Optimization of concentration parameters for maximizing bacteriophage recovery value using response surface methodology: A case study on determining the optimal concentration parameters for Erwinia amylovoraEA1T1.B3 phage

Abstract

Bacteriophages have emerged as effective antimicrobial agents in combating pathogenic bacteria. To successfully apply phages in real‐life scenarios, it is crucial to optimize their large‐scale production, concentration and purification processes. In this study, our objective was to optimize the phage concentration parameters with a high recovery rate within a shorter time and minimum chemical consumption. We isolated and characterized a specific Erwinia amylovora phage. Subsequently, we employed response surface methodology (RSM) combined with a three‐factor central composite design to optimize the phage recovery. The factors considered were polyethylene glycol (PEG) concentration, NaCl concentration and incubation time. The optimized values for PEG, NaCl and incubation time, which resulted in a maximum recovery rate of 85.37%, were determined as 18%, 2.38 M and 0 h, respectively. This finding indicates that the concentration step, which traditionally took up to 18 h with a recovery rate of 65%, can now be accomplished in a significantly shorter time with an improved recovery rate. Our study demonstrates the effectiveness of RSM to identify the optimum conditions for phage precipitation. This approach allows for the rapid and efficient design of precipitation protocols tailored to specific bacteriophages, resulting in shorter processing times and higher recovery rates.