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

BRACCO, LAUTARO F.; LEVIN, GUSTAVO J.; NAVARRO DEL CANIZO, AGUSTIN A.; WOLMAN, FEDERICO J.; MIRANDA, MARÍA V.; CASCONE, OSVALDO

BIOCATALYSIS AND BIOTRANSFORMATION

TAYLOR & FRANCIS LTD

Lugar: Londres; Año: 2017 vol. 5 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 behavior 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 remainingactivity 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 % of2-bromophenolfrom wastewaterin 180 min and, after 5 cycles of use, the activity loss was only 12.8 %.