ISOLATION OF α-AMYLASE GENE FROM Leucosporidium
antarcticum PI 12
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1
Mohd Akmal Azhar, 2Rosli Md Illias & 1Salehhuddin Hamdan
Faculty of Bioscience & Bioengineering, 2Faculty of Chemical and Natural Resources
Engineering, University Technology Malaysia
Email: malcold85@yahoo.com
Abstract
α-Amylase is one of the most important industrial enzymes which can be used for industrial processes
including brewing, baking, textile & detergent. α-Amylase (α-1,4-glucan-4-glucanohydrolases, EC 3.2.1.1)
catalyse the hydrolysis of α-D(1,4)-glucan linkages in both starch and glycogen. Several amylases from
prokaryotic and eukaryotic organisms have been studied previously, but to date very little is known about
these enzymes from psychrophiles. It showed a broad range of potential and application in industrial,
agricultural and medical processes for the psychrophilic organisms and their products because of their low
temperature advantages. Genomic DNA from yeast Leucosporidium antarcticum PI 12 was extracted and
used as template for amplification of α-Amylase gene. About 1850bp of α-Amylase gene was amplified
using PCR. Full length sequence of α-Amylase was obtained via DNA Walking method by extended
upstream and downstream sequence. About 2650bp fragment with 11 exons and 12 introns was indentified
within the α-Amylase gene. About 66% identity showed for the predicted amino acids sequence with αAmylase Cryptococcus neoformans.
Keywords: Alpha amylase, Leucosporidium antarcticum, DNA Walking, α-1,4-glucan-4glucanohydrolases