Ncarz_00080 : CDS information

Location

Organism	Streptomyces carzinostaticus
Strain	ATCC 15944
Entry name	Neocarzinostatin
Contig	AY117439
Start / Stop / Direction	22,479 / 24,155 / + [in whole cluster] 22,479 / 24,155 / + [in contig]
Location	22479..24155 [in whole cluster] 22479..24155 [in contig]
Type	CDS
Length	1,677 bp (558 aa)

Annotation

Category	3.4 other modification
Product	2-hydroxy-7-methoxy-5-methyl-1-naphthoic acid--CoA ligase 2-hydroxy-7-methoxy-5-methyl-1-naphthoyl-CoA synthetase
Product (GenBank)	adenylate ligase
Gene	ncsB2
Gene (GenBank)
EC number	6.2.1.-
Keyword	naphthoic acid
Note
Note (GenBank)
Reference	ACC Q84HC5 PmId [15797213] The neocarzinostatin biosynthetic gene cluster from Streptomyces carzinostaticus ATCC 15944 involving two iterative type I polyketide synthases. (Chem Biol. , 2005) [17539640] Characterization of NcsB2 as a promiscuous naphthoic acid/coenzyme A ligase integral to the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin. (J Am Chem Soc. , 2007) comment [PMID:15797213] Neocarzinostatin生合成遺伝子クラスターの報告。 ncsB2: CoA-ligase naphthoyl-S-NcsBからenediyne coreへnaphthoyl moietyの付着を触媒する候補であるとしている。このORFについて機能解析されていない。 [PMID:17539640] NcsB2の機能解析の報告本論文にて、naphthoic acid moietyの生合成経路がreviseされている。また、orf27をNcsB4と改名している。 in vivo, in vitroでNcsB2の機能解析をしている。 CoA ligase活性をATP-[32P]pyrophosphate exchange assay、および、Coenzyme A arylation assayを確認している。また、酵素反応速度論的解析から、ncsB2は最適基質を2-hydroxy-7-methoxy-5-methyl-1-naphthoic acidであると同定している。Kcat/Km値は低くなるが2-hydroxy-5-methyl-1-naphthoic acidや2,7-dihydroxy-5-methyl-1-naphthoic acidも活性化した。

Sequence

>2-hydroxy-7-methoxy-5-methyl-1-naphthoic acid--CoA ligase [adenylate ligase]
MHETAAAPAPAGFVPWPDDVAARYTAAGHWEGRSLGTHLAEAARKVPEAVCLVDGPVRMS
YSELMARADGAAVRMRGLGIRPADRVVVQLPNCWEHVVVTMACLRLGALPIWALPQYRHR
ELSGVVTHARASALVVPDVYREFDHQALAHEVAEAQPTVRHVLVAGSDVRPDSVDLRALC
EPLDADEAARVAAELDRSAPRGEEVAMLKLSGGTTGLPKLVARTHNDLSYMIKRAAQVCG
FGRDTVYLAVLPLGHGFPNTGPGVLGTLLAGGRVVISGSPAPEAAFALMERERVTATSVV
PAIVMRWLQYRDERPGADLGSLELMQVGASRLEPEVARQVGPKLGCRLQQVFGMAEGLLC
LTRLDDPDDVVHYTQGRPISPDDEIRVVDPEGRTVGVGEPGALLTRGPYTPRGYYDSPSA
NARAFTPDGWYRTGDLVRRTPDGNLIVVGREKDLINRGGEKINAEEVEGFAVQVDGVLQA
AAVGLPDSELGERICLFVVLADGTRVELADVRKVMENAETASFKLPERLITLPSLPTTPM
GKIDKKALRAAAGRMSET

>2-hydroxy-7-methoxy-5-methyl-1-naphthoic acid--CoA ligase [adenylate ligase]
ATGCACGAGACAGCAGCAGCACCGGCCCCCGCAGGATTCGTCCCCTGGCCCGACGATGTG
GCAGCCCGCTATACGGCCGCCGGCCACTGGGAGGGACGCTCGCTGGGGACGCACCTGGCG
GAGGCCGCACGGAAGGTCCCAGAGGCCGTATGCCTCGTCGACGGTCCGGTCCGTATGAGC
TACAGCGAGTTGATGGCACGCGCGGACGGGGCGGCCGTCCGTATGAGAGGGCTCGGCATT
CGCCCGGCCGACCGGGTGGTGGTGCAGCTACCCAACTGCTGGGAGCACGTGGTCGTGACC
ATGGCGTGCCTGCGGCTCGGCGCTCTGCCGATCTGGGCGCTGCCGCAGTACCGGCACCGC
GAACTGTCGGGCGTGGTGACTCACGCCCGGGCCAGTGCCCTCGTCGTGCCCGACGTGTAC
CGGGAGTTCGACCACCAGGCGCTGGCGCACGAAGTGGCGGAGGCGCAGCCGACTGTCCGG
CACGTCCTCGTGGCGGGGTCCGACGTACGGCCGGACAGCGTCGATCTGCGCGCGCTGTGC
GAGCCTCTGGACGCGGACGAGGCCGCGCGGGTCGCGGCCGAGCTCGACCGGTCCGCGCCG
CGTGGTGAGGAGGTCGCGATGCTCAAGCTGTCCGGCGGGACGACCGGACTGCCCAAGCTC
GTGGCACGCACGCACAACGACCTCAGCTACATGATCAAGCGCGCTGCCCAGGTGTGCGGC
TTCGGACGGGACACGGTGTACTTGGCCGTCCTGCCGCTCGGTCACGGCTTCCCCAACACC
GGTCCCGGAGTCCTCGGCACGCTCCTGGCCGGGGGGCGGGTGGTGATTTCCGGATCGCCG
GCTCCCGAGGCCGCTTTCGCGCTGATGGAGCGGGAACGGGTGACCGCCACGTCCGTGGTG
CCGGCCATTGTCATGCGGTGGCTCCAGTACCGGGACGAGCGGCCGGGCGCCGACCTGGGT
TCGCTGGAGCTCATGCAGGTCGGGGCCTCGCGGCTCGAGCCGGAGGTGGCCCGGCAGGTG
GGACCGAAACTGGGCTGCAGGCTGCAGCAGGTGTTCGGGATGGCGGAGGGCCTGCTGTGC
CTGACCCGCCTGGACGATCCGGACGACGTGGTCCACTACACCCAGGGACGGCCGATCAGC
CCGGACGACGAGATCCGGGTGGTGGACCCGGAGGGCAGGACAGTGGGTGTGGGCGAACCC
GGGGCGTTGCTCACGCGGGGGCCGTACACGCCGCGCGGCTACTACGATTCGCCCTCCGCC
AACGCGCGAGCGTTCACCCCCGACGGCTGGTACCGGACCGGTGACCTGGTACGCCGGACG
CCGGACGGCAACCTGATCGTGGTCGGCAGGGAGAAGGACCTCATCAACAGGGGTGGCGAG
AAGATCAACGCCGAGGAGGTGGAGGGCTTCGCGGTGCAGGTCGACGGAGTTCTGCAGGCG
GCTGCCGTCGGCCTGCCGGACAGCGAACTCGGCGAGCGGATCTGTCTGTTCGTCGTGCTG
GCCGACGGCACGCGGGTGGAGCTGGCGGACGTCCGGAAGGTGATGGAGAACGCCGAGACG
GCGTCGTTCAAGCTTCCGGAGCGTCTCATCACGCTTCCCTCCCTGCCCACCACACCGATG
GGCAAGATCGACAAGAAGGCACTGCGTGCTGCCGCCGGTCGGATGTCCGAAACCTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR000873 AMP-dependent synthetase/ligase (Domain) PF00501 AMP-binding IPR020845 AMP-binding, conserved site (Conserved_site) PS00455 AMP_BINDING
SignalP	No significant hit
TMHMM	No significant hit

Ncarz_00070 Ncarz_00090