Functional identification of the gene encoding the enzyme involved in mannosylation in ramoplanin biosynthesis in Actinoplanes sp

Ramoplanin is a lipopeptide antibiotic active against multi-drug-resistant, Gram-positive pathogens. Structurally, it contains a di-mannose moiety attached to the peptide core at Hpg¹¹. The biosynthetic gene cluster of ramoplanin has already been reported and the assembly of the depsipeptide has bee...

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Bibliographic Details
Published in:Biotechnology letters 2013-09, Vol.35 (9), p.1501-1508
Main Authors: Chen, Jun-Sheng, Wang, Yuan-Xi, Shao, Lei, Pan, Hai-Xue, Li, Ji-An, Lin, Hui-Min, Dong, Xiao-Jing, Chen, Dai-Jie
Format: Article
Language:English
Subjects:
Actinoplanes
Anti-Bacterial Agents - biosynthesis
Antibiotics
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biosynthesis
Biotechnology
Clusters
Combinatorial analysis
Depsipeptides - biosynthesis
Encoding
Enzymes
Exploitation
Gene Knockout Techniques
Genes
Genetic Complementation Test
Homologous Recombination
Life Sciences
Mannose - metabolism
Mannosyltransferases - genetics
Mannosyltransferases - metabolism
Microbial Sensitivity Tests
Microbiology
Micromonosporaceae - enzymology
Micromonosporaceae - genetics
mutants
Open Reading Frames
Original Research Paper
pathogens
Peptides
Pharmacology
sequence analysis
Sequence Analysis, DNA
Staphylococcus aureus - drug effects
sugars
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Summary:Ramoplanin is a lipopeptide antibiotic active against multi-drug-resistant, Gram-positive pathogens. Structurally, it contains a di-mannose moiety attached to the peptide core at Hpg¹¹. The biosynthetic gene cluster of ramoplanin has already been reported and the assembly of the depsipeptide has been elucidated but the mechanism of transferring sugar moiety to the peptide core remains unclear. Sequence analysis of the biosynthetic gene cluster indicated ramo-orf29 was a mannosyltransferase candidate. To investigate the involvement of ramo-orf29 in ramoplanin biosynthesis, gene inactivation and complementation have been conducted in Actinoplanes sp. ATCC 33076 by homologous recombination. Metabolite analysis revealed that the ramo-orf29 inactivated mutant produced no ramoplanin but the ramoplanin aglycone. Thus, ramo-orf29 codes for the mannosyltransferase in the ramoplanin biosynthesis pathway. This lays the foundation for further exploitation of the ramoplanin mannosyltransferase and aglycone in combinatorial biosynthesis.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-013-1233-3