Fusion Proteins in the Fight against Colorectal Cancer: Assessing the Cytotoxic Potential of Recombinant Nisin-Arginine Deiminase Fusion Protein

Background: A promising strategy in cancer therapy involves the production of fusion proteins, which entail the fusion of two distinct proteins. This study aimed to produce and assess the cytotoxic effects of the Nisin-arginine deiminase (ADI) fusion protein on the SW480 cell lines, a common model for studying colorectal cancer (CRC). Materials & Methods: The designed Nisin-ADI gene fragment sequence was sent to Biomatik Company for synthesis in pET-28a vectors between SacI and HindIII restriction enzyme sites. Escherichia coli (E. coli) DH5α and BL21 were utilized for cloning and protein expression, respectively. The recombinant fusion protein expression was induced by Isopropyl ß-D-1-thiogalactopyranoside (IPTG) and purified using Ni2+-nitrilotriacetic acid (1) resin affinity chromatography. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were conducted to analyze the purified protein. The cytotoxic effect of the purified recombinant fusion protein on SW480 and NIH3T3 cells, as a control, was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Findings: The results of this study showed that the fusion protein had a significant impact on the SW480 cell lines. The Half-maximal inhibitory concentration (IC50) of the fusion protein was 30 µg/mL, indicating that it effectively inhibited the growth of cancer cells. However, the fusion protein did not significantly affect the control group. Conclusion: This study provides helpful insights into the potential application of recombinant Nisin-ADI fusion proteins as a potential treatment option for colorectal cancer. The potential for selective targeting of cancer cells is promising as normal cells are unaffected by this fusion protein.

Fusion Proteins in the Fight against Colorectal Cancer: Assessing the Cytotoxic Potential of Recombinant Nisin-Arginine Deiminase Fusion Protein Keywords:

Nisin , Arginine deiminase , Fusion protein , Colorectal cancer , Cytotoxicity