EXTRACTION AND PARTIAL PURIFICATION OF AMYLASE FROM MALUS PUMILA

Rajesh kumar*, Harnek saini and Deepak Kumar Malik * Corresponding author: email: dahiya76@gmail.comDepartment of Biotechnology, University Institute of Engineering & Technology, Kurukshetra University Kurukshetra, Haryana

ABSTRACT

Amylases are starch degrading enzymes that catalyze the hydrolysis of internal alpha 1-4 glycosidic bonds in polysaccharides with retention of alpha anomeric configuration in the products. They are found in all forms of organisms regardless of kingdom. Amylases have attracted attention due to their multitude industrial applications such as, liquefaction, bread making, paper industry, chocolate industry and textile industry. Amylases from plants and microbial sources have been used for centuries in brewing industry. In the present work amylase was isolated from apple (Malus pumila). The enzyme was partially purified by using techniques of ammonium sulphate precipitation and gel filtration. The enzyme has shown initial enzyme activity of .204 IU/ml. After partial purification enzyme showed a purification fold of 24 and yield percentage as 11. The specific activity of enzyme get increased from .204 IU/ml to 4.76 IU/ml. The enzyme was characterized with respect to temperature optima and stability. The enzyme has shown maximum activity at a temperature of 350 C and stability between 30 to 50 0 C up to one hour. The effect of metal ions was also studied. The ca +2, Mn +2 and Mg +2 ions showed positive effects. The enzyme activity was increased by about 36%, 22% and 8% respectively while a significant inhibitory effect on amylase activity was observed with Hg +2. The other metal ions which showed a negative impact included Na+, Cu+2 and Fe+2.