Engineering deoxysugar biosynthetic pathways from antibiotic-producing microorganisms. A tool to produce novel glycosylated bioactive compounds.
A plasmid (pLN2) was generated in which genes involved in the biosynthesis of L-oleandrose in the oleandomycin producer Streptomyces antibioticus ATCC11891 were cloned. pLN2 was used to direct the biosynthesis of different deoxysugars by exchanging and/or adding genes from other antibiotic biosynthetic clusters. Transfer of the synthesized deoxysugars to the tetracenomycin C aglycon, 8-demethyl-tetracenomycin C, through the use of the "sugar flexible" glycosyltransferase ElmGT, validated the system. Several pLN2 derivatives were constructed by replacement of the oleU 4-ketoreductase gene by different 4-ketoreductase genes. Some of them, such as EryBIV and UrdR, reduced the keto group of the 4-keto intermediates, generating L-olivosyl and D-olivosyl derivatives, respectively. The system was also used to generate an L-rhamnosyl derivative (through a two-gene deletion) and an L-rhodinosyl derivative (through a combination of a gene replacement and a gene addition).
Rodríguez
Leticia
L
Departamento de Biología Funcional e Instituto Universitario de Oncología del Principado de Asturias, Universidad de Oviedo, 33006, Oviedo, Spain.
Aguirrezabalaga
Ignacio
I
Allende
Nerea
N
Braña
Alfredo F
AF
Méndez
Carmen
C
Salas
José A
JA
D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D003001:Cloning, Molecular; D003837:Deoxy Sugars / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015195:Drug Design; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D005798:Genes, Bacterial / Q000235:genetics*; D005818:Genetic Engineering; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D006601:Hexoses / Q000096:biosynthesis* / Q000737:chemistry; D010957:Plasmids / Q000235:genetics*; D013303:Streptomyces antibioticus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D003837:Deoxy Sugars; D006601:Hexoses; C035441:oleandrose; C407877:olivose; D016695:Glycosyltransferases