Taxonomic group: bacteria / Actinobacteria
(Phylum: Actinobacteria)
Associated disease: infection due to Actinomadura [ICD11:
XN8P7 
]
NCBI PubMed ID: 25878261Publication DOI: 10.1099/mic.0.000095Journal NLM ID: 0376646Publisher: Washington, DC: Kluwer Academic/Plenum Publishers
Correspondence: Jason.micklefield
manchester.ac.uk
Institutions: School of Chemistry and Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK
The lipopeptides ramoplanin from Actinoplanes sp. ATCC 33076 and enduracidin produced by Streptomyces fungicidicus are effective antibiotics against a number of drug-resistant Gram-positive pathogens. While these two antibiotics share a similar cyclic peptide structure, comprising 17 amino acids with an N-terminal fatty acid side chain, ramoplanin has a di-mannose moiety that enduracidin lacks. The mannosyl substituents of ramoplanin enhance aqueous solubility, which was important in the development of ramoplanin as a potential treatment for Clostridium difficile infections. In this study we have determined the function of the putative mannosyltransferase encoded by ram29 from the ramoplanin biosynthetic gene cluster. Bioinformatics revealed that Ram29 is an integral membrane protein with a putative DxD motif that is suggested to bind to, and activate, a polyprenyl phosphomannose donor and an extracytoplasmic C-terminal domain that is predicted to bind the ramoplanin aglycone acceptor. The ram29 gene was cloned into the tetracycline inducible plasmid pMS17 and integrated into the genome of the enduracidin producer S. fungicidicus. Induction of ram29 expression in S. fungicidicus resulted in the production of monomannosylated enduracidin derivatives, which are not present in the WT strain. Tandem MS analysis showed that mannosylation occurs on the Hpg11 residue of enduracidin. In addition to confirming the function of Ram29, these findings demonstrate how the less common, membrane-associated, polyprenyl phosphosugar-dependent glycosyltransferases can be used in natural product glycodiversification. Such a strategy may be valuable in future biosynthetic engineering approaches aimed at improving the physico-chemical and biological properties of bioactive secondary metabolites including antibiotics
biosynthesis, antibiotics, nonribosomal peptides, lipoglycopeptides, mannosylation, glycodiversification
Structure type: oligomer
Location inside paper: Fig. 1, A40926 (dalbavancin precursor)
Trivial name: A40926
Compound class: glycoside
Contained glycoepitopes: IEDB_130701,IEDB_144983,IEDB_152206,IEDB_983930,SB_44,SB_67,SB_72
Methods: PCR, DNA sequencing, DNA techniques, HPLC, UV, extraction, ESI-QTOF-MS/MS, LC-ESI-MS, cell growth, centrifugation
Related record ID(s): 47727, 47729, 47730, 47733, 47736
NCBI Taxonomy refs (TaxIDs): 93944
Show glycosyltransferases
There is only one chemically distinct structure:
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(later renamed to: Nonomuraea gerenzanensis ATCC 39727)