Abstract
Thermostable amylase enzyme has a broad commercial value in its use in the processing of starch, sugar production, textile, paper, animal feed, pharmaceuticals and in the manufacture of detergents. This study aims to determine the optimum conditions of amylase production from the termofil bacteria Bacillus sp. RSII-1b isolated from a hot spring Lejja South Sulawesi and characterizing the amylase enzyme. The testing of amylase production was done with various concentration of starch and CaCl2 in the production medium, then fermented to obtain maximum amylase activity, amylase enzyme was produced in optimum condition, and its characteristic was tested using 2% starch substrates in various pH, temperature and determining the compound cofactor which can act as activators or inhibitors of the amylase activity, the enzyme activity was tested using DNS method. Crude extract enzyme has the highest enzyme activity of the protein content determined by the method of Lawry. The results showed that the amylase enzyme from Bacillus sp RSAII-1b isolates can be manufactured to a maximum at 33 hours of fermentation time with conditions: the concentration of substrate (starch) 1.5%, 0.08% CaCl2, 55°C temperature, medium pH 7.0 and aeration speed 200rpm with the activity of 0.1323U/mL, amylase crude extract protein content of 1.86mg/mL, with spesifik activity 0.0711 U/mg protein. Crude extract amylase work optimally at pH 6.0; 55°C -60°C the amylase activity of 0.165U/mL, the specific activity of 0.089U/mg protein. Amylase enzyme is an enzyme that depends on metal because its catalytic activity can be activated by metal ions Ca2+, Mg2+, Cu2+, Ni2+ and Co2+ as activators whereas Zn2+ ions decrease the activity of enzymes as inhibitors. Amylase activity in the crude extract optimum conditions with the addition of 10 mm ions Ca2+ can increase amylase activity up to 32,89%, while the addition of ions Zn2+ can inhibit amylase activity up to 25%.
Highlights
Starch molecules are polymers of alpha-D-glucopyranose which will be broken down by amylase enzyme at alpha-1,4 and alpha-l,6-glycosides bonds [1]
Amylase is a hydrolytic enzyme that can hydrolyze the compound starch or starch as its substrate, and an enzyme that is important and its existence is greatest in the field of food and biotechnology, these enzymes are traded as much as 25% of the total other enzyme [2]
We report here the optimum condition of enzyme production and characterization of crude extract of amylase against the effect of pH, temperature and metal ions addition on the activity of the amylase enzyme
Summary
Starch molecules are polymers of alpha-D-glucopyranose which will be broken down by amylase enzyme at alpha-1,4 and alpha-l,6-glycosides bonds [1]. This polymer is very abundant after cellulose, found in many sago, cassava, maize, millet, etc. Enzymes derived from termofil microbe and hipertermofil are more widely used in industry, especially industries that use high temperatures in the process. This happens because an enzyme derived from the microbe has thermo stability and optimum activity remains at a high temperature [3]. Microbes producing enzymes that are widely used in industry, namely fungi and bacteria [4]
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