Balhi_00230 Balhi_00230   Balhi_00250 Balhi_00250

Balhi_00240 : CDS information

close this sectionLocation

Organism
StrainDSM 5908
Entry nameBalhimycin
Contig
Start / Stop / Direction51,289 / 52,479 / + [in whole cluster]
51,289 / 52,479 / + [in contig]
Location51289..52479 [in whole cluster]
51289..52479 [in contig]
TypeCDS
Length1,191 bp (396 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category2.1 modification addition of extender units
Productcytochrome P450
Product (GenBank)putative P450 monooxygenase
Gene
Gene (GenBank)oxyD
EC number
Keyword
  • Bht
Note
Note (GenBank)
Reference
ACC
PmId
[15342578] Biosynthesis of chloro-beta-hydroxytyrosine, a nonproteinogenic amino acid of the peptidic backbone of glycopeptide antibiotics. (J Bacteriol. , 2004)
[20519494] Structural characterization of OxyD, a cytochrome P450 involved in beta-hydroxytyrosine formation in vancomycin biosynthesis. (J Biol Chem. , 2010)
comment
[PMID: 15342578](2004)
oxyDはbeta-hydroxytyrosine(beta-HT)形成に関与するbhp, bpsDとオペロンを形成することから、この経路への関与を調査。

oxyDのin-frame deletion mutant株OP090は活性のある化合物を産生できなかったが、oxyD geneでの相補によりbalhimycin産生を回復する。

・oxyD deletion mutant OP090
・bhp deletion mutant OP696
・bpsD disruption mutant BpsD-cat
・null mutant JR1 (blocked in HPG and DPG synthesis)

を用いてcross-feeding実験を実施。OP090はJR1との組み合わせでのみbalhimycin産生を回復。よってOP090は OP696, BpsD-catと同じ経路、すなわちbeta-HT合成でブロックされることが実証された。

beta-HTを含んだ培地でOP090を培養するとbalhimycin産生を回復したこともふまえ、OP090がbeta-HT合成でブロックされることが明白に実証された。

---
[PMID: 20519494](2010)
発現・精製して得たOxyDとBpsD_PCP domainを滴定し、P450吸収スペクトルのシフトで基質結合を確認している。OxyDはBpsD_PCPに結合したアミノ酸を、基質として結合する。

OxyDの結晶構造を測定。この結果とアミノ酸配列から、OxyDのようなアミノ酸が結合したPCPsを酸化するP450sの共通motifを同定している。

close this sectionSequence

selected fasta
>cytochrome P450 [putative P450 monooxygenase]
MQTTNAVDLGNPDLYTTLERHARWRELAAEDAMVWSDPGSSPSGFWSVFSHRACAAVLAP
SAPLTSEYGMMIGFDRDHPDNSGGRMMVVSEHEQHRKLRKLVGPLLSRAAARKLAERVRI
EVGDVLGRVLDGEVCDAATAIGPRIPAAVVCEILGVPAEDEDMLIDLTNHAFGGEDELFD
GMTPRQAHTEILVYFDELITARRKEPGDDLVSTLVTDDDLTIDDVLLNCDNVLIGGNETT
RHAITGAVHALATVPGLLTALRDGSADVDTVVEEVLRWTSPAMHVLRVTTADVTINGRDL
PSGTPVVAWLPAANRDPAEFDDPDTFLPGRKPNRHITFGHGMHHCLGSALARIELSVVLR
VLAERVSRVDLEREPAWLRAIVVQGYRELPVRFTGR
selected fasta
>cytochrome P450 [putative P450 monooxygenase]
ATGCAGACGACGAACGCCGTCGACCTCGGCAACCCCGACCTGTACACGACCCTGGAACGG
CACGCCCGCTGGCGCGAGCTCGCGGCGGAAGACGCGATGGTGTGGAGTGACCCGGGCAGT
TCCCCCTCCGGCTTCTGGTCGGTGTTCTCGCACCGGGCGTGCGCCGCGGTCCTCGCGCCG
TCGGCGCCGCTCACCTCCGAATACGGGATGATGATCGGGTTCGACCGCGACCACCCGGAC
AACTCCGGCGGCCGGATGATGGTGGTCTCCGAACACGAGCAGCACCGCAAGCTGCGCAAG
CTCGTCGGGCCGCTGCTGTCCCGGGCGGCCGCGCGCAAGCTGGCCGAGCGGGTGCGGATC
GAGGTCGGCGACGTGCTCGGCCGGGTCCTCGACGGCGAGGTCTGCGACGCGGCCACGGCG
ATCGGCCCCCGCATCCCCGCCGCGGTCGTGTGCGAGATCCTCGGCGTGCCCGCCGAGGAC
GAAGACATGCTCATCGACCTGACCAACCACGCCTTCGGCGGCGAGGACGAGCTGTTCGAC
GGGATGACCCCGCGGCAGGCGCACACCGAGATCCTCGTCTACTTCGACGAACTGATCACC
GCGCGCCGCAAGGAACCCGGCGACGACCTCGTCAGCACGCTGGTGACCGACGACGACCTC
ACGATCGACGACGTGCTGCTCAACTGCGACAACGTGCTCATCGGCGGCAACGAGACCACG
CGGCACGCGATCACCGGCGCGGTGCACGCGCTGGCGACGGTGCCCGGCCTGCTGACGGCG
CTGCGGGACGGGAGCGCGGACGTCGACACCGTCGTGGAAGAGGTGCTGCGCTGGACCTCG
CCCGCGATGCACGTGCTCCGGGTGACGACCGCCGACGTCACGATCAACGGCCGCGACCTG
CCGTCCGGCACCCCGGTGGTGGCGTGGCTGCCCGCGGCGAACCGGGACCCCGCCGAGTTC
GACGACCCGGACACCTTCCTGCCCGGGCGGAAACCCAACCGGCACATCACCTTCGGCCAC
GGCATGCACCACTGCCTCGGGTCCGCGCTCGCGCGGATCGAGCTGTCGGTCGTGCTGCGG
GTGCTGGCCGAGCGGGTGTCCCGGGTGGACCTGGAACGGGAGCCGGCCTGGTTGCGGGCG
ATCGTCGTGCAGGGGTACCGGGAACTCCCGGTGCGGTTCACCGGGCGCTGA

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR001128 Cytochrome P450 (Family)
[235-252] 4.10000049208425e-06 PR00385 [270-281] 4.10000049208425e-06 PR00385 [336-345] 4.10000049208425e-06 PR00385 [345-356] 4.10000049208425e-06 PR00385
PR00385   P450
[20-396] 3.99999999999998e-92 G3DSA:1.10.630.10
G3DSA:1.10.630.10   Cyt_P450
[9-396] 3.19999899046355e-83 SSF48264
SSF48264   Cytochrome_P450
[269-360] 1.5e-19 PF00067
PF00067   p450
IPR002397 Cytochrome P450, B-class (Family)
[90-101] 1.60000240695997e-45 PR00359 [136-152] 1.60000240695997e-45 PR00359 [153-168] 1.60000240695997e-45 PR00359 [193-215] 1.60000240695997e-45 PR00359 [270-281] 1.60000240695997e-45 PR00359 [287-314] 1.60000240695997e-45 PR00359 [315-330] 1.60000240695997e-45 PR00359 [336-345] 1.60000240695997e-45 PR00359 [345-356] 1.60000240695997e-45 PR00359
PR00359   BP450
IPR017972 Cytochrome P450, conserved site (Conserved_site)
[338-347] PS00086
PS00086   CYTOCHROME_P450
SignalP No significant hit
TMHMM No significant hit
Balhi_00230 Balhi_00230   Balhi_00250 Balhi_00250
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