Extraction and Purification of Anticancer Enzyme: Protease from Pseudomonas aeruginosa

Abstract

This study investigated the anticancer and wound-healing potential of protease enzymes derived from Pseudomonas aeruginosa against cancer and normal cell lines. Out of 110 bacterial isolates obtained from clinical samples like urine, burns, and wounds, 67 were identified as Pseudomonas aeruginosa using biochemical tests and the VITEK-2 Compact system. Qualitative and quantitative screening revealed that 56 isolates produced protease. Notably, clinical isolate 9A from burn and wound samples demonstrated high protease activity, with specific activities of 280U/mg, indicating potential cytotoxic and wound healing properties. Optimal conditions for protease production were found to be at 37°C and pH 8 in Trypticase soy broth. The extracted enzymes underwent purification via gel filtration using Sephadex G-150 chromatography, resulting in significant purification folds for the protease. Cytotoxicity assays using MTT revealed that the protease showed increased cell viability, indicating possible growth-promoting effects. However, excessive protease activity led to cytotoxicity in normal cell lines, highlighting the need for controlled enzyme activity. Moreover, in vitro scratch tests demonstrated that protease showed a further pronounced decrease in wound area, revealing its potential role in processes of wound-healing.