Characterization of Partial purified alpha-amylase enzyme produced from locally isolate Bacillus licheniformis
journal of kerbala university,
2017, Volume 13, Issue 1, Pages 126-138
AbstractAlpha-amylase enzyme produced from No.5 isolate which its efficiency in production among 24 isolates as Bacillus spp .This isolates was identified depending on the morphological character ,microscopical and biochemical tests . The crud enzyme extract which produced from isolated in this study wwere partial purified starting by using 60% saturation ammonium sulphate precipitation and dialysis , followed by batch manner ion exchange by using DEAE-Sepharose CL-6B and finally gel filtration step by using Sephadex G-75 The yield enzyme was 34.144% and the purification fold was 4.286. The molecular weight of enzyme was estimated to 56.25 KD as then gel filtration nose and way above. The optimum pH of enzyme activity was 7, whil the rang of pH stability from 5 to 8. The optimum temperature for enzyme activity was 50 c̊ ,while the rang of heat stability was from from 20-60 c̊. Mechalis mention constant (Km) and maximum velocity (Vmax) valus were 7.143 mg/ml and 66.666 mg/ml/min respectively.The effect of Calcium and magnesium chloride metal ions were activator for enzymes produced. Maltose sugar was Relative flaw Rf value equal to 0.5 is the main product of hydrolysis of starch by alpha-amylase enzyme that’s partial purified by using thin layer Chromatography (TLC) Thin Layer Cromotography technique.
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