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ZoP Cytotoxic Property of 50kDa Protein Isolated from Streptomyces A13 against A549 Lung Cancer Cells and HCT116 Healthy Hamster Cells

autorzy: MARIA LEONORA L. DELOS SANTOS, HANNAH MABEL H. BANOCNOC, JOANA ROSE NACIONAL, MARY ANGELIE KATHLEEN BADONG

Streptomyces are known to produce anticancer compounds that are effective inhibitors of some cancer cells. We would like to determine if 50 kDA protein from Streptomyces A13 have cytotoxic properties against A549 lung cancer cells.A13 were grown on CVM medium. Cells were separated by centrifugation at 3,000 rpm. Extraction of the proteins from the cells was performed through ammonium-sulfate precipitation. Protein content was measured by the Bradford Method with bovine serum albumin as the reference protein. The precipitated proteins were analysized for size by denaturing PAGE using standard procedures. The ammonium sulfate-precipitated broth (B1) sample and cell extract were used for the MTT assay performed at the General Equipment Laboratory of the University of the Philippines, Diliman.The protein concentration was highest with the B1 (446.323ug/mL) followed by the spent medium (215.700 ug/mL), cell extract (96.158ug/mL) and least in the PEI pellet (65.166ug/mL).This high protein concentration in B1 as compared to the cell extract may be explained by their extrusion from the cell. A13 cell extract and B1 were found cytotoxic against the A549 lung cancer cells having mean IC50 values of 4.37013 μg/ml and 4.54037 μg/ml respectively. The positive control, Doxorubicin had a mean IC50 value of 2.27627 μg/ ml. Identification of the A13 was carried out using the 16s rRNA full gene sequence. BLAST result of the 16s rRNA for A13 yielded 99% homology to Streptomyces costaricanus, Streptomyces padanus and other 15 Streptomyces sp. isolates.