Spino_00150 : CDS information

Location

Organism	Saccharopolyspora spinosa
Strain	NRRL 18538
Entry name	Spinosyn
Contig	AY007564
Start / Stop / Direction	19,713 / 18,541 / - [in whole cluster] 19,713 / 18,541 / - [in contig]
Location	complement(18541..19713) [in whole cluster] complement(18541..19713) [in contig]
Type	CDS
Length	1,173 bp (390 aa)

Annotation

Category	2.3 modification addition of sugar moiety
Product	glycosyltransferase
Product (GenBank)	probable NDP-rhamnosyltransferase
Gene
Gene (GenBank)	spnG
EC number
Keyword	L-rhamnose
Note
Note (GenBank)	involved in rhamnose addition
Reference	ACC Q9ALM8 PmId [11358695] Cloning and analysis of the spinosad biosynthetic gene cluster of Saccharopolyspora spinosa. (Chem Biol. , 2001) [11386361] A cluster of genes for the biosynthesis of spinosyns, novel macrolide insect control agents produced by Saccharopolyspora spinosa. (Antonie Van Leeuwenhoek. , 2000) [19126547] Functional characterization and substrate specificity of spinosyn rhamnosyltransferase by in vitro reconstitution of spinosyn biosynthetic enzymes. (J Biol Chem. , 2009) [21544146] Enzyme-catalysed [4+2] cycloaddition is a key step in the biosynthesis of spinosyn A. (Nature. , 2011) [22283226] Structural studies of the spinosyn rhamnosyltransferase, SpnG. (Biochemistry. , 2012) comment [PMID: 11358695](2001) spinosad biosynthetic gene clusterの報告。 spnG(391aa): rhamnosyltransferase aglyconeに対してrhamnoseを付加する。 spnG mutantでの産物なし、rhamnoseの付いたpseudoaglyconeからはspinosyn合成可能。 -- [PMID: 11386361](2000) 同じ著者らの先行論文。 spnG: Rhamnose attachment --- [PMID: 19126547](2009) spnGをE.coli発現、精製、基質特異性の確認、活性測定、系統解析。 Spinosyn rhamnosyltransferase (SpnG)の基質特異性はとてもゆるい。 --- [PMID: 21544146](2011) 使用している株はNRRL 18537. SpnF, SpnL, and SpnMについての機能検証論文。 3環aglycone 8 + SpnG + dTDP-beta-L-rhamnoseで、rhamnoseの付いた3環中間体10が形成される。 3環aglycone 8 + SpnG + dTDP-beta-L-rhamnose + SpnLで、rhamnoseの付いた4環中間体が得られる。よって、SpnGは4環aglycone完成の前にrhamnoseを付加して3環中間体10を形成。その後SpnLによってC-3 and C-14間の最終環化が触媒される。 --- [PMID: 22283226](2012) 使用している株はNRRL 18537. SpnG、dTDPに結合したSpnG、aglyconeに結合したSpnGの構造測定。 SpnGはdTDP-sugars と UDP-sugarsとの識別にN202を利用する。 SpnGとspinosyn aglyconeの結合は疎水性相互作用を介して媒介される。

Sequence

>glycosyltransferase [probable NDP-rhamnosyltransferase]
MRVLVVPLPYPTHLMAMVPLCWALQASGHEVLIAAPPELQATAHGAGLTTAGIRGNDRTG
DTGGTTQLRFPNPAFGQRDTEAGRQLWEQTASNVAQSSLDQLPEYLRLAEAWRPSVLLVD
VCALIGRVLGGLLDLPVVLHRWGVDPTAGPFSDRAHELLDPVCRHHGLTGLPTPELILDP
CPPSLQASDAPQGAPVQYVPYNGSGAFPAWGAARTSARRVCICMGRMVLNATGPAPLLRA
VAAATELPGVEAVIAVPPEHRALLTDLPDNARIAESVPLNLFLRTCELVICAGGSGTAFT
ATRLGIPQLVLPQYFDQFDYARNLAAAGAGICLPDEQAQSDHEQFTDSIATVLGDTGFAS
AAIKLSDEITAMPHPAALVRTLENTAAIRA

>glycosyltransferase [probable NDP-rhamnosyltransferase]
ATGCGCGTACTCGTCGTTCCCTTGCCCTATCCGACGCATCTCATGGCAATGGTGCCGCTG
TGCTGGGCGCTGCAAGCATCCGGGCACGAGGTCCTGATCGCCGCACCACCAGAGCTGCAG
GCGACCGCGCATGGTGCAGGTCTCACCACGGCCGGGATCCGCGGGAACGACAGGACCGGC
GATACGGGTGGAACCACGCAGCTGCGCTTTCCCAATCCGGCGTTCGGTCAGCGCGACACC
GAGGCAGGCCGGCAACTGTGGGAGCAGACCGCGTCCAATGTCGCGCAAAGCTCGCTCGAT
CAGCTCCCCGAATACCTTCGACTGGCCGAGGCCTGGCGACCGTCAGTGCTGTTGGTCGAC
GTCTGCGCGCTGATCGGCCGGGTGCTCGGCGGATTGCTCGACCTGCCGGTCGTGCTGCAC
CGCTGGGGAGTCGACCCCACCGCAGGCCCCTTCAGCGATCGAGCCCACGAGTTGCTTGAC
CCGGTGTGCCGGCACCACGGACTGACCGGCCTGCCCACTCCCGAGCTCATCCTCGATCCC
TGTCCGCCGAGCCTGCAAGCAAGCGACGCGCCGCAAGGCGCACCGGTCCAGTACGTGCCG
TACAACGGAAGCGGCGCATTCCCGGCATGGGGCGCGGCGCGCACCTCAGCACGGCGGGTC
TGCATCTGCATGGGCCGCATGGTGCTGAACGCCACCGGGCCGGCTCCGCTGCTGCGCGCA
GTAGCGGCTGCCACCGAGTTGCCCGGCGTCGAGGCCGTGATCGCCGTTCCCCCCGAGCAC
CGGGCACTTCTCACCGACCTACCCGACAACGCCCGGATCGCCGAATCGGTCCCGCTCAAC
CTGTTCCTGCGTACCTGCGAGCTGGTCATCTGCGCGGGCGGCTCGGGAACGGCATTCACC
GCGACCCGACTCGGCATCCCGCAACTCGTGCTTCCCCAGTACTTCGACCAGTTCGACTAC
GCGCGCAACCTCGCCGCTGCCGGGGCGGGCATCTGCTTGCCGGATGAGCAGGCCCAGTCC
GACCACGAACAGTTCACCGACTCAATCGCAACGGTGCTCGGCGACACCGGCTTCGCTTCT
GCGGCAATCAAACTCAGCGACGAGATCACGGCCATGCCCCATCCCGCCGCGCTGGTGCGG
ACACTGGAGAACACTGCGGCCATCCGTGCCTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR004276 Glycosyl transferase, family 28 (Domain) PF03033 Glyco_transf_28 IPR010610 Domain of unknown function DUF1205 (Domain) PF06722 DUF1205
SignalP	Eukaryota Bacteria, Gram-negative Bacteria, Gram-positive
TMHMM	No significant hit

Spino_00140 Spino_00160