In Vitro and in Silico Analysis of Recombinant Arginine Deiminase from Pseudomonas Furukawaii as a Potential Anticancer Enzyme

Abstract

Arginine deiminase (ADI) is a promising anti-tumor enzyme that can be employed in amino acid deprivation therapy for the treatment of various arginine auxotrophic tumors. In our previous work, Pseudomonas furukawaii was identified as a potent producer of ADI with optimum activity at physiological pH and temperature. In this study ADI coding arcA gene was cloned and expressed in E. coli BL21. Recombinant ADI of P. furukawaii (PfADI) was purified using affinity chromatography and its molecular mass was estimated to be ~46KDa. PfADI was found to effectively inhibit the HepG2 cells with an IC50 value of 0.1950 IU/ml which is lower than the IC50 value of Mycoplasma hominis ADI (MhADI) which is in phase III clinical trials. The 3D structure of PfADI was modeled. Immunoinformatics analysis was also carried out to compare the immunogenicity of PfADI with MhADI. The PfADI was found to be less immunogenic in terms of number of linear and conformational B cell epitopes and T cell epitope density. The overall antigenicity and allergenicity of PfADI was also lower as compared to MhADI. Thus the present in vitro and in silico studies establish PfADI as a potential anticancer drug candidate with improved efficacy and low immunogenicity.