Purification and Characterization of Extracellular Protease and Amylase Produced by the Bacterial Strain, Corynebacterium alkanolyticum ATH3 Isolated from Fish Gut

The use of enzymes in different industrial sectors increased significantly due to huge industrialization. Different types of protease and amylase are randomly used in industries including food, textile, and paper. For this purpose, purification of extracellular protease and amylase produced by the bacterium, Corynebacterium alkanolyticum ATH3 (Acc. No. JX656749) isolated from the distal intestine of a freshwater fish, Anabas testudineus, was carried out using column chromatography. The specific activity of protease and amylase significantly increased with each step of purification and finally became 93.73 and 88.1 U/mg protein with a purification fold 26.03 and 44.94, respectively. The sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed molecular weight of purified protease and amylase was similar to 17 and similar to 28 kDa, respectively. To the authors' knowledge, this is the first report of similar to 17 kDa protease and similar to 28 kDa amylase from the bacterial strain C. alkanolyticum ATH3. Further, enzyme activity was also evidenced by zymography analysis. The enzymes acted optimally at pH 7.5-8.0 and temperature 35-45 degrees C, respectively. So, due to cheapest source, these two enzymes are very important for various purposes in industrial sectors.