Application of central composite design approach for optimization nitrate removal from aqueous solution by immobilized Pseudomonas putida.

Abstract

High nitrate concentration is a dangerous pollutant in the environment. Immobilization for the optimum denitrifying bacterial strain isolated from collected wastewater samples was suggested for bioremediation of excessive nitrate concentration from aqueous solutions and its denitrification activity under different pH, nitrate concentration, bacterial beads, temperature and sodium alginate concentration conditions was explored. The active isolate was identified as Pseudomonas putida MT364822.1 by 16S rRNA analysis. Nitrate bioremediation process was optimized by applying response surface methodology based on central composite design approach. Nitrate uptake was significantly affected by variables of study (P-value <0.05). Maximum removal of nitrate (91.1%) was achieved at pH 7, nitrate concentration 400 mg/L, immobilized bacterial beads 3.0 g/L, temperature 35 °C and sodium alginate concentration 2.5% as optimal variable values. For application, immobilized P. putida MT364822.1 removed 82.2% of nitrate from raw fish farm effluent. Storage and reusability experiments showed that the immobilized strain stronger and more stable than the pure strain. The results suggested that immobilized P. putida MT364822.1 is a highly promising and suitable microorganism for use in the bio-removal of nitrate, and the central composite design was more effective in optimizing variables to achieve the best nitrate removal efficiency.