Characterization of RbmD (glycosyltransferase in ribostamycin gene cluster) through neomycin production reconstituted from the engineered Streptomyces fradiae BS1.
Amino acid homology analysis predicted that rbmD, a putative glycosyltransferase from Streptomyces ribosidificus ATCC 21294, has the highest homology with neoD in neomycin biosynthesis. S. fradiae BS1, in which the production of neomycin was abolished, was generated by disruption of the neoD gene in the neomycin producer S. fradiae. The restoration of neomycin by self complementation suggested that there was no polar effect in the mutant. In addition, S. fradiae BS6 was created with complementation by rbmD in S. fradiae BS1, and secondary metabolite analysis by ESI/MS, LC/MS and MS/MS showed the restoration of neomycin production in S. fradiae BS6. These gene inactivation and complementation studies suggested that, like neoD, rbmD functions as a 2-N-acetlyglucosaminyltransferase and demonstrated the potential for the generation of novel aminoglycoside antibiotics using glycosyltransferases in vivo.
Nepal
Keshav Kumar
KK
Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, Sun Moon University, Asan, 336-708, Korea.
Oh
Tae-Jin
TJ
Subba
Bimala
B
Yoo
Jin Cheol
JC
Sohng
Jae Kyung
JK
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005818:Genetic Engineering; D016695:Glycosyltransferases / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D005810:Multigene Family; D009154:Mutation / Q000235:genetics; D009355:Neomycin / Q000096:biosynthesis*; D012271:Ribostamycin / Q000737:chemistry / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D000900:Anti-Bacterial Agents; D012271:Ribostamycin; D016695:Glycosyltransferases; D009355:Neomycin