NMR and GC-MS Based Metabolic Profiling and Free-Radical Scavenging Activities of Cordyceps pruinosa Mycelia Cultivated under Different Media and Light Conditions

Abstract

Variation of metabolic profiles in Cordyceps pruinosa mycelia cultivated under various media and light conditions was investigated using 1H nuclear magnetic resonance (NMR) analysis and gas chromatography mass spectrometry (GC-MS) with multivariate statistical analysis. A total of 71 metabolites were identified (5 alcohols, 21 amino acids, 15 organic acids, 4 purines, 3 pyrimidines, 7 sugars, 11 fatty acids, and 5 other metabolites) by NMR and GC-MS analysis. The mycelia grown in nitrogen media and under dark conditions showed the lowest growth and ergosterol levels, essential to a functional fungal cell membrane; these mycelia, however, had the highest levels of putrescine, which is involved in abiotic stress tolerance. In contrast, mycelia cultivated in sabouraud dextrose agar with yeast extract (SDAY) media and under light conditions contained relatively higher levels of fatty acids, including valeric acid, stearic acid, lignoceric acid, myristic acid, oleic acid, palmitoleic acid, hepadecenoic acid, and linoleic acid. These mycelia also had the highest phenolic content and antioxidant activity, and did not exhibit growth retardation due to enhanced asexual development caused by higher levels of linoleic acid. Therefore, we suggested that a light-enriched environment with SDAY media was more optimal than dark condition for cultivation of C. pruinosa mycelia as biopharmaceutical or nutraceutical resources.

Metabolomic profile and nucleoside composition of Cordyceps nidus sp. nov. (Cordycipitaceae): A new source of active compounds

“…Methanol and n-hexane extractions were performed as it was described for C . pruinosa with some modifications [ 8 ]. Briefly, the extraction consisted on macerating 100 mg of the fungal material and adding 2 mL of solvent.…”

“…The total phenolic content (TPC) of methanol fractions was measured using the modified version of the Folin-Ciocalteu method [ 8 ]. The results were estimated using a calibration curve of gallic acid (Sigma-Aldrich, St. Louis, MO, USA) at a concentration ranging from 100 to 500 ppm; results are expressed as mg of gallic acid equivalents (GAE) per g of dried extract.…”

“…have been mainly attributed to nucleosides, glycosylamines involved in the regulation and modulation of several biological processes and their derivatives [ 10 – 11 ]. These may be extracted from fruiting bodies or mycelia and are currently studied as therapeutic agents [ 7 – 8 , 10 , 12 – 13 ]. Nucleosides can be differentially induced by intrinsic or environmental factors, such as the developmental stage of the fungus, light exposure and wavelength, the composition of the culture media, among other factors [ 7 – 8 ], [ 12 – 15 ].…”

“…These may be extracted from fruiting bodies or mycelia and are currently studied as therapeutic agents [ 7 – 8 , 10 , 12 – 13 ]. Nucleosides can be differentially induced by intrinsic or environmental factors, such as the developmental stage of the fungus, light exposure and wavelength, the composition of the culture media, among other factors [ 7 – 8 ], [ 12 – 15 ]. Traditionally, the nucleosides cordycepin (3’-deoxyadenosine), adenosine, inosine, guanosine, and uridine have been used as markers to assess the quality of natural and cultured species of Cordyceps [ 4 , 16 – 17 ].…”

Metabolomic profiling of the phytomedicinal constituents of Carica papaya L. leaves and seeds by 1H NMR spectroscopy and multivariate statistical analysis

“…DPPH and ABTS assays have been previously used to estimate the antioxidant properties of Italian saffron Crocus sativus L. [24], Sideritis italica extracts [25] and the antioxidant properties of Lavandula angustifolia Miller essential oil [26]. Total phenolic and flavonoid content have been previously calculated and related to plant stress in response to tetracycline accumulation [27], in Cordyceps pruinosa mycelia cultivated under different media and light conditions [28] and in Saraca Asoka flowers [29] and African medicinal plants [30]. This is the first systematic study of the metabolic profiling of papaya leaves and seeds using a combined NMR and multivariate statistical approach, and exemplifies the utility of NMR-based approaches in plant metabolomics.…”