Purification of The Antitumor Active Metabolite from Streptomyces xylophagus Ha.Ph-7 Crude Extract Isolated from Hamam Pharaoh Hot Spring

Abstract

Aim: This study aimed at production, extraction, purification and characterization of active antitumor compound from marine actinomycete secondary metabolites crude extract that was selected as most potent crude secondary metabolites from the preliminary screening of total fifty one extract. Material and methods: These tumor cell lines were breast cancer (MCF-7) (ATCC HTB-22) and Hepato-cellular carcinoma (HepG2) (ATCC 77400), colon cancer (Caco) (ATCC HTB-37) and cervix carcinoma(HeLa) (ATCC CRL-13011) cells. Purification process was done using silica gel column chromatography while purity was detected using thin layer chromatography (TLC). The active fraction was detected using morphological changes (cytotoxicity) by microscopic examination and anti-proliferative activity using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Result: The result indicated that there is only one fraction responsible for antitumor activity. This active fraction has cytotoxic and anti-proliferative activity against HepG2, MCF7, Caco and HELA cells with IC50=24.5, 20.1, 27.6 and 17.7µg/ml respectively. The analysis of physico-chemical, elemental and spectroscopic analysis (UV, IR, H.NMR, Mass spectroscopy) indicated that; the active compound has the nature of anthracycline compound. Conclusion: This purified compound has promising broad spectrum cytotoxicity and anti-proliferative activity in an in vitro system. The next research step shall include different optimization parameter that maximize productivity with long-term goal; discovery of an antitumor drug from actinomycetes native to Egyptian hot springshabitat.