Identification of stsC, the gene encoding the L-glutamine:scyllo-inosose aminotransferase from streptomycin-producing Streptomycetes.
Eight new genes, strO-stsABCDEFG, were identified by sequencing DNA in the gene cluster that encodes proteins for streptomycin production of Streptomyces griseus N2-3-11. The StsA (calculated molecular mass 43.5 kDa) and StsC (45.5 kDa) proteins - together with another gene product, StrS (39.8 kDa), encoded in another operon of the same gene cluster - show significant sequence identity and are members of a new class of pyridoxal-phosphate-dependent aminotransferases that have been observed mainly in the biosynthetic pathways for secondary metabolites. The aminotransferase activity was demonstrated for the first time by identification of the overproduced and purified StsC protein as the L-glutamine:scyllo-inosose aminotransferase, which catalyzes the first amino transfer in the biosynthesis of the streptidine subunit of streptomycin. The stsC and stsA genes each hybridized specifically to distinct fragments in the genomic DNA of most actinomycetes tested that produce diaminocyclitolaminoglycosides. In contrast, only stsC, but not stsA, hybridized to the DNA of Streptomyces hygroscopicus ssp. glebosus, which produces the monoaminocyclitol antibiotic bluensomycin; this suggests that both genes are specifically used in the first and second steps of the cyclitol transamination reactions. Sequence comparison studies performed with the deduced polypeptides of the genes adjacent to stsC suggest that the enzymes encoded by some of these genes [strO (putative phosphatase gene), stsB (putative oxidoreductase gene), and stsE (putative phosphotransferase gene)] also could be involved in (di-)aminocyclitol synthesis.
Ahlert
J
J
Bergische Universität GH, FB 9, Department of Chemical Microbiology, Gauss-Str. 20, D-42097 Wuppertal, Germany.
Distler
J
J
Mansouri
K
K
Piepersberg
W
W
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004096:Dihydrostreptomycin Sulfate / Q000031:analogs & derivatives / Q000378:metabolism; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D009876:Operon; D011994:Recombinant Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D013307:Streptomycin / Q000096:biosynthesis*; D000637:Transaminases / Q000096:biosynthesis / Q000235:genetics*
D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D000637:Transaminases; C020657:glutamine-scyllo-inosose aminotransferase; C006786:bluensomycin; D004096:Dihydrostreptomycin Sulfate; D013307:Streptomycin