EXTRACTION OF AMYLASE FROM FUNGI ISOLATED FROM SOIL OF DHARAN, NEPAL

Abstract

Enzymes are the most important substances which are used today in so many areas like research, in industries and in medical field. There are different types of enzymes prevailing in the nature. Among them, amylases are one of the most significant and are widely used in the industries. Amylases are starch hydrolyzing enzymes that have wide spectrum application in industrial and non-industrial sectors. The aim of the current study was to isolate and identify amylase producing fungi from soil collected from different parts of Dharan, and use in the extraction of enzyme. Amylase is extracted from different sources such as plant, animal and microbes. Amylases are widely extracted from the microorganisms, more specifically from the fungi. Extraction of amylase was done by isolating and identifying fungi from the different soil sample. Soil sample was primarily inoculated in Potato Dextrose Agar containing 1% of the starch. The 3 well developed colonies were further sub cultured into the starch agar medium. The production of amylase was tested by using Gram’s iodine solution, and the one fungal colony was selected for the production of amylase on the basis of maximum zone of clearance after the application of iodine. The selected fungal colony was stained by lacto-phenol cotton blue staining technique and was identified as Aspergillus niger. It was subjected in the fermentation medium and was worked accordingly for the production of enzyme (amylase). The enzyme extracted was amylase, and was verified by pouring on the starch agar medium in which zone of clearance was obtained around the holes containing the enzyme. The amylase enzyme was assayed by using Dinitrosalicylic acid (DNS) method. At the end of DNS method, the absorbance of the crude enzyme was estimated and was found to be 0.03 at 540nm