Neutral Lipid Content in Lipid Droplets: Potential Biomarker of Cordycepin Accumulation in Cordycepin-Producing Fungi

• Main Authors: Peng Qin, ZhiYe Wang, DengXue Lu, HongMei Kang, Guang Li, Rui Guo, YuHui Zhao, RongBing Han, Bing Ji, Yang Zeng
• Format: Article
• Language: English
• Published: MDPI AG 2019-09-01
• Series: Molecules
• Subjects: lipid droplets; Nile red; cordycepin; <i>Cordyceps militaris</i>; mutants
• Online Access: https://www.mdpi.com/1420-3049/24/18/3363

To clarify the relationship between neutral lipid content and cordycepin accumulation in <i>Cordyceps</i> <i>militaris</i>, mutants were generated from mixed spores of two <i>C. militaris</i> strains with varying cordycepin-producing capacities. Fifteen stable mutants producing from 0.001 to 2.363 mg/mL cordycepin were finally selected. The relative fluorescence intensities of the 15 mutants, two <i>C. militaris</i> strains and an <i>Aspergillus nidulans</i> strain at different concentrations of lyophilized mycelium powder were then investigated using the Nile red method. The mutant CM1-1-1 with the highest relative fluorescence intensity among the eighteen strains was selected for optimizing the Nile red method. Relative fluorescence intensity was linearly correlated with cordycepin concentration in liquid broth (R² = 0.9514) and in lyophilized mycelium powder (R² = 0.9378) for the 18 cordycepin-producing strains under identical culture conditions and with cordycepin concentration in liquid broth (R² = 0.9727) and in lyophilized mycelium powder (R² = 0.9613) for CM1-1-1 under eight different sets of conditions. In addition, the cordycepin content in lyophilized mycelium powder measured by the Nile red method was linearly correlated with that determined by an HPLC method (R² = 0.9627). In conclusion, neutral lipids in lipid droplets are required during cordycepin accumulation; these neutral lipids are potential biomarkers of cordycepin biosynthesis.