Simultaneous purification and immobilization of soybean hull peroxidase with a dye attached to chitosan mini-spheres |
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| Authors: | Lautaro Fidel Bracco Gustavo Javier Levin Agustín Andrés Navarro del Cañizo Federico Javier Wolman María Victoria Miranda |
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| Institution: | 1. Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina;2. NANOBIOTEC, Consejo Nacional de Investigaciones Científicas y Técnicas (UBA-CONICET), Buenos Aires, Argentina;3. Centro de Investigación y Transferencia de Entre Ríos (CITER) CONICET- UNER, Gualeguaychú, Entre Ríos, Argentina |
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| Abstract: | Soybean hull peroxidase (EC 1.11.1.7, SBP) was simultaneously purified and immobilized by dye affinity chromatography with Reactive Blue 4 attached to chitosan mini-spheres. Under optimized conditions, 96% of SBP was adsorbed to the matrix. Under the most stringent condition, only 49% was desorbed, whereas 2 M NaCl failed to desorb a significant amount of SBP. This behaviour allowed proposing the dye matrix as a support to immobilize SBP from a crude extract. The pH of maximum activity shifted from 7 to 3–5. SBP gained thermostability after immobilization: after 5?h at 85?°C, the remaining activity was 54%, whereas that of the free enzyme was 31%. The optimum temperature for the immobilized SBP was 75?°C, whereas that of the free enzyme was 55?°C. After two months at 4?°C, the activity loss of the immobilized SBP was only 3%. Immobilized SBP removed 80% of 2-bromophenol from wastewater in 180?min and, after five cycles of use, the activity loss was only 12.8%. |
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| Keywords: | Soybean peroxidase purification immobilization affinity chromatography triazine dyes bioremediation |
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