Gilvo_00230 Gilvo_00230   Gilvo_00250 Gilvo_00250

Gilvo_00240 : CDS information

close this sectionLocation

Organism
StrainGö 3592
Entry nameGilvocarcin
Contig
Start / Stop / Direction27,551 / 28,483 / + [in whole cluster]
27,551 / 28,483 / + [in contig]
Location27551..28483 [in whole cluster]
27551..28483 [in contig]
TypeCDS
Length933 bp (310 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category1.1 PKS
Productpolyketide synthase acyltransferase subunit
malonyl-CoA--acyl carrier protein transacylase
Product (GenBank)putative malonyl-CoA acyl carrier protein transacylase
Gene
Gene (GenBank)gilP
EC number
Keyword
  • type II PKS
  • malonyl-CoA
  • acetyl-CoA
Note
Note (GenBank)
  • GilP
Reference
ACC
PmId
[12822997] The complete gene cluster of the antitumor agent gilvocarcin V and its implication for the biosynthesis of the gilvocarcins. (J Am Chem Soc. , 2003)
[20617244] Delineating the earliest steps of gilvocarcin biosynthesis: role of GilP and GilQ in starter unit specificity. (Org Biomol Chem. , 2010)
[20486694] Enzymatic total synthesis of rabelomycin, an angucycline group antibiotic. (Org Lett. , 2010)
comment
[PMID: 12822997](2003)
gilvocarcin(gil) gene cluster同定のためにクローニングされたcos-G9B3をS.lividans TK24に導入すると、wildレベルのgilvocarcins V and M産生あり。よってgilvocarcin生合成のための完全なclusterが含まれている。

cos-G9B3をサブクローニングしてシークエンス。32.9kbpに26ORFsあり。
supporting info. Table 1
GilP(310aa): Malonyl CoA-acyl carrier protein transacylase

配列解析のみ

--
[PMID: 20617244](2010)
GilP, GilQについて検討。

>in vivo実験
GilP不活化mutantの産物蓄積はwildと変わりない。

>in vitro実験
E.coliでGilP, GilQ, RavC(ACPであるGilCがうまく得られなかったので代替), Svp(RavCをapo→holoにするPPTase)を発現、精製。
14C-acyl-CoAsとの特異的活性を測定。
GilPは、malonyl-CoA > methylmalonyl-CoA > propionyl-CoA > acetyl-CoA
GilQは、propionyl-CoA > malonyl-CoA > acetyl-CoA = methylmalonyl-CoA

>配列解析から予測されたactive siteの検討
GilP(S90A)は14C-malonyl-CoA、GilQ(S111A)は14C-propionyl-CoAをロードできない。
RavC(S39A)も14C-malonyl-CoAのロードが抑制された。

gilvocarcin minimal PKS + GilPは、malonyl-CoA and acetyl-CoAの両方が存在していないとacetate primed gilvocarcin M(GM)の中間体を産生可能という報告あり(ref28=PMID: 20486694)。
また、decarboxylationに必要な残基がgil KS-beta_GilBで置換されている。
よってGilPは直接holo-ACPへacetyl-CoAを取り込むと考えられる。

GMはminor産物であり、GilPの主な役割は伸長units malonyl-CoAの動員と移動である。

close this sectionPKS/NRPS Module

AT4..254

close this sectionSequence

selected fasta
>polyketide synthase acyltransferase subunit [putative malonyl-CoA acyl carrier protein transacylase]
MRAFLFPGQGTQKIGMGTYLRERYPHLIAPLWREADDVLGFPLTRLCEEGPGEKLRHMPV
TQPAVFLCSYAALVAAQANGAEPDVIAGHSLGEYSALAAAGVLTWQEVLQLVHRRGQLMA
EVQHKVDGKMAAVIGLAIGQVEEICEQVRSETGEVVEVANHNEPLQVVVSGQCAAIDLLV
QRVATATDVRTSVLRIGGPAHSSLMGSVAGDFVEYLRRFDFCTPKTMLISGSTAEPYASA
EEIRHQLGRQLVHRVRWVDVMAQLERLGVAQTWELGPGKVLSGFVQRSLPQVRTYRANDL
PSFLAGVTGW
selected fasta
>polyketide synthase acyltransferase subunit [putative malonyl-CoA acyl carrier protein transacylase]
GTGAGAGCGTTCCTGTTCCCCGGTCAGGGGACCCAGAAGATCGGCATGGGCACCTACCTG
CGAGAACGGTACCCCCACCTGATCGCGCCGTTGTGGCGGGAGGCGGACGACGTCCTGGGT
TTCCCCCTCACCCGCCTCTGCGAGGAAGGCCCCGGCGAGAAGCTCCGCCACATGCCGGTC
ACCCAGCCCGCCGTCTTCCTGTGCAGTTACGCCGCGCTCGTCGCCGCGCAGGCGAACGGC
GCGGAGCCGGACGTCATCGCGGGCCACAGTCTGGGCGAGTACTCGGCGCTGGCGGCGGCC
GGCGTCCTCACCTGGCAGGAGGTCCTTCAGCTCGTCCACCGCCGCGGTCAGCTCATGGCG
GAGGTGCAGCACAAGGTGGACGGGAAGATGGCGGCCGTCATCGGTCTCGCCATCGGGCAG
GTCGAGGAGATCTGCGAGCAGGTGCGGTCCGAGACCGGTGAGGTGGTCGAGGTGGCCAAC
CACAACGAGCCCCTCCAGGTCGTCGTCTCCGGCCAGTGCGCTGCCATAGACCTCCTGGTC
CAGCGCGTCGCGACGGCGACCGACGTCCGCACGTCCGTCCTGAGGATCGGTGGCCCGGCC
CACTCCAGTCTCATGGGCAGCGTCGCGGGGGACTTCGTGGAGTACCTCCGGCGCTTCGAC
TTCTGCACGCCCAAGACGATGCTGATCTCCGGGTCGACCGCCGAGCCCTACGCGAGTGCG
GAGGAGATCAGGCACCAGCTCGGCAGGCAGCTGGTGCACCGGGTGCGGTGGGTGGACGTG
ATGGCGCAGCTCGAGAGGCTGGGGGTCGCACAGACCTGGGAGCTGGGGCCGGGCAAGGTC
CTCTCGGGATTCGTACAGCGGTCGCTGCCTCAGGTGCGGACGTACCGCGCGAATGATCTG
CCGTCCTTCCTGGCCGGCGTGACGGGCTGGTGA
AT4..254
AT10..762

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR001227 Acyl transferase domain (Domain)
[3-122] 3.99999999999998e-56 G3DSA:3.40.366.10 [198-295] 3.99999999999998e-56 G3DSA:3.40.366.10
G3DSA:3.40.366.10   Ac_transferase_reg
IPR004410 Malonyl CoA-acyl carrier protein transacylase, FabD-type (Family)
[2-288] 1.29999999999998e-79 TIGR00128
TIGR00128   FabD
IPR014043 Acyl transferase (Domain)
[4-254] 7.89999999999997e-37 PF00698
PF00698   Acyl_transf_1
IPR016035 Acyl transferase/acyl hydrolase/lysophospholipase (Domain)
[1-303] 7.40000780398342e-60 SSF52151
SSF52151   Acyl_Trfase/lysoPlipase
IPR016036 Malonyl-CoA ACP transacylase, ACP-binding (Domain)
[126-197] 1.09999909120787e-13 SSF55048
SSF55048   Malonyl_transacylase_ACP-bd
IPR024925 Malonyl CoA-acyl carrier protein transacylase (Family)
[1-307] 2.1000026783403e-63 PIRSF000446
PIRSF000446   Mct
SignalP No significant hit
TMHMM No significant hit
Gilvo_00230 Gilvo_00230   Gilvo_00250 Gilvo_00250
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