Investigation of the Amylolytic Enzyme Profile of the Bacterium Bacillus amyloliquefaciens BEH 111, and Determination of its α-Amylase Enzyme Characteristics

Aims and Amylases are one of the major industrial enzymes that occupy 25 % of the world enzyme market. These enzymes are used in many different areas، such as paper، textiles، pharmaceutical، food، starch saccharafication and processing industries. Much research has been carried out regarding application of amylolytic enzymes produced by the genus Bacillus، to the production of glucose and beverages، food and detergent industries. Because of the extensive applicability of amylolytic enzymes، research in relation to identification of these enzymes and their characteristic is continuously being carried out. Hence، the aim of this research was to investigate the amylolytic enzyme profile of a native Bacillus strain، and determine its enzyme characteristics. In this study، the enzymatic profile of the bacterium، Bacillus amyloliquefaciens BEH 111، was determined by using the SDS-PAGE، zymogram and TLC methods. Zymography and TLC were used to identify the amylolytic enzyme profile of the bacterium. The molecular weight of α-amylase was estimated by SDS-PAGE. The 3، 5-Dinitrosalisyclic acid (DNS) indicator was used to examine α-amylase activity، quantitatively. Investigation of enzyme characteristics and determination of maximal enzyme activity was carried by the DNS method، at different temperatures، pH values، substrate concentrations and time periods. The molecular weight of α-amylase was estimated to be 55 kDa by SDS-PAGE. Investigation of enzyme characteristics showed that maximum α-amylase activity was achieved at a temperature of 50oC، pH 7، presence of 2% starch، during a 90 min time period. Enzyme activity obtained under these conditions was 8222 U/L. The high level of enzyme activity produced by B. amyloliquefaciens BEH 111 can thus be of commercial significance. Furthermore، by having activity at 50oC and above، makes it possible for this enzyme to be applied to processes that use high temperatures. The optimized pH levels of 6-7 that resulted in high enzyme activity can also allow the potential use of this enzyme in processes such as starch liquefaction and gelatinization.