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qNMR for profiling the production of fungal secondary metabolites
Authors:Wilson C. Brooks  Noemi D. Paguigan  Huzefa A. Raja  Franklin J. Moy  Nadja B. Cech  Cedric J. Pearce  Nicholas H. Oberlies
Affiliation:1. Department of Chemistry and Biochemistry, University of North Carolina at Greensboro, Greensboro, NC, USA;2. Mycosynthetix, Inc, Hillsborough, NC, USA
Abstract:Analysis of complex mixtures is a common challenge in natural products research. Quantitative nuclear magnetic resonance spectroscopy offers analysis of complex mixtures at early stages and with benefits that are orthogonal to more common methods of quantitation, including ultraviolet absorption spectroscopy and mass spectrometry. Several experiments were conducted to construct a methodology for use in analysis of extracts of fungal cultures. A broadly applicable method was sought for analysis of both pure and complex samples through use of an externally calibrated method. This method has the benefit of not contaminating valuable samples with the calibrant, and it passed scrutiny for line fitting and reproducibility. The method was implemented to measure the yield of griseofulvin and dechlorogriseofulvin from three fungal isolates. An isolate of Xylaria cubensis (coded MSX48662) was found to biosynthesize griseofulvin in the greatest yield, 149 ± 8 mg per fermentation, and was selected for further supply experiments. Copyright © 2016 John Wiley & Sons, Ltd.
Keywords:NMR  1H NMR  qNMR  griseofulvin  fungi  secondary metabolites
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