Physicochemical Properties of Thiaminase from Cassava Tubers (Manihot esculenta)
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Abstract
Thiaminase (EC 2.5.1.2) is an antinutritional enzyme that is responsible for thiamine (vitamin B1) degradation, causing severe neurological and metabolic disorders in mammals. Cassava tubers were used for the extraction and purification of thiaminase using ammonium sulphate precipitation to 85 % and the CM-Sephadex C-25 chromatography. The purification process gave a specific activity of was 0.511 U/mg, 6.59 % yield and 7.61 fold-purification. The apparent Michaelis-Menten constant (Km) values were 0.20 μM for thiamine and 0.23 μM for the co-substrate aniline, showing a much greater affinity for thiamine. The maximum velocities for both substrates were 0.0556 ± 0.01 μmol/min/mL and 0.107 ± 0.05 μmol/min/mL respectively. The optimum temperature and pH of the enzyme were 50 °C and 5.0 respectively. The enzyme activity was activated by low concentrations (0.1 mM and 1.0 mM) of Fe2+, Hg2+, Mn2+ and Na+ and also, a remarkable enhancement of the enzyme activity was exhibited with 10 mM concentration of all these cations indicating adaptation to the metallic profile of the soil. The enzyme was found to be stable at 40 °C to 60 °C. The findings of this study confirmed the presence of thiaminase in cassava tubers, suggesting that its activity may contribute to thiamine deficiency among populations that depend on cassava as a staple food, particularly when it is consumed raw or inadequately processed.
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