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

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. SDSPAGE 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.

Serratiopeptidase: An integrated View of Multifaceted Therapeutic Enzyme

“…Very few studies have been reported on the anti-cancer activity of serratiopeptidase. The in vitro cytotoxic activity of serratiopeptidase against colon cancer cell lines (Caco-2) was reported by Araghi et al [ 63 ]. The findings of previous reports suggested that the enzyme has anti-cancer potential, but further in vitro and in vivo mechanistic pathway studies are needed in order to confirm the biological activity of the enzyme.…”

Genomic analysis and plant growth-promoting potential of a Serratia marcescens isolated from food

“…Additionally, even new strategies proposed to treat antibiotic resistance, like bacterial predation, effective against some Gram-negative bacteria (Rames, 2020), find a barrier in our strain as a result of the metalloprotease serralysin that can reduce the ability of some predators to attach to the S. marcescens, conferring protection (Garcia et al, 2018). Serralysin, a virulence factor used medicinally as a proteolytic enzyme, is therapeutically useful in the management of pain and inflammation as a broad spectrum anti-inflammatory drug (Tiwari, 2017), digests non-living tissue, blood clots, cysts, and arterial plaque (Rouhani et al, 2020) and has an anticancer potential (Araghi et al, 2019).…”

Bioprospecting of serratiopeptidase-producing bacteria from different sources

“…The isolated strains were examined for protease activity on skim milk agar plates (10% skim milk powder, 0.5% peptone, 0.5% NaCl, and 1% agar) (Jo et al, 2008). The primary screening was performed with 10% skim milk powder (Araghi et al, 2019). After a 24 h incubation at 37°C, the plates were checked for the zone of hydrolysis (Masi et al, 2021).…”