Metabolites were extracted from 5 mg freeze-dried fungal mycelia and dried in a speed vacuum as described previously (28 (link)). Extracts were reconstituted in 50 μl of deuterated sodium phosphate buffer (100 mM, pH 7.0) containing 0.5 mM trimethylsilylpropanoic acid (TMSP), 3 mM sodium azide, and 100% D2O. Each sample was sonicated for 10 min and vortexed briefly, before a volume of 35 μl was transferred into 1.7-mm nuclear magnetic resonance (NMR) tubes.
Spectra were acquired on a Bruker 600 MHz spectrometer equipped with a TCI 1.7-mm z-PFG cryogenic probe and a Bruker SampleJet autosampler. One-dimensional (1D) 1H NMR spectra and 2D 1H-13C HSQC (heteronuclear single quantum coherence) spectra were recorded and analyzed for each sample as previously described (63 (link)).
Spectra were acquired on a Bruker 600 MHz spectrometer equipped with a TCI 1.7-mm z-PFG cryogenic probe and a Bruker SampleJet autosampler. One-dimensional (1D) 1H NMR spectra and 2D 1H-13C HSQC (heteronuclear single quantum coherence) spectra were recorded and analyzed for each sample as previously described (63 (link)).
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