Site directed mutagenesis of alkyl hydroperoxide reductase.

Abstract

This research was designed to investigate the binding site of nicotinamide adenine dinucleotide (NADH) on alkyl hydroperoxide reductase subunit F (AhpF) of Salmonella typhimurium by using site directed mutagenesis. The postulated binding site was suggested by using molecular mechanics Poisson–Boltzmann surface area (MM/PBSA) approach.

AhpF enzyme requires NADH as a cofactor to function properly in peroxide reductase catalysis. The wild type AhpF was mutated in such a way that it could no longer bind to NADH as postulated by MM/PBSA approach. The hypothetical binding site would be proven correct if the activity of the mutant enzyme decreases significantly from the wild type.