Ncarz_00050 : CDS information

Location

Organism	Streptomyces carzinostaticus
Strain	ATCC 15944
Entry name	Neocarzinostatin
Contig	AY117439
Start / Stop / Direction	14,674 / 15,672 / + [in whole cluster] 14,674 / 15,672 / + [in contig]
Location	14674..15672 [in whole cluster] 14674..15672 [in contig]
Type	CDS
Length	999 bp (332 aa)

Annotation

Category	3.2 modification methylation
Product	2,7-hydroxy-5-methyl-1-naphthoic acid C-7 O-methyltransferase
Product (GenBank)	O-methyltransferase
Gene	ncsB1
Gene (GenBank)
EC number	2.1.1.-
Keyword	naphthoic acid
Note
Note (GenBank)
Reference	ACC Q84HC8 PmId [15797213] The neocarzinostatin biosynthetic gene cluster from Streptomyces carzinostaticus ATCC 15944 involving two iterative type I polyketide synthases. (Chem Biol. , 2005) [18387946] Regiospecific O-methylation of naphthoic acids catalyzed by NcsB1, an O-methyltransferase involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin. (J Biol Chem. , 2008) [19702337] Molecular basis of substrate promiscuity for the SAM-dependent O-methyltransferase NcsB1, involved in the biosynthesis of the enediyne antitumor antibiotic neocarzinostatin. (Biochemistry. , 2009) [20735485] In vivo characterization of NcsB3 to establish the complete biosynthesis of the naphthoic acid moiety of the neocarzinostatin chromophore. (FEMS Microbiol Lett. , 2010) comment [PMID:15797213] Neocarzinostatin生合成遺伝子クラスターの報告。 ncsB1: O-methyl transferase C-7のヒドロキシル基をメチル化すると推測。このORFについて機能解析されていない。 [PMID:18387946] NcsB1の酵素学的解析論文。 in vitroで、O-methylation activity、反応速度論的解析、基質特異性など酵素学的解析をしている。その結果、NcsB1は2,7-dihydroxy-5-methyl-1-naphthoic acidを2-hydroxy-7-methoxy-5-methyl-1-naphthoic acidにするSAM-dependent O-methyltransferaseだとしている。しかしこの酵素は、様々なortho-hydoxynaphthoic acidsのhydroxy moietyを位置特異的にアルキル化することも可能であると示した。 [PMID:19702337] NcsB1の結晶構造解析論文。結晶構造解析により酵素活性サイトの詳細な構造を解明し、基質の結合に酵素の再配向が関与する事がわかった。また、基質認識に重要な残基を予測し、mutation株作製し生化学的な検討もしているようだ。 [PMID:20735485] ncsB3の機能解析論文。 ncsB,B1,B3をS.lividans TK24で発現。これらの産物をHPLCにて解析。種々のortho-hydroxy naphthoic acidsのhydroxy moietyを位置特異的にアルキル化できることを示した。

Sequence

>2,7-hydroxy-5-methyl-1-naphthoic acid C-7 O-methyltransferase [O-methyltransferase]
MGKRAAHIGLRALADLATPMAVRVAATLRVADHIAAGHRTAAEIASAAGAHADSLDRLLR
HLVAVGLFTRDGQGVYGLTEFGEQLRDDHAAGKRKWLDMNSAVGRGDLGFVELAHSIRTG
QPAYPVRYGTSFWEDLGSDPVLSASFDTLMSHHLELDYTGIAAKYDWAALGHVVDVGGGS
GGLLSALLTAHEDLSGTVLDLQGPASAAHRRFLDTGLSGRAQVVVGSFFDPLPAGAGGYV
LSAVLHDWDDLSAVAILRRCAEAAGSGGVVLVIEAVAGDEHAGTGMDLRMLTYFGGKERS
LAELGELAAQAGLAVRAAHPISYVSIVEMTAL

>2,7-hydroxy-5-methyl-1-naphthoic acid C-7 O-methyltransferase [O-methyltransferase]
GTGGGAAAAAGGGCTGCACACATCGGATTGAGGGCGCTGGCCGATCTGGCGACTCCCATG
GCGGTCCGCGTGGCGGCCACGCTGCGGGTCGCCGATCACATAGCGGCCGGACACCGCACC
GCCGCGGAGATCGCCTCCGCGGCCGGAGCACATGCGGATTCCCTGGACAGGTTGCTGCGC
CACCTGGTCGCCGTCGGTCTGTTCACCAGGGACGGCCAGGGCGTCTACGGTCTGACAGAG
TTTGGGGAGCAGCTGCGCGACGACCACGCTGCCGGAAAACGCAAGTGGCTGGACATGAAC
AGTGCCGTGGGGCGGGGCGACCTCGGTTTCGTCGAGTTGGCCCACAGCATTCGCACGGGG
CAGCCCGCGTACCCGGTGCGCTACGGAACGTCCTTCTGGGAGGACCTTGGCTCCGACCCC
GTTCTCTCGGCGTCCTTCGACACCCTGATGAGCCATCACCTGGAGCTGGACTACACCGGC
ATCGCAGCCAAGTACGACTGGGCTGCCCTCGGGCATGTCGTCGACGTGGGCGGCGGCAGC
GGCGGATTGCTCAGCGCACTGCTGACAGCCCACGAGGACCTGTCGGGAACCGTCCTCGAC
CTGCAAGGGCCGGCATCGGCCGCGCACCGCAGGTTTCTGGACACCGGGTTGTCCGGACGG
GCACAGGTGGTGGTGGGCAGCTTCTTCGACCCATTACCGGCCGGCGCGGGCGGCTACGTG
CTGTCTGCCGTCCTGCACGACTGGGACGACCTGTCCGCCGTGGCGATCCTGAGGCGGTGC
GCGGAGGCGGCCGGCAGCGGAGGCGTCGTGCTCGTCATCGAAGCGGTGGCCGGCGACGAG
CACGCCGGCACCGGCATGGATCTGCGCATGCTCACCTACTTCGGAGGCAAGGAACGCAGC
CTTGCCGAACTAGGCGAACTCGCCGCACAGGCCGGCCTCGCCGTACGCGCGGCCCACCCG
ATCAGCTACGTCTCGATAGTCGAAATGACCGCCCTCTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR001077 O-methyltransferase, family 2 (Domain) PF00891 Methyltransf_2 IPR011991 Winged helix-turn-helix transcription repressor DNA-binding (Domain) G3DSA:1.10.10.10 Wing_hlx_DNA_bd IPR016461 O-methyltransferase, caffeic acid-type (Family) PIRSF005739 O-mtase
SignalP	No significant hit
TMHMM	No significant hit

Ncarz_00040 Ncarz_00060