Identification and purification of bioactive compounds from wild Irpex lacteus

Abstract

newline viii newlineABSTRACT newlineIrpex lacteus is a white-rot fungus and has been reported as a medicinal mushroom. It belongs to the Polyporaceae family having a tooth-like pore structure. I. lacteus have been mentioned in the traditional Chinese medicinal system for drug formulations and used as a treatment for chronic glomerulonephritis. Recent research works have shown the antimicrobial, antioxidant, and anticancer properties of I. lacteus. Various bioactive compounds isolated from I. lacteus have been demonstrated for many medicinal activities. newlineIn the present study different solvent extracts (methanol, hexane, and aqueous) of wild I. lacteus mycelial culture were evaluated for medicinal properties. The antimicrobial activity as done against Staphylococcus aureus, Bacillus subtilis Escherichia coli, Candida albicans, Klebsiella pneumonia, and also antioxidation properties by ABTS.+ decolorization assay was performed. The bioactive compounds screening was done in the I. lacteus extract by phenol and flavonoid quantification and further estimated by HPTLC and HPLC. Purification of bioactive compound from the hydro-alcoholic extract was done by Silica gel Column chromatography. Further cytotoxicity of the extract and the purified bioactive compound was analyzed by using MTT assay against lung carcinoma A549 and pancreatic cancer MIA PaCa-cell lines. Cultivation of I. lacteus fruiting bodies was also done in different substrates. The present finding demonstrated that the methanolic extract had significantly high antibacterial (S. aureus 22.3 ± 0.33 mm; Minimal Inhibitory Concentration 6.2 mg/ml) and antioxidation activity (67.27 ± 0.34 and#956;g/ml). The antifungal activity was best observed in the case of an aqueous extract with MIC 6.25 mg/ml. The phytochemical analysis disclosed the presence of major bioactive compounds viz phenol, flavonoids, alkaline, tannin, and saponins. HPTLC and HPLC estimation of the methanolic extract showed a good concentration of gallic acid, quercetin, rutin, and catechin bioactive compounds. newlineIP-1 bioactive compound was isolated from methanolic extract using column chromatography. 1H and 13C NMR spectra confirmed the IP-1 compound as and#946;-sitosterol. MTT assay showed a mild activity by the methanolic extract. I. lacteus methanolic extract showed high cancer cell death in A549 (60%) and MIA PaCa-2 newlineix newline(54.5%) at 200 and#956;g/ml. The IC50 of I. lacteus methanolic extract on A549 and Mia-paca cells were 122.6±1.07 and#956;g/ml and 154. 2±1.06 and#956;g/ml respectively. The anticancer activity of IP-1 on A549 and MIA PaCa-2 cancer cell lines exhibited potent inhibitory activities comparable to that of Doxorubicin. IP-1 has shown high cancer cell death in A549 (62.44%) and MIA PaCa-2 (64.92%) at 200 and#956;g/ml. The IC50 of IP-1 on A549 and MIA PaCa-2 cells were 116.7±4.06 and#956;g/ml and 108.9±3.07 and#956;g/ml respectively. I. lacteus fruiting bodies were cultivated on the wheat straw substrate. Spawn preparation was completed after 8-10 days at 28°C. A mature fruiting body was harvested in the wheat straw substrate after 28 days. newlineThe nutritional profile of cultivated fruiting showed the presence of protein, carbohydrate, and fiber content. Cultivated fruiting bodies showed extracellular enzyme secretion of laccase, MnP, and cellulases. The results of this study showed the medicinal potential of I. lacteus extracts and can be used in the development of a new pharmaceutical agent.