Development of an efficient method for separation and purification of cordycepin from liquid fermentation of Cordyceps militaris and analysis of cordycepin antitumor activity.

Cordycepin (3 '-deoxyadenosine) is the main active component of Cordyceps militaris, which is a chemical marker for quality detection of Cordyceps militaris and has important medicinal development value. Existing methods for obtaining cordycepin are complex and costly. In this study, an economical and simple method for separation and purification of cordycepin from Cordyceps militaris fermentation liquid through physical crystallization was explored. First, lyophilized powdered fermentation liquid (LPFL) and pure methanol (1 g/100 mL, w/v) were mixed, and then repeatedly dissolved and crystallized until the precipitation was white. Purified product was obtained by freeze-drying the precipitate. The substance was determined to be cordycepin by high performance liquid chromatography, mass spectrometry and infrared spectroscopy, and the purity was 94.26%. Compared with the existing methods, this method is simple and low cost. In addition, the functional activity of cordycepin was determined by in vitro test. The results exhibited that cordycepin caused death and morphological changes in human colon cancer Caco-2 cells, and significantly inhibited the proliferation of Caco-2 cells, with a half-maximal inhibitory concentration (IC50) of 107.2 µg/mL. Cordycepin could induce early apoptosis of Caco-2 and caused cell cycle arrest in the G2 phase. Caco-2 cell apoptosis and cell cycle arrest showed dose dependence to cordycepin over a certain range. These results improved cordycepin purification method, provided insights into the mechanism of cordycepin in cancer inhibition, and would provide important reference for further development and clinical application of cordycepin.

In Vitro Antibacterial and Antitumor Activity of Total Triterpenoids from a Medicinal Mushroom Sanghuangporus sanghuang (Agaricomycetes) in Liquid Fermentation Culture.

We analyzed the biological activity and composition of total triterpenoids extracted from the liquid fermentation culture of the medicinal mushroom Sanghuangporus sanghuang. S. sanghuang total triterpenoid extract (STTE) had the strongest inhibitory effect on Staphylococcus aureus, and STTE at 2-fold the minimum inhibitory concentration had a significant effect on the growth of four types of bacteria and caused marked nucleic acid leakage. Scanning electron microscopy confirmed that STTE destroyed bacterial cell walls. STTE also significantly inhibited the proliferation of Caco-2 cells and disrupted their morphology. Flow cytometry revealed that STTE induced cell cycle arrest and induced early and late apoptosis of Caco-2 cells. Infrared spectroscopy, and gas chromatography-mass spectrometry resulted in the detection of a variety of triterpenoids (e.g., pentacyclic triterpenoid, lanolinane triterpenoid, and trans-squalene) as well as other small molecules (e.g., esters, acids, and aldehydes). Our findings indicate that STTE has antibacterial and antitumor activity and will enable screening of novel fungal-derived anticancer and antibacterial compounds.