Journal of Biotechnology & Biomaterials
Open Access

Abstract

Chemotherapy is one of the conventional treatment methods employed for cancer therapy. The various challenges faced by chemotherapy are side effects of the drugs due to non-selectivity and high toxicity. Hence, it becomes imperative to seek alternative approaches focusing on targeted drug delivery to tumor cells. Attempts to increase the selectivity of drug are currently an intensively studied research aspect. Designing and developing prodrug is a strategy to minimize toxicity based on improvement of drug selectivity. A recent promising approach of targeted drug delivery of the anticancer agent is gene directed enzyme prodrug therapy (GDEPT) which involves prodrug activating enzymes. Nitroreductases are the enzymes employed in GDEPT which reduces aromatic nitro groups to hydroxylamines which are potent cytotoxins. The present research aims to characterize and engineer Pseudomonas aeruginosa nitroreductase enzyme for higher catalytic activity towards prodrug CB1954 (dinitroaziridinylbenzamide). Pseudomonas aeruginosa PAO1 nitroreductase (Pseudo_NR) consists of 200 amino acids. The present study envisages sequence analysis, taxonomic distribution and comparison of Pseudo_NR with structural homologues (FMN binding sites, dimer interface residues), homology modeling and docking studies with prodrug CB1954. It was observed that the FMN binding site in case of Pseudo_NR is more similar to that of B. subtilis, T. thermophilus and S. pneumoniae nitroreductase. However the dimer interface sites are observed to be variable in nature. The 3D simulation model of Pseudo_NR indicates that it is homo dimer and closely related to Bacillus subtilis nitroreductase ydfN and can be fitted into the X-ray structure with root mean square deviation of 0.61 Ã?Â?. The binding sites identified by docking studies will be investigated further for improved activation with prodrug CB1954.

Biography

Email: razkutty@yahoo.co.in