Cloning and Heterologous Expression of the Aranciamycin Biosynthetic Gene Cluster Revealed a New Flexible Glycosyltransferase

Abstract

Add sugar…︁ Cloning and heterologous expression of the aranciamycin biosynthetic gene cluster revealed a new flexible glycosyltransferase, AraGT, which accepts different nucleotide‐activated sugars (D‐amicetose, L‐rhodinose, L‐rhamnose and L‐axenose) The newly generated aranciamycin derivatives displayed antiproliferative activity against MaTu and MCF7 cells; this shows that the deoxysugar is important for anticancer activity.

Formation and Attachment of the Deoxysugar Moiety and Assembly of the Gene Cluster for Caprazamycin Biosynthesis

“…Cpz31, together with most glycosyltransferases involved in natural product biosynthesis, belongs to the GT-1 family of glycosyltransferases as classified by the CAZy system (6). Cpz31 exhibits significantly higher homology to, for example, the 4,6-dideoxy-4-hydroxylamino-␣-Dglucosyltransferase CalG3 (36% identity, 50% similarity) from Micromonospora echinospora (59) than to the rhamnosyltransferases AraGT (29% identiy, 38% similarity) from S. echinatus (38) and SpnG (29% identity, 39% similarity) from Saccharopolyspora spinosa (5). Luzhetskyy et al (39) proposed that similarity-based relationships of glycosyltransferases are better understood with consideration of the aglycone substrate than of the sugar donor.…”

“…Rhamnosyltransferases have been frequently exploited for the structural diversification of glycosylated antibiotics (4,15,38,44), as they often exhibit high flexibility towards the donor substrate. However, heterologous expression of cosmid cpzLK09 in S. fradiae A0, which produces the deoxysugars D-olivose and L-rhodinose, and Streptomyces sp.…”

“…Usually, genes involved in the formation of a bacterial secondary metabolite are clustered together. However, biosynthetic gene clusters of rhamnosylated compounds often lack the genes for L-rhamnose formation (9,16,38,54). In the case of elloramycin (48) and spinosyn (41), these genes were identified elsewhere in the genome.…”

“…1) and therefore only caprazamycin aglycones were produced by the heterologous host, Streptomyces coelicolor M512/cpzLK09 (31). The absence of genes for the formation of the L-rhamnose moiety within the corresponding biosynthetic gene cluster has previously been reported for aranciamycin (38), steffimycin (16), spinosyn (55), and elloramycin (9). Since potential genes for O methylation (cpz28 to cpz30) and a glycosyltransferase (cpz31) are encoded in the caprazamycin gene cluster, we speculated that only four genes are missing for successful heterologous production of intact caprazamycins.…”

Antibióticos: importância terapêutica e perspectivas para a descoberta e desenvolvimento de novos agentes

“…Apesar da friulimicina B apresentar similaridade química com a daptomicina, ela é o primeiro membro da nova classe de antibióticos lipopeptídicos bactericidas que parece interferir na inibição da formação do lipídeo I, um precursor na síntese de peptideoglicanos da parede celular. 28 Apesar dos nucleosídeos simples possuírem mecanismos de ação já descritos, agindo especialmente como inibidores da síntese de ácidos nucleicos, os derivados acil-e glicosil-nucleosídicos possuem mecanismos de ação mais diferenciados e complexos. Por exemplo, a puromicina e nucleocidina podem agir como inibidores da síntese de ácidos nucleicos ou proteínas ou síntese de glicanos, enquanto as nicomicinas inibem a síntese de quitina da parede celular fúngica.…”