Optimization parameters for the production of dimer of epicatechin from an endophytic fungus Curvularia australiensis FC2AP using response surface methodology (RSM).

The search for natural therapeutic agents has intensified due to their potential to treat various diseases. Bioactive secondary metabolites from endophytes offer high therapeutic profiles and can be mass-produced after optimizing medium parameters and purification. This investigation aimed to maximize crude pigmented secondary metabolite (CPSM) production from Curvularia australiensis FC2AP by optimizing fermentation conditions statistically. The endophytic fungus produced a maximum yield of 8.81 UL/g from biomass using Sabouraud's Dextrose Broth. After screening essential factors, the Plackett-Burman design was used for factorial optimization, and the Box Behnken design was employed to investigate three significant factors. The final CPSM yield was 12.3 UL/g, approximately 4-fold higher than the preliminary growth medium. Chromatographic purification using a gradient solvent system resulted in six fractions, with the fourth fraction demonstrating the highest bioactivity profile. Structural characterization confirmed this fraction to be a dimer of epicatechin, which has anti-cancer properties, as confirmed through in vivo studies on Sprague Dawley rats. This is the first report of a epicatechin dimer produced from C. australiensis.

Evaluation of Dimer of Epicatechin from an Endophytic Fungus Curvularia australiensis FC2AP on Acute Toxicity Levels, Anti-Inflammatory and Anti-Cervical Cancer Activity in Animal Models.

Cervical cancer, as the most frequent cancer in women globally and accounts almost 14% in India. It can be prevented or treated with vaccines, radiation, chemotherapy, and brachytherapy. The chemotherapeutic agents cause adverse post effects by the destruction of the neighboring normal cells or altering the properties of the cells. In order to reduce the severity of the side effects caused by the chemically synthesized therapeutic agents, the current research developed an anti-cancer agent dimer of epicatechin (DoE), a natural bioactive secondary metabolite (BSM) mediated from an endophytic fungus Curvularia australiensis FC2AP. The investigation has initiated with the evaluation of inhibiting the angiogenesis which is a main activity in metastasis, and it was assessed through Hen's Egg Test on Chorio Allantoic Membrane (HET-CAM) test; the BSM inhibited the growth of blood vessels in the developing chick embryo. Further the DoE was evaluated for its acute toxicity levels in albino mice, whereas the survival dose was found to be 1250 mg/kg and the lethal dose was 1500 mg/kg body weight of albino mice; hematological, biochemical, and histopathological analyses were assessed. The anti-inflammatory responses of the DoE were evaluated in carrageenan induced Wistar rats and the reduction of inflammation occurred in a dose-dependent manner. By fixing the effective dose for anti-inflammation analysis, the DoE was taken for the anti-cervical cancer analysis in benzo (a) pyrene induced female Sprague-Dawley rats for 60 days trial. After the stipulated days, the rats were taken for hematological antioxidants, lipid peroxidation (LPO), member bound enzymes, cervical histopathological and carcinogenic markers analyses. The results specified that the DoE has the capability of reducing the tumor in an efficient way. This is the first report of flavonoid-DoE production from an endophytic fungus C. australiensis has the anticancer potentiality and it can be stated as anti-cancer drug.