Dauno_00210 : CDS information

Location

Organism	Streptomyces sp.
Strain	C5
Entry name	Daunorubicin
Contig	U50973
Start / Stop / Direction	1,699 / 431 / - [in whole cluster] 1,498 / 2,766 / + [in contig]
Location	complement(431..1699) [in whole cluster] 1498..2766 [in contig]
Type	CDS
Length	1,269 bp (422 aa)

Annotation

Category	3.3 modification reduction
Product	cytochrome P450
Product (GenBank)	daunomycin C-14 hydroxylase
Gene
Gene (GenBank)	doxA
EC number
Keyword	C-13 hydroxylate C-13 OH group oxidation C-14 hydroxylate
Note
Note (GenBank)
Reference	ACC Q59971 PmId [8655530] Isolation and characterization of a gene from Streptomyces sp. strain C5 that confers the ability to convert daunomycin to doxorubicin on Streptomyces lividans TK24. (J Bacteriol. , 1996) [9098063] In vivo and in vitro bioconversion of epsilon-rhodomycinone glycoside to doxorubicin: functions of DauP, DauK, and DoxA. (J Bacteriol. , 1997) [9864343] Purification, properties, and characterization of recombinant Streptomyces sp. strain C5 DoxA, a cytochrome P-450 catalyzing multiple steps in doxorubicin biosynthesis. (J Bacteriol. , 1999) comment [PMID: 8655530](1996) Streptomyces sp. strain C5のdoxA同定論文。 S.lividansにおけるdoxA発現はdaunorubicin (DNR)のC-14をhydroxylateして、doxorubicin (DXR)へのin vivo生物変換をもたらす。 DoxAはDNRと13-dihydro-DNRにのみ特異的で、doxorubicin (DXR)へ変換された。 daunomycinone(DNR aglycone)、carminomycin, 13-dihydrocarminomycin, idarubicin, and aklavin は、DoxAの明らかな基質ではなかった。 -- [PMID: 9098063](1997) DoxAが 13-deoxycarminomycin → 13-dihydrocarminomycin 13-deoxy-DNR → 13-dihydro-DNR の13-hydroxylation、そしてこれに続く 13-dihydrocarminomycin → carminomycin 13-dihydro-DNR → DNR のoxidization(dehydrogenation)を触媒することも確認している。 (DauKによる4-O-methylationでcarminomycin → daunorubicin) S.lividans TK24にrhodomycin D → DXRへ変換する能力を与えるには、doxA, dauK, and dauPで必要十分であった。 -- [PMID: 9864343](1999) Recombinant DoxAの酵素活性測定。・13-deoxydaunorubicinの13-hydroxylation ・13-dihydrocarminomycinと13-dihydrodaunorubicinの13-oxidation ・daunorubicinの14-hydroxylation をDoxAは触媒した。 kcat/Km値から、これらのDNR生合成後期ステップが4-O-methyl化を受けた基質で反応が進みそうなこと、DNR→DXRへの変換は好まれた反応ではないことが示された。 DXRはDoxA活性を完全に抑制。この抑制は13-dihydro-DNRの酸化に関して競合的。よって、DNRとDXRをもっと生成し続けるには、DXRの排出が重要であると推測される。 13-deoxycarminomycin ↓[DauK] 13-deoxy-DNR ↓[DoxA] 13-dihydro-DNR ↓[DoxA] DNR ↓[DoxA] DXR
Related Reference	ACC Q9ZAU3 NITE Adria_00010 PmId [9864344] Doxorubicin overproduction in Streptomyces peucetius: cloning and characterization of the dnrU ketoreductase and dnrV genes and the doxA cytochrome P-450 hydroxylase gene. (J Bacteriol. , 1999) comment BLAST id94% Streptomyces peucetius_doxA Daunorubicin C-14 hydroxylase -- doxAについては、不活化mutantやpromoter下で相補して産物を測定、mutantでのDoxA protein発現をWBで確認。その結果からDoxAは 13-deoxy-DNR → 13-dihydro-DNR daunorubicin (DNR) → doxorubicin (DXR) への変換をするが、それにはdnrVが一緒に発現されることが必要。 13-dihydro-DNR → DNRへの変換はこの論文では確かめていない。
Related Reference	ACC Q93MI2 PmId [26334717] ( , ) comment BLAST id90% Streptomyces peucetius subsp. caesius(ATCC 27952/NBRC 14660)_doxA Cytochrome P-450 monooxygenase DoxA (EC:1.14.13.181)

Sequence

>cytochrome P450 [daunomycin C-14 hydroxylase]
MSGEAPRVAVDPFSCPMMTMQRKPEVHDAFREAGPVVEVNAPAGGPAWVITDDALAREVL
ADPRFVKDPDLAPTAWRGVDDGLDIPVPELRPFTLIAVDGEDHRRLRRIHAPAFNPRRLA
ERTDRIAAIADRLLTELADSSDRSGEPAELIGGFAYHFPLLVICELLGVPVTDPAMAREA
VGVLKALGLGGPQSAGGDGTDPAGDVPDTSALESLLLEAVHAARRKDTRTMTRVLYERAQ
AEFGSVSDDQLVYMITGLIFAGHDTTGSFLGFLLAEVLAGRLAADADGDAISRFVEEALR
HHPPVPYTLWRFAATEVVIRGVRLPRGAPVLVDIEGTNTDGRHHDAPHAFHPDRPSRRRL
TFGDGPHYCIGEQLAQLESRTMIGVLRSRFPQARLAVPYEELRWCRKGAQTARLTDLPVW
LR

>cytochrome P450 [daunomycin C-14 hydroxylase]
ATGAGCGGCGAGGCGCCGCGGGTGGCCGTCGACCCGTTCTCGTGTCCCATGATGACCATG
CAGCGCAAACCCGAGGTGCACGACGCATTCCGAGAGGCGGGCCCCGTCGTCGAGGTGAAC
GCCCCCGCGGGCGGACCCGCCTGGGTCATCACCGATGACGCCCTCGCCCGCGAGGTGCTG
GCCGATCCCCGGTTCGTGAAGGACCCCGATCTCGCGCCCACCGCCTGGCGGGGGGTGGAC
GACGGTCTCGACATCCCCGTTCCGGAGCTGCGTCCGTTCACGCTCATCGCCGTGGACGGT
GAGGACCACCGGCGTCTGCGCCGCATCCACGCACCGGCGTTCAACCCGCGCCGGCTGGCC
GAGCGGACGGATCGCATCGCCGCCATCGCCGACCGGCTGCTCACCGAACTCGCCGACTCC
TCCGACCGGTCGGGCGAACCGGCCGAGCTGATCGGCGGCTTCGCGTACCACTTCCCGCTG
TTGGTCATCTGCGAACTGCTCGGCGTGCCGGTCACCGATCCGGCAATGGCCCGCGAGGCC
GTCGGCGTGCTCAAGGCACTCGGCCTCGGCGGCCCGCAGAGCGCCGGCGGTGACGGCACG
GACCCTGCCGGGGACGTGCCGGACACGTCGGCGCTGGAGAGCCTTCTCCTCGAAGCCGTG
CACGCGGCCCGGCGGAAAGACACCCGGACCATGACCCGCGTGCTCTATGAACGCGCACAG
GCAGAGTTCGGCTCGGTCTCCGACGACCAGCTCGTCTACATGATCACCGGACTCATCTTC
GCCGGCCACGACACCACCGGCTCGTTCCTGGGCTTCCTGCTTGCGGAGGTCCTGGCGGGC
CGTCTCGCGGCGGACGCCGACGGGGACGCCATCTCCCGGTTCGTGGAGGAGGCGCTGCGC
CACCACCCGCCGGTGCCCTACACGTTGTGGAGGTTCGCTGCCACGGAGGTGGTCATCCGC
GGTGTCCGGCTGCCCCGCGGAGCGCCGGTACTGGTGGACATCGAGGGCACCAACACCGAC
GGCCGCCATCACGACGCCCCGCACGCTTTCCACCCGGACCGCCCTTCGAGGCGGCGGCTC
ACCTTCGGCGACGGGCCGCACTACTGCATCGGGGAGCAGCTCGCCCAGCTGGAATCGCGC
ACGATGATCGGCGTACTGCGCAGCAGGTTCCCCCAAGCCCGACTGGCCGTGCCGTACGAG
GAGTTGCGGTGGTGCAGGAAGGGGGCCCAGACAGCGCGGCTCACTGACCTGCCCGTCTGG
CTGCGTTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR001128 Cytochrome P450 (Family) PR00385 P450 SSF48264 Cytochrome_P450 G3DSA:1.10.630.10 Cyt_P450 PF00067 p450 IPR002397 Cytochrome P450, B-class (Family) PR00359 BP450 IPR017972 Cytochrome P450, conserved site (Conserved_site) PS00086 CYTOCHROME_P450
SignalP	No significant hit
TMHMM	No significant hit

Dauno_00220 Dauno_00200