Comparative Analysis of the Kinetic Properties of Peroxidases from two Improved Sorghum Varieties
doi.org/10.26538/tjnpr/v5i7.21
Keywords:
Cereal enzymes, Enzymes and catalysis, Food and brewing chemistry, Protein purification, Sorghum and cereals, Sorghum peroxidaseAbstract
Varietal differences and their influence are not often critically considered when assessing properties of industrially important enzymes such as peroxidases, from cereals. Therefore, this study aims at closing that knowledge gap, by purifying peroxidases from two improved sorghum varieties, KSV8 and SK5912, which are food and brewery-grade grains used extensively all over the world. This was done via (NH4)2SO4 precipitation, ion exchange chromatography and gel filtration, and comparing their properties. Both KSV8 and SK5912 peroxidases had their soret peaks at 402 nm but with different extinction coefficients (ε 402 = 113 mm-1 cm -1 for KSV8 and ε402 = 119 mm-1 cm -1 for SK5912; while their molecular weights using ESI mass spectral data were 35.591 and 35.645 kDa, respectively. Optimum activity pH for KSV8 were pH 5 (guaiacol and o-dianisidine) and pH 4 for ABTS, pyrogallol pH 8, while for SK5912, they were pH 5 (guaiacol) and pH 4 (o-dianisidine, ABTS and pyrogallol). Optimum temperatures when measured with guaiacol were 50oC with KSV8 and 40oC for SK5912. Steady state kinetics of the enzymes with the same organic compounds showed that they have different affinities for them with o-dianisidine being the most efficiently oxidized substrate. Calcium greatly increased the activities of both sorghum peroxidases; however, the rate was much higher in SK5912 than in KSV8, while with ferric ion, the opposite obtained. Such differences or similarities were replicated across several other properties studied. These findings underline the great importance of differences in cereal varieties which should be reflected in their different biotechnological applications.
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