Simultaneous purification and immobilization of soybean hull peroxidase with a dye attached to chitosan mini-spheres

LAUTARO FIDEL BRACCO; GUSTAVO JAVIER LEVÍN; AGUSTÍN A. NAVARRO DEL CAÑIZO; FEDERICO JAVIER WOLMAN; MARÍA VICTORIA MIRANDA; OSVALDO CASCONE

BIOCATALYSIS AND BIOTRANSFORMATION

TAYLOR & FRANCIS LTD

Lugar: Londres; Año: 2017 vol. 35 p. 306 - 314

1024-2422

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 5h at 85ºC, the remaining activity was 54%, whereas that of the free enzyme was 31%. The optimum temperature for the immobilized SBP was 75ºC, whereas that of the free enzyme was 55ºC. After two months at 4ºC, the activity loss of the immobilized SBP was only 3%. Immobilized SBP removed 80% of 2-bromophenol from wastewater in 180min and, after five cycles of use, the activity loss was only 12.8%.