In the present study, a mixed culture from a local agricultural soil sample was isolated for Metanil Yellow (MY) dye decolorization. The metagenomic analysis confirmed that 42.6% has been dominated by genus Bacillus, while Acinetobacter (14.0%) is present in the microbial communities of the mixed culture. For fungi diversity analysis, around 97.0% was “unclassified” fungi and 3% was Candida. The preliminary investigation in minimal salt media (MSM) showed that 100% decolorization was achieved after 24 h of incubation. Response surface methodology (RSM) was successfully applied using Box-Behnken design (BBD) to study the effect of four independent parameters—MY dye concentration, glucose concentration, ammonium sulfate concentration, and pH—on MY dye decolorization by the mixed bacterial culture. The optimal conditions predicted by the desirability function were 73 mg/L of MY, 1.934% glucose, 0.433 g/L of ammonium sulfate, and a pH of 7.097, with 97.551% decolorization The correlation coefficients (R2 and R2 adj) of 0.913 and 0.825 indicate that the established model is suitable to predict the effectiveness of dye decolorization under the investigated condition. The MY decolorization of the mixed bacterial culture was not affected by the addition of heavy metals in the growth media. Among the 10 heavy metals tested, only copper gave 56.19% MY decolorization, whereas the others gave almost 100% decolorization. The decolorization potential of the mixed bacterial culture indicates that it could be effective for future bioremediation of soil-contaminated sites and treatment solutions of water bodies polluted with the MY dye.
In this study, the Metanil Yellow (MY) decolorizing mixed culture, namely FN3, has been isolated from agriculture soil. The mixed culture was immobilized using gellan gum. In order to optimize the immobilization process for maximal dye decolorization, Response Surface Methodology (RSM) was performed. The optimal conditions for immobilization predicted by desirability function are 130 mg/L of MY dye concentration, 1.478% of gellan gum concentration, 50 beads and 0.6 cm of beads size with the percentage of decolorization of 90.378%. The correlation coefficients of the model (R2 and R2 adj) are 0.9767 and 0.9533, respectively. This indicates that the established model is suitable to predict the effectiveness of dye decolorization under the investigated condition. The immobilized beads of mixed culture FN3 were able to be reused up to 15 batches of decolorization. The immobilized cells also have high tolerance towards heavy metals. This was proven by higher dye decolorization rate by the immobilized cells even with the addition of heavy metals in the media. The decolorization potential of the mixed culture indicates that it could be useful for future bioremediation of soil contaminated sites and treatment solutions of water bodies polluted with MY dye.
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