ISSN 2736-1756
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 12 (11), pp. 001-008, November, 2018. © International Scholars Journals
Full Length Research Paper
Purification and characterization of the α-amylase isolated from PENICILLIUM CAMEMBERTI PL21
Tahar Nouadri*, Zahia Meraihi, Djekrif-Dakhmouche Shahrazed and Bennamoun Leila
Research Laboratory in Enzymatic Engineering, Department of Biochemistry and Microbiology, Mentouri- University, Constantine- Algeria.
Accepted 11 June, 2018
Abstract
The amylase family of enzymes is a great significance due to its wide area of potential application. α-amylase from PENICILLIUM CAMEMBERTI PL21 obtained from I. N. A. (The Technological Laboratory of Agriculture National Institute Paris- France) and using orange waste as substrates was produced under optimum conditions, after 168 h of incubation and subjected for purification and characterization. The enzyme was purified by ammonium sulfate precipitation, dialysis, sephadex G-100 and DEAE-Sepharose CL-6B column chromatography. A trial for the purification of α-amylase resulted in an enzyme with specific activity of (154.2 units/ml/mg protein) with (38.5 folds) purification .The α-amylase activity increased by enzyme concentration rise. The optimum substrate concentration for soluble starch was 1 % (w/v) while the optimum incubation temperature was 30ºC. The purified α-amylase enzyme had a maximum activity at pH 6 and the Km value for soluble starch was 0.92 mg/ml. Analyses of this enzyme for molecular mass was carried out by SDS-PAGE electrophoresis, which revealed one band 60.5 Kda.
Key words: Penicillium camemberti PL21, α-amylase, purification, SDS-PAGE electrophoresis.