Abstract
Libraries of microorganisms have been a cornerstone of drug discovery efforts since the mid-1950s, but strain duplication in some libraries has resulted in unwanted natural product redundancy. In the current study, we implemented a workflow that minimizes both the natural product overlap and the total number of bacterial isolates in a library. Using a collection expedition to Iceland as an example, we purified every distinct bacterial colony off isolation plates derived from 86 environmental samples. We employed our mass spectrometry (MS)-based IDBac workflow on these isolates to form groups of taxa based on protein MS fingerprints (3-15 kDa) and further distinguished taxa subgroups based on their degree of overlap within corresponding natural product spectra (0.2-2 kDa). This informed the decision to create a library of 301 isolates spanning 54 genera. This process required only 25 h of data acquisition and 2 h of analysis. In a separate experiment, we reduced the size of an existing library based on the degree of metabolic overlap observed in natural product MS spectra of bacterial colonies (from 833 to 233 isolates, a 72.0% size reduction). Overall, our pipeline allows for a significant reduction in costs associated with library generation and minimizes natural product redundancy entering into downstream biological screening efforts.
Original language | English |
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Pages (from-to) | 2167-2173 |
Number of pages | 7 |
Journal | Journal of Natural Products |
Volume | 82 |
Issue number | 8 |
DOIs | |
Publication status | Published - 23 Aug 2019 |
Bibliographical note
Funding Information:The authors would like to acknowledge E. Bogason and his team at the Strytan Dive Center and E. Einarsdottir for assistance in sample collection, and M. Sadek, Y. Gao, and C. Murphy for assistance with microbial cultivation. Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R01GM125943 (B.T.M. and L.M.S.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This work was also supported by Icelandic Research Fund Grant 152336-051 (B.T.M. and S.O.) and UIC startup funds (L.M.S.).
Publisher Copyright:
Copyright © 2019 American Chemical Society and American Society of Pharmacognosy.