Abstract

EXTRACTION AND PURIFICATION OF BACTERIOCIN FROM STREPTOCOCCUS THERMOPHILUS AND ITS ANTICANCEROUS ACTIVITY AGAINST COLON CANCER CELL LINE

Aysha Afrin P. F., Rekha A.* and Soundhari C.

Abstract

To study the, extraction and purification of bacteriocin from Streptococcs thermophilus and its antibacterial activity against pathogenic bacteria and anticancerous activity against colon cancer cell line. Bacteriocin was produced from S.thermophilus and it was purified by ammonium sulphate precipitation and various chromatographic techniques. The molecular weight was determined by Sodium dodecyl sulphate polyacrylamide gel electrophoresis. The cytotoxicity was checked using Vero cell lines and the anticancerous activity was checked using HT-29 colon cancer cell line. The isolate was identified as S.thermophilus based on morphological characteristics such as gram positive, non-motile, facultative anaerobe, catalase and oxidase negative and α-haemolytic bacteria. The bacteriocin was produced, purified and the protein was estimated by lowry’s method. The molecular weight of bacteriocin was determined as 20 K Da. The antibacterial activity was shown against Escherichia coli, Proteus, Salmonella and Shigella using agar well diffusion method. Cytotoxic effects was done on Vero cell lines and toxic free concentration was estimated as 250 μg. Anticancerous activity of bacteriocin was performed against HT-29 colon cancer cell line by MTT assay.

Keywords: Streptococcus thermophilus, Bacteriocin, Lowry’s, PAGE, Cytotoxicity, MTT.