Enter_00180 : CDS information

close this sectionLocation

Organism
Strain
Entry nameEnterocin
Contig
Start / Stop / Direction19,589 / 18,384 / - [in whole cluster]
19,589 / 18,384 / - [in contig]
Locationcomplement(18384..19589) [in whole cluster]
complement(18384..19589) [in contig]
TypeCDS
Length1,206 bp (401 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category3.3 modification reduction
Productcytochrome P450
Product (GenBank)putative p450 monooxygenase EncR
Gene
Gene (GenBank)encR
EC number
Keyword
  • C-5 hydroxylate
Note
Note (GenBank)
Reference
ACC
PmId
[11137817] Cloning, sequencing and analysis of the enterocin biosynthesis gene cluster from the marine isolate 'Streptomyces maritimus': evidence for the derailment of an aromatic polyketide synthase. (Chem Biol. , 2000)
[17704772] Enzymatic total synthesis of enterocin polyketides. (Nat Chem Biol. , 2007)
comment
[PMID: 11137817](2000)
enterocin と wailupemycinsの生合成をコードするgene clusterの同定。

EncR: Cytochrome P-450 hydroxylase

monooxygenase inhibitorであるancymidol によって、S.maritimusでのenterocin産生は減り、5-deoxyenterocinが蓄積する。

EncRをクローニング、maltose-binding-protein(MBP) affinity tagを付けて精製。反応産物のLC-MS解析から、MBP-EncRは 5-deoxyenterocin → enterocin への変換をすることを実証。

またMBP-EncRはancymidol によって抑制されたので、in vivoでのancymidol による結果が、CYP450 EncR抑制のせいであることも確認された。

---
[PMID: 17704772](2007)
enc genesを異種性に発現して得たタンパクを用いて、enterocinをin vitroに合成している。

EncA/B, EncC, EncD, EncN, EncM, EncK, EncR, SgFabD + benzoic acid, ATP/Mg2+, NADPH, SAM, ferredoxin, ferredoxin-NADP+ reductase, catalaseでenterocin, wailupemycin F and Gの産生あり。

他のCYP450でも報告があるようにEncRはSAMで抑制されることがさらなる解析から明らかになったので、5-deoxyenterocinを含むnonpolar reaction productsを酸性条件下で抽出し、これがEncR-catalyzed reactionを受けてenterocinが得られることを確認した。

close this sectionSequence

selected fasta
>cytochrome P450 [putative p450 monooxygenase EncR]
MTTHTQQLRDFPFAPPAELHMEPAFAQLREEEPISRVRLPYGGEAWLVTRYQDIKTVLGD
PRFSRAATQHAQAPRIQPDPAGEGVLMSLDPPDHTRLRKTVAGVFTKRRVEDLRPATQRI
AEELLEAMEASGAPADLVASYALPLPVTVICDLLGVPGDDREQLRGWSDALLSTTACTPA
ESAAAAQAMADHFAALVSQRRRQPTDDLLGALVQTWDREEGLLRDEELVLLTRDLLIAGH
ETTASQIANCTYLLLQRPHDMDRLRTDPSAMASAVEELLRFIPLGSGSFRARVATEPVEL
CGVRIQPGDTVFAPTVAANWDPDVFAEPGRLDIDRSPNPHVAFGHGVHHCLGAQLARLEL
QVALGVLLRRLPRLRLAVDEAEIVWKTGMQVRGPKTLPVKW
selected fasta
>cytochrome P450 [putative p450 monooxygenase EncR]
GTGACCACCCATACCCAGCAGTTGCGGGACTTTCCCTTCGCGCCCCCCGCCGAGCTGCAC
ATGGAACCGGCCTTCGCGCAATTGCGCGAAGAGGAGCCCATCAGTCGCGTCCGCCTCCCG
TACGGGGGTGAGGCATGGCTGGTCACGCGTTATCAGGACATCAAGACCGTGCTGGGCGAC
CCGCGGTTCAGTCGTGCCGCCACGCAGCACGCCCAGGCGCCGCGGATCCAGCCTGACCCG
GCGGGTGAAGGCGTGCTGATGTCGCTGGATCCCCCGGACCACACGCGGCTGCGTAAAACG
GTCGCGGGTGTCTTCACCAAGCGCAGGGTGGAAGATCTGCGGCCCGCCACGCAGAGGATC
GCCGAGGAGTTGCTGGAGGCGATGGAAGCCTCGGGTGCACCGGCCGATCTGGTCGCATCC
TATGCACTGCCGCTGCCCGTCACGGTGATCTGCGACCTGCTGGGTGTACCCGGGGACGAC
CGTGAACAGTTGCGGGGCTGGTCCGATGCGCTGCTGTCCACCACGGCCTGCACTCCAGCT
GAATCGGCGGCGGCCGCGCAGGCCATGGCCGACCACTTCGCCGCCCTGGTGAGCCAGCGG
CGCCGCCAGCCGACCGATGACCTGCTCGGGGCTCTGGTGCAGACGTGGGACCGGGAGGAG
GGGCTGCTTCGCGACGAGGAGCTGGTCCTTCTCACCCGTGATCTGCTCATCGCCGGCCAT
GAGACCACGGCGAGCCAGATCGCCAACTGCACCTACCTGCTGCTCCAGCGCCCGCATGAC
ATGGACCGCTTGCGCACGGATCCCTCGGCGATGGCGTCCGCAGTGGAGGAGCTGCTGCGG
TTCATCCCGCTGGGCTCGGGCAGTTTCCGGGCCAGGGTCGCCACCGAGCCGGTGGAACTG
TGCGGGGTGAGGATCCAGCCCGGTGACACCGTGTTCGCCCCCACTGTGGCGGCCAACTGG
GATCCGGACGTCTTTGCTGAGCCGGGCCGCCTCGATATCGACAGGTCCCCCAACCCGCAC
GTCGCCTTCGGGCACGGTGTGCACCACTGCCTGGGTGCGCAGTTGGCCAGGCTGGAGCTC
CAGGTGGCGCTGGGCGTCCTGCTGCGCCGTCTGCCGCGGCTGCGACTGGCTGTCGACGAA
GCGGAGATCGTGTGGAAGACGGGCATGCAGGTACGTGGGCCGAAGACCCTGCCGGTGAAG
TGGTGA

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09
show BLAST table
InterPro
Database:interpro:38.0
IPR001128 Cytochrome P450 (Family)
 [7-401]  6.69996745106047e-103 SSF48264
SSF48264   Cytochrome_P450
 [238-255]  1.6999992228405e-08 PR00385 [273-284]  1.6999992228405e-08 PR00385 [341-350]  1.6999992228405e-08 PR00385 [350-361]  1.6999992228405e-08 PR00385
PR00385   P450
 [187-266]  1.9e-07 PF00067 [273-370]  2e-15 PF00067
PF00067   p450
 [12-401]  1.20000000000001e-114 G3DSA:1.10.630.10
G3DSA:1.10.630.10   Cyt_P450
IPR002397 Cytochrome P450, B-class (Family)
 [89-100]  1.49999648196323e-56 PR00359 [136-152]  1.49999648196323e-56 PR00359 [153-168]  1.49999648196323e-56 PR00359 [191-213]  1.49999648196323e-56 PR00359 [273-284]  1.49999648196323e-56 PR00359 [292-319]  1.49999648196323e-56 PR00359 [320-335]  1.49999648196323e-56 PR00359 [341-350]  1.49999648196323e-56 PR00359 [350-361]  1.49999648196323e-56 PR00359
PR00359   BP450
IPR017972 Cytochrome P450, conserved site (Conserved_site)
 [343-352]  PS00086
PS00086   CYTOCHROME_P450
SignalP
 [1-26]  0.141 Signal
Bacteria, Gram-positive   
 [1-26]  0.1 Signal
Bacteria, Gram-negative   
TMHMM No significant hit
Page top