Isolation and characterization of the tobramycin biosynthetic gene cluster from Streptomyces tenebrarius
- 作者：Kharel ; Madan Kumar ; Basnet ; Devi Bahadur ; Lee ; Hei Chan ; Liou ; Kwangkyoung ; Woo ; Jin Suk ; et. al.
- 关键词：Aminoglycoside ; 2-Deoxy-scyllo-inosose synthase ; 2-Deoxystreptamine ; Streptomyces
- 刊名：FEMS Microbiology Letters
- 出版年：2004
- 期刊代码：34_03781097
- 类别：imm
- 出版时间：January 30, 2004
- 卷：230
- 期：2
- 页码：185-190
- 文件大小：375 K
摘要
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.