Chlor_00270 : CDS information

Location

Organism	Amycolatopsis orientalis
Strain	A82846UV37B
Entry name	Chloroeremomycin
Contig	AJ223998
Start / Stop / Direction	54,868 / 55,485 / + [in whole cluster] 20,624 / 21,241 / + [in contig]
Location	54868..55485 [in whole cluster] 20624..21241 [in contig]
Type	CDS
Length	618 bp (205 aa)

Annotation

Category	2.3 modification addition of sugar moiety
Product	dTDP-3-amino-4-keto-2,3,6-trideoxy-3-C-methyl-D-glucose 5-epimerase
Product (GenBank)	PCZA361.16
Gene	evaD ORF26
Gene (GenBank)
EC number	5.1.3.-
Keyword	L-4-epivancosamine
Note
Note (GenBank)	similar to epimerase
Reference	ACC O52806 PmId [9545426] Sequencing and analysis of genes involved in the biosynthesis of a vancomycin group antibiotic. (Chem Biol. , 1998) [11035791] Deoxysugars in glycopeptide antibiotics: enzymatic synthesis of TDP-L-epivancosamine in chloroeremomycin biosynthesis. (Proc Natl Acad Sci U S A. , 2000) [12077451] Purification, crystallization and preliminary structural studies of dTDP-4-keto-6-deoxy-glucose-5-epimerase (EvaD) from Amycolatopsis orientalis, the fourth enzyme in the dTDP-L-epivancosamine biosynthetic pathway. (Acta Crystallogr D Biol Crystallogr. , 2002) [15159413] The position of a key tyrosine in dTDP-4-Keto-6-deoxy-D-glucose-5-epimerase (EvaD) alters the substrate profile for this RmlC-like enzyme. (J Biol Chem. , 2004) comment [PMID: 9545426](1998) chloroeremomycin生合成 gene clusterの報告。 ORF26(205aa): 3,5-Epimerase 配列解析のみ。NDP-4-epi-vancosamineの生合成への関与が割り当てられている。 --- [PMID: 11035791](2000) dTDP-4-keto-6-deoxy-D-glucoseからdTDP-L-epivancosamineを生合成する5つの酵素を、E.coliで発現・精製し、特徴づけしている。 ORF26でdTDP-3-amino-2,3,6-trideoxy-3-C-methyl-D-erythro-hexopyranos-4-uloseを反応させると、分子量は変わらないがchromatographyだと明らかに異なる化合物をもたらしたことから、ORF26はC-5 epimeraseとして機能すると予測された。C-5での重水素の取り込みで、これを確認している。すべての結果から提唱されている経路は次の通り。 dTDP-4-keto-6-deoxy-D-glucose ↓ ORF23(EvaA): C-2 deoxygenation ↓ ORF25(EvaB): C-3 amination dTDP-3-amino-2,3,6-trideoxy-D-threo-hexopyranos-4-ulose ↓ ORF14(EvaC): C-3 methylation dTDP-3-amino-2,3,6-trideoxy-3-C-methyl-D-erythro-hexopyranos-4-ulose ↓ ORF26(EvaD): C-5 epimerization ↓ ORF24(EvaE): C-4 ketoreduction dTDP-L-epivancosamine --- [PMID: 12077451] (2002) dTDP-3-amino-4-keto-2,3,6-trideoxy-3-C-methyl-glucose-5-epimerase (EvaD)の結晶構造解析。 --- [PMID: 15159413](2004) 結晶構造解析のグループの続報。EvaDの構造解析と生化学的特徴づけ。 EvaDはRmlCクラスの酵素なのにdouble epimerization活性がとても低いのは、触媒残基Tyrの位置に関係することをmutant作成などで確認している。

Sequence

>dTDP-3-amino-4-keto-2,3,6-trideoxy-3-C-methyl-D-glucose 5-epimerase [PCZA361.16]
MQARKLAVDGAIEFTPRVFADDRGLLILPYQEEAFVEAHGGPLFRVAQTIHSMSKRGVVR
GIHYTVTPPGTAKYVYCARGKAMDIVIDIRVGSPTFGQWDSVLMDQQDPRAVYLPVGVGH
AFVALEDDTVMSYMLSRSYVTQDELALSALDPALGLPIDIGVEPIVSDRDRVAITLAEAQ
RQGLLPDYTTSQEIERRLTAVPVST

>dTDP-3-amino-4-keto-2,3,6-trideoxy-3-C-methyl-D-glucose 5-epimerase [PCZA361.16]
GTGCAAGCACGCAAACTCGCCGTCGACGGCGCGATCGAGTTCACCCCTCGCGTCTTCGCC
GACGACCGGGGCCTGTTGATCCTGCCGTACCAGGAAGAGGCCTTCGTCGAGGCGCACGGC
GGCCCGCTTTTCCGTGTGGCACAGACGATCCACAGCATGTCCAAGCGCGGCGTGGTGCGG
GGCATCCACTACACCGTGACGCCACCGGGTACCGCCAAATACGTCTACTGCGCCCGCGGC
AAGGCCATGGACATCGTGATCGACATCAGAGTCGGCTCGCCCACGTTCGGGCAGTGGGAC
TCGGTGCTGATGGACCAGCAGGATCCCCGGGCGGTGTACCTTCCGGTCGGCGTCGGCCAT
GCGTTCGTGGCCCTCGAGGACGACACCGTCATGTCGTACATGCTCTCCAGAAGCTATGTG
ACGCAGGACGAACTCGCCCTCTCGGCCCTGGATCCCGCACTGGGCCTGCCCATCGACATC
GGCGTCGAACCGATCGTGTCCGATCGGGACCGGGTGGCCATCACGCTCGCCGAGGCCCAG
CGGCAGGGCCTGCTGCCGGATTACACCACCAGTCAGGAGATCGAGCGGCGGCTGACCGCG
GTCCCCGTCTCGACCTGA

Feature

BLASTP Database:UniProtKB:2011_09	show BLAST table
InterPro Database:interpro:38.0	IPR000888 dTDP-4-dehydrorhamnose 3,5-epimerase-related (Family) PD001462 dTDP_sugar_isom PF00908 dTDP_sugar_isom IPR011051 RmlC-like cupin domain (Domain) SSF51182 RmlC_like_cupin IPR014710 RmlC-like jelly roll fold (Domain) G3DSA:2.60.120.10 RmlC-like_jellyroll
SignalP	No significant hit
TMHMM	No significant hit

Chlor_00260 Chlor_00280