -!- SIMILARITY: Belongs to the FAD-binding monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; DS027057; EAW09116.1; -; Genomic_DNA.
DR RefSeq; XP_001270542.1; XM_001270541.1.
DR AlphaFoldDB; A1CLY7; -.
DR SMR; A1CLY7; -.
DR EnsemblFungi; EAW09116; EAW09116; ACLA_078650.
DR GeneID; 4702675; -.
DR KEGG; act:ACLA_078650; -.
DR VEuPathDB; FungiDB:ACLA_078650; -.
DR eggNOG; KOG1399; Eukaryota.
DR HOGENOM; CLU_006937_8_2_1; -.
DR OMA; HIAYIIA; -.
DR OrthoDB; 405736at2759; -.
DR Proteomes; UP000006701; Unassembled WGS sequence.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW FAD; Flavoprotein; Monooxygenase; NADP; Oxidoreductase; Reference proteome.
FT CHAIN 1..636
FT /note="Ketocytochalasin monooxygenase"
FT /id="PRO_0000431478"
FT BINDING 125
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 133..136
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 143..145
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 145
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 151
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 195
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 279..285
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 302..303
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 420..421
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 534
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT SITE 421
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT MUTAGEN 421
FT /note="R->A: Completely abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:24838010"
SQ SEQUENCE 636 AA; 72109 MW; 60648ADF6C0C3480 CRC64;
MGSLLSIVLR SATRIIRPII PSSVLSLLSG PKPVHPLQTL QFDKEAILEK YQAERAKRLR
QDGVSQFKSA RSGAYDRFRQ DVARPQSREP IEAETKVLIV GAGFAGLVAA VKLDQQGVQD
FRIVDKAAGF GGTWYWNQYP GAACDVESLI YLPFLEETGY IPSRRFCYGP EIREQVNRVV
AKWDLARRSH LSTEITSMTW DESILRWHVK TNHGDHFITQ FVVMATGTFH EPKLPGIPGI
ENFKGDHFHS GRWDYRITGG DETGNMSQLA NKTVGIIGTG ASAVQLVPKL ARDAKKLYVF
QRTPSSIRFR DNSLYDTPSL KKSLPPGWQR QRMTDFANIL TGQVVDQDCD ALEGLQELTM
RAILKEASDA GVTVQPEQIP ELMQLADFRL MQQIRGQVDE IVKDQETAAK LKPWYSFMCK
RPTFHNDYLA AFNNPNVELV DTDGQGVSYL TETAVVANGR EYEVDLLVYS TGFDFDVEAN
FYRRTGIQLV GSRGRTFDEK WDEKGPSTLF GVHIREFPNL LYVGPAQTGV TANWTHTTYA
VGDHIAEFVA KSLRDGQYQA FEPTEEAEEE WGRRNEEGSE MRLLFAQSCP PGYYNREGKP
EEIPARWAYF PKGIIEWTRI TQEWREKGDY QGMDKR
