Use of Aqueous Two-Phase System for Partial Purification and Characterization of α-Amylase from Burnatia enneandra Micheli

Alpha-amylase from Burnatia enneandra Micheli, a tuber plant from a rural area of Cameroon, extracted under optimal conditions was purified using aqueous two-phase extraction system (ATPS). The separation behaviour was further investigated. Polyethylene glycol (PEG, % w/w), ammonium sulphate (AS, % w/w) and pH were optimized for the purification of B. enneandra alpha-amylase. Optimum conditions for purification factor were PEG, 13.1 %, AS, 12 % and pH 4. Highest enzyme recovery was achieved at PEG, 13.0 %, AS, 12 % and pH 5.1. Further, enzyme characterization surveys acknowledged that alpha-amylase formed was just an acidic enzyme. The pH and temperature at which the enzyme was well expressed were 5.5 and 70 °C respectively. Another characteristic of the enzyme was determined like the Michaelis-Menten constant (Km) and Vmax which value obtained was respectively 89.32 μg/mL and 2.99 μg/mL/min. Practical application The alpha-amylase obtained from local Burnatia enneandra Micheli tuber can be used for brewing purpose and other field using starch (fruit juice, spirit, sirop etc). This is quite important as none of the amylase enzymes used by factories in Cameroon is from local source.