Stref_00200 Stref_00200   Stref_00220 Stref_00220

Stref_00210 : CDS information

close this sectionLocation

Organism
StrainNRRL 3193 (=NBRC 13446)
Entry nameSteffimycin
Contig
Start / Stop / Direction19,995 / 21,203 / + [in whole cluster]
19,995 / 21,203 / + [in contig]
Location19995..21203 [in whole cluster]
19995..21203 [in contig]
TypeCDS
Length1,209 bp (402 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category3.3 modification reduction
Productputative cytochrome P450
Product (GenBank)cytochrome P450
Gene
Gene (GenBank)stfPI
EC number
Keyword
Note
Note (GenBank)
Reference
ACC
PmId
[16751529] Isolation, characterization, and heterologous expression of the biosynthesis gene cluster for the antitumor anthracycline steffimycin. (Appl Environ Microbiol. , 2006)
comment
steffimycin生合成gene clusterのクローニング、特徴づけ。
36ORFsが見つかり、そのうち24ORFsを含む26.5 kb領域がおそらくsteffimycin生合成に関連する。

stfPI: Monooxygenase

配列解析から、cytochrome P450 StfPI and dehydrogenase StfEはおそらくC-10 keto基形成に関与すると提唱されている。

酵素反応の実験確認はないが、各中間体が検出されていることから、
L-rhamnose付着 → L-rhamnose C-2' methoxy基形成 → C-10 keto基形成 → C-8 methoxy基形成 = steffimycin
という順番であることは確かである。
Related Reference
ACC
P48635
PmId
[8416893] Identification of a Saccharopolyspora erythraea gene required for the final hydroxylation step in erythromycin biosynthesis. (J Bacteriol. , 1993)
[7849045] ( , )
comment
Blast 29th, id40%, 1e-64
Saccharopolyspora erythraea_eryK
Erythromycin C-12 hydroxylase(EC 1.14.13.154)

---
[PMID: 8416893](1993)
Saccharopolyspora erythraea (Streptomyces erythraeus)において、P450 monooxygenase familyに属するEryKが、erythromycin生合成の最終反応の１つであるmacrolactone ringのC-12 hydroxylationに関与していることをeryK変異体を作成して調べている。

---
[PMID: 7849045](1995)
不溶性封入体としてE.coliでEryKを過剰産生し、可溶化、再折りたたみ、active holo-EryK再構築をした。動態確認で、erythromycin Bより1200-1900倍もerythromycin Dを基質として好むことが分かった。

close this sectionSequence

selected fasta
>putative cytochrome P450 [cytochrome P450]
MTRGAVSMGGPHVSDEVREMLDRYRQMRRTEPVSQDKDTGRWQVFRYGDGMTVATDANTF
SNDLSEFIPDHEELRTFSKGNFQNMDQPRHRELRGLVSQGFTPRFVAGMAPRIETVAKEL
LDALRDRDRIDLVTDLAQPLPVTVIAEVMGLPVSDRALFRKWADALLVDNGIDAIFSEQG
VAAMAPAIREMNAYLLNHVREYRTSDREGLIGDLIAAETDGMTLDDQEIVGFLALLLIAG
HLTTSTLLSNAILCLDENPDATRQLRDDPGLLPNAIEEVLRYRSPLFWLDRRARETVELG
GCEIPAGSQVSVALASVSRDEEEFENPDTFDIHRNPVRHLAFGMGAHFCLGAPLARLEVR
IALKALLDRYRDFAVASDEPIVYHDPRGFQGATHLPVDLLVA
selected fasta
>putative cytochrome P450 [cytochrome P450]
GTGACGCGAGGAGCTGTTTCGATGGGTGGACCGCACGTCTCCGACGAAGTGCGCGAGATG
CTGGACCGGTACCGGCAGATGCGCCGGACCGAGCCGGTCAGCCAGGACAAGGACACCGGA
CGGTGGCAGGTGTTCCGCTATGGCGACGGGATGACCGTCGCCACCGACGCGAACACGTTC
TCCAACGATCTCAGCGAGTTCATTCCCGACCACGAGGAACTCAGGACGTTCTCCAAGGGG
AACTTCCAGAACATGGACCAGCCGCGCCACCGGGAGTTGCGCGGCCTGGTCAGCCAGGGG
TTCACGCCCCGGTTCGTCGCGGGCATGGCCCCGCGCATCGAGACGGTGGCCAAGGAGTTG
CTGGACGCGCTGCGAGACCGTGACCGTATCGACCTGGTGACCGACCTCGCCCAGCCCCTG
CCGGTCACCGTCATCGCGGAGGTGATGGGGCTGCCGGTCTCCGACCGCGCCCTGTTCCGC
AAGTGGGCGGACGCGTTGCTGGTGGACAACGGCATCGACGCCATCTTCAGCGAGCAGGGC
GTGGCCGCGATGGCCCCGGCGATCCGCGAAATGAACGCCTATCTGCTCAATCACGTCCGC
GAGTACCGCACGTCGGACCGAGAGGGTCTCATCGGGGACCTCATCGCCGCGGAGACCGAC
GGAATGACGCTGGACGACCAGGAGATCGTGGGTTTCCTCGCCCTGCTGCTGATCGCCGGC
CATCTCACCACCAGCACGCTGCTCAGCAATGCGATCCTCTGCCTGGACGAGAACCCGGAC
GCGACACGGCAGTTGCGCGACGACCCCGGACTGCTCCCCAACGCGATCGAGGAGGTCCTG
CGCTATCGGTCTCCCCTGTTCTGGCTCGACCGCAGGGCACGCGAGACGGTCGAGCTCGGC
GGGTGCGAGATACCGGCGGGCTCACAGGTCAGCGTGGCCCTGGCCTCGGTCTCCCGGGAC
GAGGAGGAGTTCGAGAACCCGGACACCTTCGACATCCACCGCAACCCGGTCCGCCATCTG
GCCTTCGGCATGGGCGCGCACTTCTGCCTCGGCGCGCCGCTCGCCAGGCTGGAGGTGAGG
ATCGCACTCAAGGCTCTGCTGGACAGGTACCGGGACTTCGCCGTGGCGTCCGACGAGCCG
ATCGTCTACCACGACCCACGCGGATTCCAGGGAGCCACGCACCTCCCGGTCGACCTGCTC
GTCGCGTGA

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR001128 Cytochrome P450 (Family)
[239-256] 5.39999977351167e-06 PR00385 [274-285] 5.39999977351167e-06 PR00385 [340-349] 5.39999977351167e-06 PR00385 [349-360] 5.39999977351167e-06 PR00385
PR00385   P450
[191-267] 1.3e-06 PF00067 [271-371] 6.60000000000002e-24 PF00067
PF00067   p450
[18-399] 9.40003071030835e-91 SSF48264
SSF48264   Cytochrome_P450
[15-399] 2.90000000000003e-107 G3DSA:1.10.630.10
G3DSA:1.10.630.10   Cyt_P450
IPR002397 Cytochrome P450, B-class (Family)
[85-96] 1.20000117458134e-49 PR00359 [131-147] 1.20000117458134e-49 PR00359 [148-163] 1.20000117458134e-49 PR00359 [193-215] 1.20000117458134e-49 PR00359 [274-285] 1.20000117458134e-49 PR00359 [291-318] 1.20000117458134e-49 PR00359 [319-334] 1.20000117458134e-49 PR00359 [340-349] 1.20000117458134e-49 PR00359 [349-360] 1.20000117458134e-49 PR00359
PR00359   BP450
IPR017972 Cytochrome P450, conserved site (Conserved_site)
[342-351] PS00086
PS00086   CYTOCHROME_P450
SignalP No significant hit
TMHMM No significant hit
Stref_00200 Stref_00200   Stref_00220 Stref_00220
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