title: Kinetic characterization, thermo-stability and Reactive Red 195A dye detoxifying properties of manganese peroxidase-coupled gelatin hydrogel.
abstract: An indigenous and industrially important manganese peroxidase (MnP) was isolated from solid-state bio-processing of wheat bran by white-rot fungal strain Ganoderma lucidum IBL-05 under pre-optimized growth conditions. Crude MnP extract was partially purified (2.34-fold) to apparent homogeneity by ammonium sulphate precipitation and dialysis. The homogeneous enzyme preparation was encapsulated on gelatin matrix using glutaraldehyde as a cross-linking agent. Optimal conditions for highest immobilization (82.5%) were: gelatin 20% (w/v), glutaraldehyde 0.25% (v/v) and 2 h activation time using 0.6 mg/mL of protein concentration. Gelatin-encapsulated MnP presented its maximum activity at pH 6.0 and 60 °C. Thermo-stability was considerably improved after immobilization. The optimally active MnP fraction was tested against MnSO4as a substrate to calculate kinetic parameters. More than 90% decolorization of Sandal-fix Red C4BLN (Reactive Red 195A) dye was achieved with immobilized MnP in 5 h. It also preserved more than 50% of its original activity after the sixth reusability cycle. The water quality parameters (pH, chemical oxygen demand, total organic carbon) and cytotoxicity (brine shrimp and Daphnia magna) studies revealed the non-toxic nature of the bio-treated dye sample. A lower Km, higher Vmax, greater acidic and thermal-resistant up to 60 °C were the improved catalytic features of immobilized MnP suggesting its suitability for a variety of biotechnological applications.
authors: Industrial Biotechnology Laboratory, Department of Biochemistry, University of Agriculture, Faisalabad, Pakistan E-mail: bilaluaf@hotmail.com; State Key Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Muhammad
authors: M
authors: Bilal
authors: Industrial Biotechnology Laboratory, Department of Biochemistry, University of Agriculture, Faisalabad, Pakistan E-mail: bilaluaf@hotmail.com.
authors: Muhammad
authors: M
authors: Asgher
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Hongbo
authors: H
authors: Hu
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Xuehong
authors: X
authors: Zhang
mesh_terms: D001391:Azo Compounds / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D005780:Gelatin / Q000737:chemistry; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007700:Kinetics; D009282:Naphthalenesulfonates / Q000737:chemistry*; D010544:Peroxidases / Q000737:chemistry*; D020940:Reishi / Q000737:chemistry; D014865:Waste Disposal, Fluid
chemical_list: D001391:Azo Compounds; D004396:Coloring Agents; D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D009282:Naphthalenesulfonates; C545303:reactive red 195; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D005780:Gelatin; D010544:Peroxidases; C051129:manganese peroxidase