Molecular cloning and characterization of a 2-deoxystreptamine biosynthetic gene cluster in gentamicin-producing Micromonospora echinospora ATCC15835.
The organization of the 2-deoxystreptamine (DOS) biosynthetic gene cluster of Micromonospora echinospora has been determined. Sequencing of a 14.04 kb-region revealed twelve open reading frames (ORFs): four putative DOS biosynthetic genes (gtmA, B, C, and D), five amino sugars biosynthetic genes (gtmE, G, H, I, and gacB), two aminoglycoside resistance genes (gtmF and J) as well as a hypothetical ORF (gacA). One of the putative DOS biosynthetic genes, gtmA, was expressed in Escherichia coli, and the purified protein was shown to convert glucose-6-phosphate (G-6-P) to 2-deoxy-scyllo-inosose (DOI), a key step in DOS biosynthesis. In addition gtmJ was expressed in Streptomyces lividans and shown to confer gentamicin resistance. Thus gtmA and gtmJ are implicated in the biosynthesis of gentamicin and in resistance to it, respectively.
Kharel
Madan K
MK
Institute of Biomolecule Reconstruction, Sun Moon University, Asan 336-708, Korea.
Basnet
Devi B
DB
Lee
Hei Chan
HC
Liou
Kwangkyoung
K
Moon
Young Ho
YH
Kim
Jae-Jong
JJ
Woo
Jin Suk
JS
Sohng
Jae Kyung
JK
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism; D003001:Cloning, Molecular; D024881:Drug Resistance, Bacterial; D005839:Gentamicins / Q000096:biosynthesis* / Q000737:chemistry; D019298:Glucose-6-Phosphate / Q000378:metabolism; D006595:Hexosamines / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008847:Micromonospora / Q000235:genetics* / Q000378:metabolism; D015394:Molecular Structure; D005810:Multigene Family; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D006595:Hexosamines; D019298:Glucose-6-Phosphate; C007265:2-deoxystreptamine