Cloning and characterization of new glycopeptide gene clusters found in an environmental DNA megalibrary.

Abstract

Glycopeptide antibiotics have long served as drugs of last resort for the treatment of antibiotic-resistant gram-positive bacterial infections. Resistance to the clinically relevant glycopeptides, vancomycin and teicoplanin, threatens to undermine the usefulness of this important class of antibiotics. DNA extracted from a geographically diverse collection of soil samples was screened by PCR for the presence of sequences related to OxyC, an oxidative coupling enzyme found in glycopeptide biosynthetic gene clusters. Every soil sample examined contained at least 1 unique OxyC gene sequence. In an attempt to access the biosynthetic gene clusters associated with these OxyC sequences, a 10,000,000-membered environmental DNA (eDNA) megalibrary was created from a single soil sample. Two unique glycopeptide gene clusters were recovered from this eDNA megalibrary. Using the teicoplanin aglycone and the 3 sulfotransferases found in one of these gene clusters, mono-, di-, and trisulfated glycopeptide congeners were produced. The high frequency with which OxyC genes were found in environmental samples indicates that soil eDNA libraries are likely to be a rewarding source of glycopeptide gene clusters. Enzymes found in these gene clusters should be useful for generating new glycopeptides analogs. Environmental DNA megalibraries, like the one constructed for this study, can provide access to many of the natural product biosynthetic gene clusters that are predicted to be present in soil microbiomes.

• Authors: Jacob J Banik, Sean F Brady
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 105 Issue 45 Pg. 17273-7 (Nov 11 2008) ISSN: 1091-6490 [Electronic] United States
• PMID: 18987322 (Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Anti-Bacterial Agents
• DNA Primers
• Glycopeptides
• Soil
• DNA
• Sulfotransferases

Topics

• Anti-Bacterial Agents
• Base Sequence
• Chromatography, High Pressure Liquid
• Cloning, Molecular
• Cluster Analysis
• DNA (genetics, isolation & purification)
• DNA Primers (genetics)
• Gene Library
• Glycopeptides (genetics)
• Molecular Sequence Data
• Molecular Structure
• Multigene Family (genetics)
• Phylogeny
• Sequence Analysis, DNA
• Soil (analysis)
• Sulfotransferases (genetics)
• Utah