Identification and expression of genes involved in biosynthesis of L-oleandrose and its intermediate L-olivose in the oleandomycin producer Streptomyces antibioticus.
A 9.8-kb DNA region from the oleandomycin gene cluster in Streptomyces antibioticus was cloned. Sequence analysis revealed the presence of 8 open reading frames encoding different enzyme activities involved in the biosynthesis of one of the two 2, 6-deoxysugars attached to the oleandomycin aglycone: L-oleandrose (the oleW, oleV, oleL, and oleU genes) and D-desosamine (the oleNI and oleT genes), or of both (the oleS and oleE genes). A Streptomyces albus strain harboring the oleG2 glycosyltransferase gene integrated into the chromosome was constructed. This strain was transformed with two different plasmid constructs (pOLV and pOLE) containing a set of genes proposed to be required for the biosynthesis of dTDP-L-olivose and dTDP-L-oleandrose, respectively. Incubation of these recombinant strains with the erythromycin aglycon (erythronolide B) gave rise to two new glycosylated compounds, identified as L-3-O-olivosyl- and L-3-O-oleandrosyl-erythronolide B, indicating that pOLV and pOLE encode all enzyme activities required for the biosynthesis of these two 2,6-dideoxysugars. A pathway is proposed for the biosynthesis of these two deoxysugars in S. antibioticus.
Aguirrezabalaga
I
I
Departamento de Biología Funcional e Instituto Universitario de Oncología de Asturias (IUOPA), Universidad de Oviedo, 33006 Oviedo, Spain.
Olano
C
C
Allende
N
N
Rodriguez
L
L
Braña
A F
AF
Méndez
C
C
Salas
J A
JA
D000595:Amino Acid Sequence; D000606:Amino Sugars / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003837:Deoxy Sugars / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial; D016695:Glycosyltransferases / Q000096:biosynthesis / Q000235:genetics; D006601:Hexoses / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009827:Oleandomycin / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D013303:Streptomyces antibioticus / Q000235:genetics* / Q000378:metabolism
D000606:Amino Sugars; D001426:Bacterial Proteins; D003837:Deoxy Sugars; D006601:Hexoses; C110125:desosamine; C035441:oleandrose; C407877:olivose; D016695:Glycosyltransferases; D009827:Oleandomycin