Enzymatic detection of heavy metal ions in aqueous solution from vegetable wastes by immobilizing pumpkin (Cucumis melo) urease in calcium alginate beads

Enzyme urease is extracted from the discarded seeds of pumpkin. Urease was purified to apparent homogeneity (5.2 fold) by heat treatment at 48 +/- 1 degrees C and gel filtration through Sephadex G-200. Effect of model metal ions on the activity of the homogeneous enzyme preparation (sp. activity 353 U/mg protein, A(280)/A(260) = 1.12) of soluble as well as immobilized enzyme was investigated. The soluble and immobilized urease has been used for the quantitative estimation of general water pollution with heavy metal ions like Hg2+, Cu2+, Cd2+, and Co2+. The measurements of the urease residual activity have been carried out in tris-acetate buffer after pre-incubation of model metal salt. The inhibition was found to be biphasic with an initial rapid loss of activity and remainder in slow phase of 10 similar to 15 min. The immobilization was done in 3.5% alginate beads leading to 86% of entrapment. There was no leaching of the enzyme over a period of 15 days at 4 degrees C. The beads were fairly stable up to 50 degrees C and exhibited activity even at -10 degrees C. The inhibition by these ions was non-competitive and irreversible, hence could not be restored by dialysis. Based on the values of inhibition constant K; the heavy-metal ions were found to inhibit urease in the following order Hg2+ > Cu2+ > Cd2+ > Co2+. (C) KSBB.