Cloning and expression of alpha amylase producing gene from S. aureus into E. coli

Among the top commercial enzymes utilized for industrial applications, amylases are the most widely used and accepted. The aim of present work was to isolate amylase producing bacteria from the soil samples collected from area around the plant roots. Bacterial species were isolated by serial dilution and cultured on the nutrient agar medium plates. The isolated bacteria were evaluated for the amylase activity using starch hydrolysis test. The bacterial isolate with maximum hydrolytic activity was considered for further study. Based on the morphology and biochemical tests, the species was confirmed to be Staphylococcus aureus. Amylase of S. aureus was amplified using PCR with the help of specific primers; and a PCR product of approximately 1.9 kb was obtained. This was cloned into pUC 18 vector which was transformed into competent E. coli DH5? cells. The ability of transformed bacteria to grow on agar plates with ampicillin confirmed successful transformation. Furthermore, these transformed bacteria were able to produced active amylase that is revealed by starch hydrolysis test. Thus S. aureus amylase gene was successfully cloned into E. coli..