Immobilization of cauliflower myrosinase on agar agar matrix and its application with various effectors

The purified and thermo stable myrosinase from cauliflower seedlings was immobilized by 3.5% agar agar matrix and immobilized myrosinase chips were stored in extraction buffer. After 48 hours 30% enzyme activity was exuded into extraction media from immobilized enzyme chips. Effect of some metal ions and organic solvents on the activity of immobilized cauliflower myrosinase was studied. Amongst selective metal ions (Sr 2+ , Sn 2+ and Ba 2+ ), Sr 2+ at 4 mM concentration exhibited marked activating effect on the activity up to 3 fold. However, Sn 2+ and Ba 2+ increased the activity to a certain extent and then suppressed. Activation kinetics of myrosinase in presence of Sn 2+ and Sr 2+ were studied between 0-20min.The rate of reaction was almost constant till 15 min and then slight deactivation was recorded at various concentrations used. On the other hand, few heavy metal ions [Fe 2+ , Fe 3+ , Cu 2+ and Zn 2+ ] strongly inhibited the activity even at lower concentrations. Several nonpolar organic solvents even at comparatively higher concentrations had detectable activation effects. Further, their activity was seen with respect totime (27 min). However, some protic polar organic solvents exhibited inhibitory effect with immobilized myrosinase except to butanol.