Sunday, August 10, 2008
P79

Application of Biolog FF MicroPlate for substrate utilization and metabolite profiling of closely related fungi

Maya P. Singh, Chemical Technologies, Chemical and Screening Sciences, Wyeth Research, 401 N. Middletown Road, Pearl River, NY 10965

Fungi are ubiquitous in nature and possess unique biochemical pathways to assimilate available nutrients and produce unique secondary metabolites, some of which are well known pharmaceuticals (penicillins, cyclosporine and statins). Since majority of the anticipated fungal cultures are believed to be unexploited screening of fungi for new chemical entities continues to be an important element in the microbial-based drug discovery paradigm. Rapid techniques for culture dereplication and selection are helpful in creating a diverse culture library for such programs. The Biolog FF MicroPlate was recently introduced for rapid identification and characterization of filamentous fungi. The FF MicroPlate contains 95 discrete substrates that are utilized differently by different species leading into distinct substrate utilization and growth fingerprints. We used the FF MicroPlate to create fingerprints for taxonomically (morphological and DNA based) related fungal cultures and compared their secondary metabolite profiles. Fingerprints were created with the Spotfire visualization software using the growth absorbance data collected on a Victor2 plate reader, and metabolite profiles were obtained by LCMS analyses of the culture extracts. Extract from each well was also tested for bioactivity against a suitable panel of bacteria and fungi. In one case only a limited number of substrates appeared to be utilized by the culture, and presence of the target compound was confirmed by antifungal activity and corresponding LCMS peak in the extracts. The substrate utilization fingerprints were also helpful in selecting media components for the secondary metabolite production optimization studies for fungal cultures.

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