Simultaneous purification and immobilization of soybean hull peroxidase with a dye attached to chitosan mini-spheres
- Bracco, Lautaro Fidel
- Levin, Gustavo Javier
- Navarro, Agustin Andres
- Wolman, Federico Javier
- Miranda, Maria Victoria
- Cascone, Osvaldo
DOIAbstract
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...
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