BSLS_BEABA
ID BSLS_BEABA Reviewed; 3146 AA.
AC D1FVF0;
DT 11-DEC-2019, integrated into UniProtKB/Swiss-Prot.
DT 19-JAN-2010, sequence version 1.
DT 03-AUG-2022, entry version 61.
DE RecName: Full=Bassianolide nonribosomal cyclodepsipeptide synthetase {ECO:0000303|PubMed:19285149};
DE Short=BSLS {ECO:0000303|PubMed:19285149};
DE Includes:
DE RecName: Full=Nonribosomal peptide synthetase {ECO:0000303|PubMed:19285149};
DE EC=6.1.2.- {ECO:0000269|PubMed:23608474, ECO:0000269|PubMed:23727842, ECO:0000269|PubMed:29163920, ECO:0000269|PubMed:31388353, ECO:0000269|PubMed:31471217};
DE Includes:
DE RecName: Full=S-adenosyl-L-methionine-dependent N-methyltransferase {ECO:0000303|PubMed:19285149};
DE EC=2.1.1.- {ECO:0000269|PubMed:29163920, ECO:0000269|PubMed:31388353, ECO:0000269|PubMed:31471217};
GN Name=BSLS {ECO:0000303|PubMed:19285149};
OS Beauveria bassiana (White muscardine disease fungus) (Tritirachium
OS shiotae).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Cordycipitaceae; Beauveria.
OX NCBI_TaxID=176275;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, DISRUPTION PHENOTYPE, DOMAIN,
RP AND BIOTECHNOLOGY.
RC STRAIN=ATCC 7159;
RX PubMed=19285149; DOI=10.1016/j.fgb.2009.03.001;
RA Xu Y., Orozco R., Kithsiri Wijeratne E.M., Espinosa-Artiles P.,
RA Leslie Gunatilaka A.A., Patricia Stock S., Molnar I.;
RT "Biosynthesis of the cyclooligomer depsipeptide bassianolide, an
RT insecticidal virulence factor of Beauveria bassiana.";
RL Fungal Genet. Biol. 46:353-364(2009).
RN [2]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=23608474; DOI=10.1016/j.ymben.2013.04.001;
RA Yu D., Xu F., Zi J., Wang S., Gage D., Zeng J., Zhan J.;
RT "Engineered production of fungal anticancer cyclooligomer depsipeptides in
RT Saccharomyces cerevisiae.";
RL Metab. Eng. 18:60-68(2013).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND DOMAIN.
RX PubMed=23727842; DOI=10.1039/c3cc42425a;
RA Yu D., Xu F., Gage D., Zhan J.;
RT "Functional dissection and module swapping of fungal cyclooligomer
RT depsipeptide synthetases.";
RL Chem. Commun. (Camb.) 49:6176-6178(2013).
RN [4]
RP INDUCTION.
RX PubMed=25912088; DOI=10.1016/j.jip.2015.04.004;
RA Lobo L.S., Luz C., Fernandes E.K., Juarez M.P., Pedrini N.;
RT "Assessing gene expression during pathogenesis: Use of qRT-PCR to follow
RT toxin production in the entomopathogenic fungus Beauveria bassiana during
RT infection and immune response of the insect host Triatoma infestans.";
RL J. Invertebr. Pathol. 128:14-21(2015).
RN [5]
RP BIOTECHNOLOGY.
RX PubMed=27676608; DOI=10.1016/j.bioorg.2016.09.008;
RA Mun B., Park Y.J., Sung G.H., Lee Y., Kim K.H.;
RT "Synthesis and antitumor activity of (-)-bassianolide in MDA-MB 231 breast
RT cancer cells through cell cycle arrest.";
RL Bioorg. Chem. 69:64-70(2016).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, AND DOMAIN.
RX PubMed=29163920; DOI=10.1039/c7sc03093b;
RA Steiniger C., Hoffmann S., Mainz A., Kaiser M., Voigt K., Meyer V.,
RA Suessmuth R.D.;
RT "Harnessing fungal nonribosomal cyclodepsipeptide synthetases for
RT mechanistic insights and tailored engineering.";
RL Chem. Sci. 8:7834-7843(2017).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, AND DOMAIN.
RX PubMed=31471217; DOI=10.1016/j.chembiol.2019.08.005;
RA Steiniger C., Hoffmann S., Suessmuth R.D.;
RT "Probing Exchange Units for Combining Iterative and Linear Fungal
RT Nonribosomal Peptide Synthetases.";
RL Cell Chem. Biol. 0:0-0(2019).
RN [8]
RP FUNCTION, CATALYTIC ACTIVITY, DOMAIN, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=31388353; DOI=10.1186/s13036-019-0195-y;
RA Xu F., Butler R., May K., Rexhepaj M., Yu D., Zi J., Chen Y., Liang Y.,
RA Zeng J., Hevel J., Zhan J.;
RT "Modified substrate specificity of a methyltransferase domain by protein
RT insertion into an adenylation domain of the bassianolide synthetase.";
RL J. Biol. Eng. 13:65-65(2019).
CC -!- FUNCTION: Bassianolide nonribosomal synthetase that mediates the
CC biosynthesis of bassianolide (BSL), a non-ribosomal cyclodepsipeptide
CC that shows insecticidal and cancer cell antiproliferative activity
CC (PubMed:19285149, PubMed:23608474, PubMed:23727842, PubMed:29163920,
CC PubMed:31471217, PubMed:31388353). BSLS first catalyzes the iterative
CC synthesis of an enzyme-bound dipeptidol monomer intermediate from D-2-
CC hydroxyisovalerate and L-leucine before performing the condensation and
CC cyclization of 4 dipeptidol monomers to yield the cyclic tetrameric
CC ester bassianolide (PubMed:23608474, PubMed:23727842, PubMed:29163920,
CC PubMed:31471217, PubMed:31388353). The N-methyltransferase MT domain is
CC responsible for the methylation of the leucine residues of bassianolide
CC (PubMed:29163920, PubMed:31388353). BSLS is flexible with both the
CC amino acid and hydroxyl acid precursors, and produces bassianolide as
CC the major product (containing N-methyl-L-Leu), together with small
CC amounts of beauvericin and its analogs beauvericins A-C (containing N-
CC methyl-L-Phe) (PubMed:31388353). {ECO:0000269|PubMed:19285149,
CC ECO:0000269|PubMed:23608474, ECO:0000269|PubMed:23727842,
CC ECO:0000269|PubMed:29163920, ECO:0000269|PubMed:31388353,
CC ECO:0000269|PubMed:31471217}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4 (R)-2-hydroxy-3-methylbutyrate + 8 ATP + 4 L-leucine + 4 S-
CC adenosyl-L-methionine = 8 AMP + bassianolide + 8 diphosphate + 8 H(+)
CC + 4 S-adenosyl-L-homocysteine; Xref=Rhea:RHEA:62272,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:57427, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789,
CC ChEBI:CHEBI:145108, ChEBI:CHEBI:145660, ChEBI:CHEBI:456215;
CC Evidence={ECO:0000269|PubMed:23608474, ECO:0000269|PubMed:23727842,
CC ECO:0000269|PubMed:29163920, ECO:0000269|PubMed:31388353,
CC ECO:0000269|PubMed:31471217};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62273;
CC Evidence={ECO:0000269|PubMed:23608474, ECO:0000269|PubMed:23727842,
CC ECO:0000269|PubMed:29163920, ECO:0000269|PubMed:31388353,
CC ECO:0000269|PubMed:31471217};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.46 uM for L-phenylalanyl-N-acetyl-cysteamine thioester (by the
CC MT domain) {ECO:0000269|PubMed:31388353};
CC KM=2.8 uM for L-leucyl-N-acetyl-cysteamine thioester(by the MT
CC domain) {ECO:0000269|PubMed:31388353};
CC Vmax=0.41 uM/min/mg enzyme toward L-phenylalanyl-N-acetyl-cysteamine
CC thioester (by the MT domain) {ECO:0000269|PubMed:31388353};
CC Vmax=0.05 uM/min/mg enzyme toward L-leucyl-N-acetyl-cysteamine
CC thioester (by the MT domain) {ECO:0000269|PubMed:31388353};
CC -!- INDUCTION: Expression is induced during insect infection.
CC {ECO:0000269|PubMed:25912088}.
CC -!- DOMAIN: NRP synthetases are composed of discrete domains (adenylation
CC (A), thiolation (T) or peptidyl carrier protein (PCP) and condensation
CC (C) domains) which when grouped together are referred to as a single
CC module. Each module is responsible for the recognition (via the A
CC domain) and incorporation of a single amino acid into the growing
CC peptide product. Thus, an NRP synthetase is generally composed of one
CC or more modules and can terminate in a thioesterase domain (TE) that
CC releases the newly synthesized peptide from the enzyme. Occasionally,
CC additional domains required for further modifications are also present
CC (Probable). Bassianolide synthetase has the C1-A1-T1-C2-A2-MT-T2a-T2b-
CC C3 domain organization. During catalysis, C3 and C2 take turns to
CC incorporate the two biosynthetic precursors into the growing
CC depsipeptide chain that swings between T1 and T2a/T2b with C3 cyclizing
CC the chain when it reaches the full length (PubMed:23727842,
CC PubMed:29163920, PubMed:31471217, PubMed:31388353). The N-
CC methyltransferase MT domain in module 2 is responsible for the
CC methylation of the leucine residues integrated in the backbone
CC structure (PubMed:29163920, PubMed:31388353).
CC {ECO:0000269|PubMed:23727842, ECO:0000269|PubMed:29163920,
CC ECO:0000269|PubMed:31388353, ECO:0000269|PubMed:31471217, ECO:0000305}.
CC -!- DISRUPTION PHENOTYPE: Abolishes the production of bassianolide but does
CC not affect the biosynthesis of beauvericin (PubMed:19285149).
CC Attenuates the virulence against insects, including the corn earworm
CC (Helicoverpa zea), the fall armyworm (Spodoptera exigua) and the
CC greater wax moth (Galleria mellonella) (PubMed:19285149).
CC {ECO:0000269|PubMed:19285149}.
CC -!- BIOTECHNOLOGY: Shows insecticidal activity and contributes to the
CC virulence of the fungus against insects including the corn earworm
CC (Helicoverpa zea), the fall armyworm (Spodoptera exigua) and the
CC greater wax moth (Galleria mellonella) (PubMed:19285149). Bassianolide
CC possesses antitumor activities and displays significant cytotoxicity
CC against several human tumor cell lines, including A549, SK-OV-3, HepG2,
CC HCT-15, MCF-7 and MDA-MB 231 cell lines with IC(50) values of 7.24,
CC 8.44, 15.39, 6.40, 11.42 and 3.98 ug/ml respectively (PubMed:27676608).
CC Bassianolide especially induces G0/G1 arrest associated with a decrease
CC of cyclin A, D1 and an increase of p53, MDM2, and p21 expression in
CC human breast adenocarcinoma cell lines (PubMed:27676608).
CC {ECO:0000269|PubMed:19285149, ECO:0000269|PubMed:27676608}.
CC -!- SIMILARITY: Belongs to the NRP synthetase family. {ECO:0000305}.
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DR EMBL; FJ439897; ACR78148.1; -; Genomic_DNA.
DR SMR; D1FVF0; -.
DR STRING; 176275.XP_008595949.1; -.
DR GO; GO:0016853; F:isomerase activity; IEA:UniProtKB-KW.
DR GO; GO:0016874; F:ligase 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:0043604; P:amide biosynthetic process; IEA:UniProt.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR GO; GO:1901566; P:organonitrogen compound biosynthetic process; IEA:UniProt.
DR Gene3D; 1.10.1200.10; -; 2.
DR Gene3D; 3.30.300.30; -; 3.
DR Gene3D; 3.30.559.10; -; 3.
DR Gene3D; 3.40.50.12780; -; 1.
DR Gene3D; 3.40.50.150; -; 1.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR010071; AA_adenyl_domain.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR045851; AMP-bd_C_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR023213; CAT-like_dom_sf.
DR InterPro; IPR001242; Condensatn.
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 Pfam; PF00501; AMP-binding; 2.
DR Pfam; PF00668; Condensation; 2.
DR Pfam; PF00550; PP-binding; 3.
DR SMART; SM00823; PKS_PP; 3.
DR SUPFAM; SSF47336; SSF47336; 3.
DR SUPFAM; SSF53335; SSF53335; 1.
DR TIGRFAMs; TIGR01733; AA-adenyl-dom; 2.
DR PROSITE; PS00455; AMP_BINDING; 2.
DR PROSITE; PS50075; CARRIER; 3.
DR PROSITE; PS00012; PHOSPHOPANTETHEINE; 3.
PE 1: Evidence at protein level;
KW Isomerase; Ligase; Methyltransferase; Multifunctional enzyme;
KW Phosphopantetheine; Phosphoprotein; Repeat; S-adenosyl-L-methionine;
KW Transferase.
FT CHAIN 1..3146
FT /note="Bassianolide nonribosomal cyclodepsipeptide
FT synthetase"
FT /id="PRO_0000448666"
FT DOMAIN 1015..1091
FT /note="Carrier 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT DOMAIN 2514..2588
FT /note="Carrier 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT DOMAIN 2614..2688
FT /note="Carrier 3"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT REGION 1..23
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 69..454
FT /note="Condensation 1"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:19285149"
FT REGION 495..887
FT /note="Adenylation 1"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:19285149"
FT REGION 1109..1538
FT /note="Condensation 2"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:19285149"
FT REGION 1567..1973
FT /note="Adenylation 2"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:19285149"
FT REGION 2041..2181
FT /note="S-adenosyl-L-methionine-dependent N-
FT methyltransferase (MT)"
FT /evidence="ECO:0000255, ECO:0000269|PubMed:31388353"
FT REGION 2734..3138
FT /note="Condensation 3"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:19285149"
FT COMPBIAS 1..22
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOD_RES 1052
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MOD_RES 2548
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
FT MOD_RES 2648
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
SQ SEQUENCE 3146 AA; 348272 MW; C6E7E94E0CE6B55A CRC64;
MEPPNNANTG QLGPTLPNGT VDLPTDLSRE ITRHFGLEQD EIEEILPCTP FQRDVIECAS
DDKRRAVGHV VYEIPEDVDT ERLAAAWKAT VRYTPALRTC IFTSETGNAF QVVLRDYFIF
ARMYCPSAHL KSAIVKDEAT AAVAGPRCNR YVLTGEPNSK RRVLVWTFSH SFVDSAFQGR
ILQQVLAAYK DGHGRVFSLQ PTTDLTESEN GDHLSTPASE RTVVIERATQ FWQEKLHGLD
ASVFPHLPSH KRVPAIDARA DHYLPCPPFI QHEWSSTTVC RTALAILLAR YTHSSEALFG
VVTEQSHEEH PLLLDGPTST VVPFRVLCAL NQSVSKVMEA ITTYDHDMRQ FAHAGLCNIS
RIGDDASAAC GFQTVLMVTD SRTAGDDEIH QVLEESEKFI PCTDRALLLS CQMTDEGVLL
VARYDQSILE PLQMARFLRQ LGFLINKLQS TDGSPCVGQL DVLAPEDRTE IEGWNSEPLQ
TQDCLIHSEV VRNAGDTPNK PAVCAWDGEW TYSELNNVSS RLASYISSLD LGQQLIVPIY
LEKSKWVMAA ILAVLKAGHA FTLIDPNDPP ARTAQIIKQA SASIALTSAL HQSKMQAVVG
RCITVDDDLV QTLTTFEGSQ VASAAKPGDL AYVIFTSGST GDPKGIMIEH RAFYSSVVKF
GKALGIRSST RALQFATHGF GAFLLEVLTT LIHGGCICVP SDHDRMHNIP GFIRQNQINW
MMATPSYMTT MKPEDVPGLE TLVLVGEQMS SSINDVWLSE LQLLDGYGQS ESSSICFVGK
IDDSSRDPNN LGWAIGAHSW IINPDNPDQL VPIGAIGELL IESPGIARGY LFSQSTETPF
LERAPAWYAS KQPPYGVKFY RTGDLARYAP DGTVICLGRM DSQVKIRGQR VELDAIENLL
RRQFPSDVTV VAEAVKRSDL PSSVVITGFL ISSEYVVGAP STEDTYILDQ VVTQEINAKM
RQILPAHSIP SFYICMKSLP RTATGKVDRR KLRSIGSSLL ALQAQSTAPR SSQAPDASAG
VTKLEEVWMD IFNLTPNSHN IGGNFFALGG DSITAIKMVN MARAAGIQLK VSDIFQNPTL
ASLQAAIGGS SMTVTSIPAL ALDGPVEQSY SQGRLWFLDQ LEIGANWYTI PYAVRLRGPL
DVDALNRALL ALEKRHETLR TTFEDQDGVG VQIIHETLLD QLRIINADHA DYVQLLKQEQ
TAPFNLASES GWRVSLIRLD DDDNILSIVM HHIISDGWSI DVLRRELGQL YAAALHGADL
FGSALSPLPI QYRDFSVWQK QDAQVAEHER QLQYWQKQLA DCSPAKLPTD FHRPALLSGK
ATTVPVTITS ELYYRLQEFC STFNTTSFVV LLATFRAAHY RLTGVDDAVI GTPIANRNRH
ELENLIGFFV NTQCMRITIN EDEDTFESLV RQVRSTTTAA FEHEDVPFER VVSAMLPGSR
DLSQNPLAQL VFAIHSHKDL GKFELEALES EPLQNEVYTR FDAEFHFFQA PDGLTGYINF
ATELFKVETI QNVVSVFLQI LRHGLEHPQT LISVVPLTDG LAELRSMGLL EIKKVEYPRD
SSVVDVFRTQ VASYPDTLAV VDSSSRLTYA ELDHQSDLLA TWLRQQNLPT EALVVVLAPR
SCETIITFLG ILKANLAYLP LDIRSPITRM RDVLSTLPGR TIALLCSDEV APDFQLPSIE
LVRIADALEE AAGMTSLNGH EHVPVPSPSP TSLAYVLYTS GSTGRPKGVM IEHRAIVRLA
RSDIIPDYRP ACGDTMAHMF NTAFDGATYE IYTMLLNGGT LVCVDYMDTL SPKSLEAVFK
KEQVNATIMA PALLKLYLAD ARDALKGLDV LISGGDRFDP QDAVDAQSLV RGSCYNGYGP
TENGVFSTVY KVDKNDPFVN GVPLGRAVNN SGAYVVDRNQ QLVGPGIIGE LVVTGDGLAR
GYTERAFDQN RFTQLKVEGQ SVRGYRTGDR VRYRVGEGLI EFFGRMDFQF KIRSNRIEAG
EVEAAILSHP AVRNAAVILR VEEKLEPEIV GFVVAEHDDT AEQEEAGDQV EGWQAFFEST
TYTELDTVSS SEIGKDFKGW TSMYDGNEID KAEMQEWLDD TIHTLTDGQA LGHVLEIGTG
SGMVLFNLGS GLQSFVGLEP SKSAAAFVNN AIKSTPALAG KAQVFVGTAT DTNKLDDLHP
DLVIFNSVLQ YFPTRDYLER VVDALVHLRS AKRIFFGDVR SYATNRHFLA ARAIYTLGNH
TTKDEVRKKM AEMEEREEEF LVEPAFFTTL VNRLPDVRHV EIIPKNMQAT NELSAYRYAA
VVHLRGSDEL TRPVHPIKMD DWVDFQASHM HKDALREYLR LAENTKTVAI SNIPYGKTIF
ERQVVESLDE TSEDAPHASL DGAAWISAVR SDAKARSSLS VPDLVLLAKE TGFRVEVSAA
RQWSQSGALD AVFHRYPAEP GVRTLFQFPT DNDVRMSAPL TNQPLQRLQK RRVAVQVREW
LQDRIPSYMI PSHIVALDQM PLNTSGKVDR KELSRQAKAI KKVQKSAPPT APAFPLSEVE
VMLCEELTKT FEMDVNITDD FFQLGGHSLL ATRLVARISH RLGARLTVKD VFDYPVFSEL
ADIIRQQLAS KNTLLPTASA GGGGQDKKES AGVAPTTDME AMLCEEFANI LGMDVGITDN
FFDLGGHSLM ATRLAARIGH RLNTTISVKD IFSHPVIFQL SAKLEVSQLE SSSGGTDIKM
PDYTAFQLIP AADAEKFMQD HIYPQINFSQ DMVQDVYLAT HLQQCFLRDV FGRPKPLVPF
YVEFPPDSNP HTLATACTSL VDKYDIFRTI FVEAEGNLYQ VVLKHLNLDI DVVETDANVH
KTSSDLVDAI AKEPVRLGQP MIQVKVLKQT SSVRVLLWLS HALYDGLSWE HIVRDLHILS
KERSLPPATQ FSRYMQYVDH TRGPGCDFWR DVLQNAPITN LSDAGSGGRP TKAGDPRVWH
AGKVISGPSQ AIRSSITQAT VFNAACAIVL SKETGTDNVV FGRIVSGRQG LPVRWQNIIG
PCTNAVPVRA VVDAHGNHQQ MLRDLQEQYL LSLPYETIGF DEIKRSCTDW PDSARNYGCC
VTYQNFEYHP ESEVDQQRVE MGILAKKAEL IKEEPLYNVA IAGEVEPDGV HLQVTVVVDS
QLFSQEGATH LMEQVCNTFQ ALNASL
