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Purification and Characterization of a Protease Inhibitor with Anticancer Potential from Bacillus endophyticus JUPR15

[ Vol. 15 , Issue. 1 ]

Author(s):

Prerana Venkatachalam* and Varalakshmi Kilingar Nadumane   Pages 74 - 82 ( 9 )

Abstract:


Introduction: Protease Inhibitors (PIs) constitute a group of proteins widely distributed among all organisms and their main function includes their ability to inhibit the proteolytic activity. PIs represent an important role in the regulation of various cellular physiological and biological processes, including cell cycle, cell death, differentiation and immune response.

Material and Methods: Hence, in our search for novel anticancer compounds, we isolated microorganisms from various environmental sources and screened them for the production of protease inhibitors. Promising isolates were further checked for their protease inhibitory activity by their ability to inhibit the activity of trypsin and chymotrypsin, which were measured spectrophotometrically.

Results: The isolate identified as Bacillus endophyticus JUPR15 was found to be promising with higher inhibitory activity than the other isolates. The inhibitor was purified by cold acetone precipitation and column chromatography and further subjected to characterization studies by performing 12 % SDS-PAGE to determine the molecular weight and gelatin-PAGE assay to confirm its inhibitory activity.

Conclusion: The isolate exhibited promising anticancer activity on in-vitro Hela and HepG2 cancer cell lines, showing its application potentials.

Keywords:

Bacillus endophyticus JUPR15, anticancer, protease inhibitors, Gelatin-PAGE, MTT assay, enzyme kinetics.

Affiliation:

Department of Biotechnology, School of Sciences, Jain University, Jayanagar, Bengaluru-560 011, Department of Biotechnology, School of Sciences, Jain University, Jayanagar, Bengaluru-560 011

Graphical Abstract:



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