Garcia Ronald O, Krug Daniel, Müller Rolf
Department of Pharmaceutical Biotechnology, Saarland University, Saarbrücken, Germany.
Methods Enzymol. 2009;458:59-91. doi: 10.1016/S0076-6879(09)04803-4.
Myxobacteria produce a range of structurally novel natural products which exhibit unusual or unique modes of action, attracting significant interest from both the academic and drug discovery communities. Efforts to discover new strains with the potential to biosynthesize novel molecules have revealed that myxobacterial diversity and natural products are far from exhausted. We describe here a general, nonselective approach to unearth further myxobacterial strains, in order to mine them for compounds with potential as medicines. Sample collection from locations world-wide has shown that environments which exhibit significant biological complexity yield the highest probability of isolating novel myxobacterial strains. Here, we illustrate the details of simple and efficient strain purification techniques, which lead systematically to the identification of new and promising myxobacteria. Compound identification is then facilitated by molecular biological approaches, coupled with sophisticated high resolution mass spectrometry, statistical analysis, and bioassays.
粘细菌能产生一系列结构新颖的天然产物,这些产物具有不同寻常或独特的作用方式,引起了学术界和药物研发界的极大兴趣。为发现具有生物合成新分子潜力的新菌株所做的努力表明,粘细菌的多样性和天然产物远未穷尽。我们在此描述一种通用的、非选择性的方法来挖掘更多的粘细菌菌株,以便从中筛选出具有药用潜力的化合物。从全球各地采集样本表明,具有显著生物复杂性的环境分离出新的粘细菌菌株的可能性最高。在此,我们阐述简单高效的菌株纯化技术细节,这些技术能系统地引导我们鉴定出新的、有前景的粘细菌。然后,通过分子生物学方法,结合先进的高分辨率质谱、统计分析和生物测定,来促进化合物的鉴定。