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...

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Published in:Molecules and cells 2009, 27(1), , pp.83-88
Main Authors: Nepal, Keshav Kumar (Sunmoon University, Asan, Republic of Korea), Oh, T.J. (Sunmoon University, Asan, Republic of Korea), Subba, Bimala (Sunmoon University, Asan, Republic of Korea), Yoo, J.C. (Chosun University, Gwangju, Republic of Korea), E-mail: jcyu@chosun.ac.kr, Sohng, J.K. (Sunmoon University, Asan, Republic of Korea), E-mail: sohng@sunmoon.ac.kr
Format: Article
Language:English
Subjects:
aminoglycoside
Anti-Bacterial Agents - pharmacology
Biochemistry
Biomedical and Life Sciences
Biomedicine
Biotechnology
Cell Biology
Genes, Bacterial
Genetic Complementation Test
Genetic Engineering
GLICOSILTRANSFERASAS
GLYCOSYLTRANSFERASE
GLYCOSYLTRANSFERASES
Glycosyltransferases - genetics
Life Sciences
Microbial Sensitivity Tests
Multigene Family
Mutation - genetics
NEOMICINA
NEOMYCIN
Neomycin - biosynthesis
NEOMYCINE
ribostamycin
Ribostamycin - chemistry
Ribostamycin - metabolism
Sequence Analysis, DNA
Spectrometry, Mass, Electrospray Ionization
STREPTOMYCES
Streptomyces - drug effects
Streptomyces - enzymology
Streptomyces - genetics
생물학
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Summary: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.
ISSN:1016-8478
0219-1032
DOI:10.1007/s10059-009-0008-0