Production Optimization and In Vitro Evaluation of Anti-Proliferative, Anti-Oxidant and Anti-Inflammatory Potential of the Antibacterial Peptide MAFP9

Abstract

\n Peptides are important natural bioactive agents obtained from various sources including microbes, plants and animals. The present study involved the optimization and physicochemical characterization of an antibacterial peptide MFAP9 produced by a marine fungus Aspergillus fumigatus BTMF9 and its vitro evaluation of antiproliferative, antioxidant and anti-inflammatory potential. Antiproliferative activity was performed using A549 lung carcinoma and normal L929 fibroblast cell lines employing MTT assay method. Antioxidant activity was assessed using DPPH assay and Cox2 inhibition assay of peptide treated LPS stimulated THP1 cell lines was used for anti-inflammatory studies. The results of in vitro antiproliferative activity assessment showed that MFAP9 was cytotoxic with an IC50 value of 29 µg/mL towards A549 cells. Fluorescence microscopic analysis revealed apoptotic features such as chromatin condensation and alterations in the size and the shape of cells after treatment with MFAP9 at a concentration 50 µg/mL. The DPPH radical scavenging activity of MFAP9 exhibits reasonably higher scavenging activity compared to that of standard antioxidant at same concentration level. Moreover, MFAP9 showed significant anti-inflammatory activities. Time course experiment was conducted with the Czapek- Dox medium and optimized the rate of MFAP9 production. The study on various physico-chemical parameters on the stability revealed that the maximum active temperature of MAFP9 is 50oC, stable in basic pH and 1 mM concentration of Ni2+, Cd2+ and Mg2+ promotes the peptide activity. Hopefully, by employing the advancements in proteomics, bioinformatics, and modification strategies, MFAP9 could emerge as novel promising therapeutic drug in future for various clinical applications.