In vitro Antifungal, Antioxidant and Cytotoxic Activities of a Partially Purified Protein Fraction from Atlantia monophylla Linn (Rutaceae) Leaf
Purpose: To determine the in vitro antifungal and antioxidant activities of the aqueous extract and protein fraction of Atlantia monophylla Linn (Rutaceae) leaf.
Methods: Ammonium sulphate (0 – 80 %) precipitation method was used to extract protein from the leaves of A. monophylla Linn (Rutaceae). In vitro antifungal assays were performed by disc-diffusion and micro-broth dilution methods. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2), superoxide and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities were performed to evaluate in vitro antioxidant activities. Sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) was employed to assess the molecular weight of the protein fractions. Protein concentration was determined by Bradford method. The cytotoxicity of these extracts was tested on Vero cell lines.
Results: Both the aqueous extract and protein fraction (AMP III) of Atlantia monophylla leaf exhibited higher antifungal activity on Candida albicans than on Aspergillus fumigatus. AMP III fraction showed greater in vitro antioxidant activity than the aqueous extract. SDS-PAGE analyses revealed the presence of two protein bands with molecular weight approximately of 16 and 67 KDa in AMP III. Protein concentration was 240 μg/ml in the aqueous extract and 670 μg/ml in AMP III fraction. The aqueous extract and protein fraction exhibited cytotoxicity at concentrations > 100 μg/ml on Vero cells.
Conclusion: Plant-derived proteins/peptides possessing antifungal and antioxidant properties would be a good alternative preparation for the treatment of infectious diseases.
Keywords: Atlantia monophylla, Antifungal, Antioxidant, Cytotoxicity, Proteins/peptides, Vero cell lines