Isolation and characterization of the tobramycin biosynthetic gene cluster from Streptomyces tenebrarius.
The biosynthetic gene cluster for tobramycin, a 2-deoxystreptamine-containing aminoglycoside antibiotic, was isolated from Streptomyces tenebrarius ATCC 17920. A genomic library of S. tenebrarius was constructed, and a cosmid, pST51, was isolated by the probes based on the core regions of 2-deoxy-scyllo-inosose (DOI) synthase, and L-glutamine:DOI aminotransferase and L-glutamine:scyllo-inosose aminotransferase. Sequencing of 33.9 kb revealed 24 open reading frames (ORFs) including putative tobramycin biosynthetic genes. We demonstrated that one of these ORFs, tbmA, encodes DOI synthase by in vitro enzyme assay of the purified protein. The catalytic residues of TbmA and dehydroquinate synthase were studied by homology modeling. The gene cluster found is likely to be involved in the biosynthesis of tobramycin.
Kharel
Madan Kumar
MK
Institute of Biomolecule Reconstruction, Sun Moon University, #100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, South Korea.
Basnet
Devi Bahadur
DB
Lee
Hei Chan
HC
Liou
Kwangkyoung
K
Woo
Jin Suk
JS
Kim
Byung-Gee
BG
Sohng
Jae Kyung
JK
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002076:Butirosin Sulfate / Q000096:biosynthesis; D007294:Inositol / Q000031:analogs & derivatives* / Q000378:metabolism; D008190:Lyases / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D014031:Tobramycin / Q000096:biosynthesis*
D001426:Bacterial Proteins; D007294:Inositol; D002076:Butirosin Sulfate; C075548:2-deoxyinosose; D008190:Lyases; C085631:2-deoxy-scyllo-inosose synthase; D014031:Tobramycin