Development of a heavy metals enzymatic-based assay using papain

A heavy metals enzymatic-based assay using papain was developed. Papain was assayed using the Casein-coomassie-dye-binding assay. The assay is sensitive to several heavy metals. The IC50 (concentration of toxicant giving 50% inhibition) of Hg2+, Ag2+, Pb2, Zn2+ is 0.39, 0.40, 2.16, 2.11 mg l−1, respectively. For Cu2+ and Cd2+ the LOQ (limits of quantitation) is 0.004 and 0.1 mg l−1, respectively. The IC50 and LOQ values were found to be generally comparable to several other enzymatic and bioassays tests such as: immobilized urease, 15-min Microtox™, 48 h Daphnia magna, and 96 h Rainbow trout. The papain assay is xenobiotics tolerant, has a wide pH for optimum activity, is temperature stable, and has a relatively quick assay time. The papain assay was used to identify polluted water samples from industrial sources in Penang, Malaysia. We found one site where the assay gave a positive toxic response. The toxicity of the site was confirmed using Atomic Emission Spectrometry analysis.