The investigation of the production, characterization and biotechnological usage possibilities of the acidic, alkaline, thermostable amylase and pullulanase enzymes isolated from Bacillus sp.

In this study, the production and characterization of an alkaline amylase from neutral strain Aeribacillus pallidus NA1, an acidic amylase from acidic strain Geobacillus lituanicus NA2, an acidic pullulanase from neutral strain Geobacillus thermoleovorans NP1 and an alkaline pullulanase from acidic Anoxybacillus rupiensis NP2 isolated from soil were performed. For this purpose, the analyzes of optimum temperature and pH values, the temperature and pH stabilities, and the effects of various chemicals on enzyme activities were determined, the SDS-PAGE/zymogram and thin layer chromatography (TLC) analyzes were done. The optimum pH values of NA1, NA2, NP1, and NP2 are 9.0, 4.0, 6.0 ve 9.0, respectively. The optimum temperature values of the enzymes are 50, 60, 50, and 60ºC, respectively. And all the enzymes are thermophilic. NA2 acidic amylase and NP2 alkaline pullulanase hydrolyzed starch and pullulan to glucose, maltose and maltotriose, as final products. The enzymes are α-amylase and amylopullulanase. All the enzymes liquefied the starch at an average of 90% up to 30% concentration of starch. And 19% BriX value was obtained by the liquefaction of the starch by NA1 alkaline amylase. The molecular weight of NA1 is 165 kDa, NA2 is 200 kDa and the NP2 enzyme is 113 kDa in size. The NP1 enzyme showed two activity bands with molecular weights of 112 and 107 kDa in zymogram analysis The mixture of NA2 enzyme and 1% detergent showed 100% performance on the removal of foodstuff, baby food and chocolate stains. The isolated enzymes have a high potential for their usage in a variety of biotechnological and industrial applications, particularly in the liquefaction of starch, bioethanol production and the detergent industry. Key Words: Amylase, Amylopullulanase, Acidic, Alkaline, Thermophilic