Three-phase partitioning for isolating peroxidase from lemon peels

Abstract

Background and objective: Peroxidase is an oxidoreductase that uses different compounds as substrates and thus can be utilized for different applications. The goal of this work is to isolate peroxidase from lemon peels using Three-Phase Portioning (TPP). Methods: TPP was set by adding varying amounts of salts and alcohol and the enzyme activity recovery was measured for each variable. Different parameters were optimized successively in order to achieve the highest enzyme activity recovery including salt type, salt concentration, pH, alcohol/crude extract ratio and type of alcohol and then, combining all optimized conditions together. Results: Salt that gave maximal recovery was sodium potassium tartrate, optimal salt concentration was 15%, optimal pH was 8, optimal alcohol/crude extract ratio was 1 and t-butanol was preferred to 1-butanol. Efficiently, upon combining all optimized factors, an activity recovery of 175% was obtained. Conclusion: This protocol provides an easy, feasible method to efficiently isolate peroxidase from lemon peels using TPP.