RESUMO
Extracellular lipase from Beauveria bassiana strain CG481 was immobilized by using thirteen different immobilization protocols. Silica gel was chosen as the most suitable adsorbent with 94.8% of activity yield. The adsorption on silica gel did not change the optimum pH (8.5) and temperature (45ºC) values of the free lipase (FL) for lipolytic activity, and it showed higher activities in extreme conditions (pH 9.0 to 10.5, 60ºC). The lipase immobilized on silica gel (ILS) showed enhanced stability at pH 7.0 after 120 h incubation (69.0%) when compared to FL (33.3%). The thermal stability was also enhanced by immobilization at 60ºC in aqueous (64.6%) and organic medium (95.1%), while FL showed only 40.6% of residual activity in aqueous medium and exhibited no activity for esterification reaction in n-heptane. The treatment of ILS with 0.8 M NaCl prevented lipase desorption while Triton X-100 (0.1%) resulted the enzyme leakage. The ILS was reused for four times for esterification reaction with 80.8% of initial activity.
RESUMO
The objective of this study was to investigate the immobilisation efficiency of soybean β-glucosidase (181.6 U/mL; 23.8 mg protein/mL) on activated chitosan beads. Central Composite Rotational Design (CCDR) 23 was used and the application of immobilised enzyme in commercial soy drink was evaluated. The activation of chitosan beads was achieved with established 2.5% glutaraldehyde, pH 7.5, 8 h incubation time (6 h with agitation and 2 h without agitation) at 37ºC. The highest immobilisation efficiency (%) of soybean β-glucosidase on chitosan beads obtained was 37.74 U/mL and 18.84 mg protein/4 chitosan beads at pH 7.5 and 20 h coupling time of enzyme-matrix (7 h with agitation and 13 h without agitation) at 4ºC. The immobilised enzyme incubated at 50ºC, pH 5.5 resulted in 24% increase in the aglycones content in commercial soy drink after 60 min.