Purification and study of anti-cancer effects of Serratia marcescens serralysin

  • Almas Araghi Department of Microbiology, Faculty of Biological Sciences, North Tehran Islamic Azad University, Tehran, Iran
  • Saba Hashemi Department of Pharmaceutical Biotechnology, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  • Abbas Akhavan Sepahi Department of Microbiology, Faculty of Biological Sciences, North Tehran Islamic Azad University, Tehran, Iran
  • Mohammad Ali Faramarzi Department of Pharmaceutical Biotechnology, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
  • Mohsen Amin Department of Drug and Food Control, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran AND The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences, Tehran, Iran
Keywords: Serratia marcescens, Serralysin, Purification, Ion exchange chromatography, Anti-cancer

Abstract

Background and Objectives: Serralysin is an extracellular metalloprotease from Serratia marcescens which has been the subject of extensive biological investigations. The goal of this study was to extract and purify serralysin from S. marcescens and to investigate its cytotoxic activity on the colorectal cancer cell line.
Materials and Methods: The presence of the serralysin gene was confirmed using PCR. The supernatant of bacterial culture was collected and precipitated using ammonium sulfate. The precipitated protein was dialyzed and subjected to ion exchange chromatography for further purification. Casein assay and skim milk assay was used to confirm the enzymatic activity. SDS-PAGE was used to visualize the presence of serralysin. Metalloprotease inhibition activity was performed using 50 mM EDTA. Cytotoxic activity of serralysin was assessed on MTT assay.
Results: The PCR product corresponding to serralysin was estimated to be approximately 1500 bp. A transparent zone around the bacterial colonies on skim milk agar and casein digestion confirmed the proteolytic activity of serralysin. A 52 kDa band in SDS-PAGE corresponding to serralysin was observed before and after purification processes. MTT assay showed IC50 values 24.78 μg/ml and 19.16 μg/ml after 24 h and 48 h exposure of Caco-2 cells to serralysin, respectively.
Conclusion: Our results showed that native serralysin has anticancer potential and may be a candidate for further pharmaceutical research and development. Further in vivo and in vitro mechanistic studies are suggested to confirm the biological activities.

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Published
2019-09-17
How to Cite
1.
Araghi A, Hashemi S, Akhavan Sepahi A, Faramarzi MA, Amin M. Purification and study of anti-cancer effects of Serratia marcescens serralysin. Iran J Microbiol. 11(4):320-327.
Section
Original Article(s)