Bacteriocin-producing strains were isolated from the soil of the rice field, screened out using an agar-well diffusion assay against six indicator bacterial strains, and the highest among them was selected for further investigation. The study focuses on how different growing conditions affect bacteriocin production. One-parameter-at-a-time (OPAT) and a central composite design of response surface methodology (RSM) were used to perform the optimization in two steps. In the OPAT trials, bacteriocin synthesis was elevated by 29%, 45%, and 34%, by employing sucrose as a carbon source and changing the NaCl concentration and pH at 7, respectively. To determine the linear, squared, and interaction correlations among the process variables to predict the ideal conditions for production, a four-factor central composite design (CCD) of RSM was used. It was determined that the analysis of variance (ANOVA), which produces a recognized model using RSM, is sufficient to describe bacteriocin production regarding activity (R2 = 0.9606). The ideal conditions for increased production were 1.51% sucrose concentration, 1.59% NaCl concentration, pH 6.35, and 28.66 (about 29) hours of incubation. The value predicted by RSM (4051.55 AU/mL) was approximately two times greater than the value of a non-optimized medium. The experimental value of 4403.85 AU/mL was closer to the expected value. According to the data, increasing bacteriocin activity required employing the ideal sucrose concentration, NaCl concentration, and incubation time. The partially purified bacteriocin was found stable at temperatures between 24 and 50 °C and at pH 5-8. The molecular weight purified bacteriocin was determined to be between 13 and 35 kDa.
Keywords: Bacteriocin; Central composite design; One-parameter-at-a-time; Optimization; Response surface methodology.
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