Improvement of Bacillus Amyloliquefaciens Strain for Enhancement of α- Amylase Production
1496 WordsJun 18, 20186 Pages
INTRODUCTION α-Amylases(E.C. 188.8.131.52.) are starch-degrading enzymes that catalyze the hydrolysis of internal a-1,4-O- -glycosidic bonds in polysaccharides with the retention of a-anomeric configuration in the products. Bacillus amyloliquefaciens find potential application in a number of industrial processes such as in food, fermentation, textiles and paper industries . Most of the a-amylases are metalloenzymes, which require calcium ions (Ca2+) for their activity, structural integrity and stability. The enzymatic hydrolysis is preferred to acid hydrolysis in starch processing industry due to a number of advantages such as specificity of the reaction, stability of the generated products, lower energy requirements and elimination of…show more content…
at 150 rpm in shaker incubator at 370 C.
Different pH Stresses on Metal Stressed Culture With Maximum α-amylase Activity:Effect of different pH (5.5, 6, 6.5, 7, 7.5, 8.0,) on alpha amylase production by the strain of Bacillus amyloliquefacienswas investigated. The hydrolysis of starch by α-amylase is highly affected by pH.In present investigation, the different pH (5.5-8.0) was tested for the activity of α-amylase.
Different Salt Stresses on pH Stressed Culture With Maximum α-amylase Activity: The salt at different amount was given to Bacillus amyloliquefaciens to act as supplement for amylase production. In present study different amount like 1%, 1.5%, 2% and 2.5% were used and produced amylase was studied followed by DNS enzyme assay method.
EXTRACTION OF ENZYME: After optimum incubation period the experimental test-tube was harvested. The fermented biomass sample was centrifuged at 10,000 rpm for 15minutes in the centrifuge to remove the spores of the organism. The supernatant was carefully collected and the crude enzyme, thus obtained, was subjected to enzyme assay.
ESTIMATION OF ENZYME ACTIVITY BY DNS METHOD By constructing a standard curve graph for maltose and estimated the concentration of Maltose present in an unknown sample used for determining the activity of amylase enzyme. Maltose reduces the pale yellow colored alkaline 3, 5-Dinitro