Process Biochemistry, Vol.48, No.10, 1516-1523, 2013
Cassava (Manihot esculenta Crantz of cv. KU50) peroxidase and its potential for the detection of some thiol compounds based on the inhibitory effect of 3,3',5,5'-tetramethylbenzidine oxidation
Cassava peroxidase (CSP) isolated from cassava leaves (Manihot esculenta Crantz of cv. KU50) was purified by DEAE and concanavalin-A column chromatography. CSP was a haem-containing cationic glycoprotein with the molecular weight of 38 and 44 kDa determined by MALDI-TOF-MS. Its kinetic catalysis in the presence and absence of some thiols were investigated and compared to those of horseradish and soybean peroxidases by using urea hydrogen peroxide (UHP) and 3,3',5,5'-tetramethylbenzidine (TMB) as substrates. Inhibitory effects of some pesticides containing thiol groups such as thiosemicabazide, thiourea and thiophanate-methyl on TMB oxidation were graphically demonstrated their ability to be either competitive or noncompetitive inhibitors, depending on their properties. The degrees of inhibition were expressed as K-i and IC50 values. The applicable range for the detection of thiosemicabazide in solution was found to be in the range of 10-100 mu M, whereas those of thiourea and thiophanate-methyl were 40-400 mu M and 10-460 mu M, respectively. On the contrary, the inhibitory effects of some phenolic pollutants; 4-nitrophenol, 4-phenylphenol and pentachlorophenol on TMB oxidation were not significantly observed and the phenomenon was discussed. (C) 2013 Elsevier Ltd. All rights reserved.