Isolation of Lactobacillus salivarius from Children and Purification of Bacteriocin to Inhibition Cancer Cell in Vitro

Waleed K. M. Al-Tememy, Ibraem A. Al-Ani, Mohammed Q. Al-Ani, Nadia T. Al-Alusi, Salmah Ismail

Abstract


Bacteria being used to make anticancer agents could provide an extra source of lead compounds for the pharmaceutical industry.  Bacterium Lactobacillus salivarius produce compounds that selectively inhibit growth of human cancer cells Lactobacillus salivarius naturally produces a compound called Bacteriocins.  Bacteriocins are bacterial proteins produced to prevent the growth of competing microorganisms in a particular biological niche and we can use it as antineoplastic. The aim of this study was to isolate bacteriocin produced by lactic acid bacteria. A preparation of bacteriocin from a strain Lactobacillus salivarius has long been shown to have antineoplastic activity against a variety of human tumor and animal tumor cell lines in vitro. A total of 60 LAB  were isolated from children stool 45 isolate showed a clear antimicrobial activity against indicator strain Streptococcus aureus and by used sodium phosphate buffer (pH8) from an 80% ammonium sulfate precipitate. The inhibition  activity was determent by well diffusion assay method technique, Bacteriocin purification processes were carried out by using ion-exchange (Trisacryl SP) and gel filtration chromatography (Sephacryl – S300). The apparent molecular mass of partially purified bacteriocin was 15. 848 kDa,  Cell Culture was maintained in RPMI 1640 medium supplemented with 10% (vol/vol) fetal calf serum,  Cytotoxicity of bacteriocin was assessed on human cell line (RD) and animal cell line (MDCK) cell viability after incubation for 48 h in medium containing 500AU/ml (1.15 mg/ml). Both cell types used in this study were sensitive to bacteriocin and the bacteriocin appeared to inhibit proliferation of tumor cell line. The animal cell line was more sensitivity than human cell line.

Keywords


Lactobacillus salivarius; Bacteriocins; ion-exchange; tumor cell line

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DOI: http://dx.doi.org/10.18517/ijaseit.1.1.24

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Published by INSIGHT - Indonesian Society for Knowledge and Human Development