ISOLATION AND CHARACTERIZATION OF ANTICANCEROUS BIOACTIVE COMPOUNDS FROM MARINE BACILLUS SUBTILIS

Abstract

In the present research, soil samples were collected from Nijampatanam marine sediments and isolated bacteria were identified through 16s rRNA and mass production of bioactive metabolite was carried out by optimizing physic-chemical parameters such as temperature, incubation, NaCl concentration, carbon and nitrogen sources. Chromatographically purified crude compound was structurally identified and biological analysis like anticancer properties in both in vitro and In silico were done with MTT assay and auto dock 4.2 against human, breast cancer cell line. Out of 10 distinct isolates, isolated from Nijampatanam marine sediments, only one; MRRPK4, exhibited considerable amount of anticancer activity. Molecular sequence shows that the isolated bacterium belongs to the genus Bacillus and species subtilis sub sp inaquosorum with the genbank accession number MT421945. Mass production of biologically active compound achieved by using 60hr incubation, 35˚C temperature, pH 7 and NaCl concentration 5.0 %. 1.0 % fructose and peptone have shown excellent growth and production. Ethyl acetate serves as a suitable solvent for extraction and chromatographically purified compound shows a novel structure (3-(3- methylbutan-2-yl) octahydropyrrolo(1,2-a)pyrazine-1,4,dione) with the molecular weight of 238g/mol. Purified compound showing concentration dependent antioxidant activity. In vitro MTT assay of purified compound shows concentration dependent activity against MCF-7 cell lines with IC50 values of 12.32μg/ml. This activity was confirmed by docking of purified compound with proteins of above tested cancer cell lines (HER2 proteins) and standard drug doxorubicin yields similar results with greater binding energy, inhibition constant and greater number of amino acid interaction with hydrogen bonds.