Purification and Characterization of α-Amylase from Bacillus cereus SM22 Isolated from Deteriorating Cocoyam doi.org/10.26538/tjnpr/v6i2.9
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Abstract
Amylases can be sourced through various organisms that thrive on common food items such as cocoyam. This study optimized, purified, and characterized α-amylase from Bacillus cereus SM22 isolated from decayed cocoyam tubers. Healthy cocoyam tubers were homogenized, filtered, allowed to stand, and then decanted. The residue was dried at 50oC for 48 hours, powdered and used as cocoyam starch. Inoculum size, temperature, pH, starch concentration, nitrogen and carbon sources were used to optimize the α-amylase activity/production by Bacillus cereus SM22. The α-amylase activity and protein concentration of the cell-free culture supernatant were determined. The supernatant was purified on DEAE-Sephacel and BioGel P-100 and the native molecular weight of the α-amylase was estimated. Also, the α-amylase was optimized and kinetic parameters; Michaelis constant (Km) and maximum velocity of the reaction (Vmax) were determined. The result showed that pH 7.0, 37oC, 1% cocoyam starch, 2% inoculum, peptone and ammonium nitrate were necessary for α-amylase production by Bacillus cereus SM22. Similarly, pH 6.0, 50oC, NaCl (100 or 240 mM), Mg2+ (80-120 mM) and Ca2+ (20-80 mM) were required for maximum α-amylase activity. However, Al3+ and EDTA completely inhibited the α-amylase activity. The study successfully purified and optimized α- amylase from Bacillus cereus SM22 isolated from decayed cocoyam tubers. It demonstrated the necessary factors for optimal performances of Bacillus cereus SM22 and α-amylase in a given culture medium. Also, the low Km and high Vmax for starch substrate make the α-amylase from Bacillus cereus SM22 an excellent, cheap, and robust candidate for industrial applications.
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