Purifi cation and Characterization of Thermostable and Detergent-Stable [alpha]-Amylase from Anoxybacillus sp. AH1

• Published in: Food technology and biotechnology Vol. 54; no. 1; p. 70
• Main Authors: Acer, Ömer, Bekler, Fatma Matpan, Pirinççioglu, Hemse, Güven, Reyhan Gül, Güven, Kemal
• Format: Journal Article
• Language: English
• Published: Zagreb Sveuciliste u Zagrebu, Prehramheno-Biotehnoloski Fakultet 01.01.2016
• Subjects: Acids; Enzymes; Proteins; Studies; Turkey
• ISSN: 1330-9862; 1334-2606
• DOI: 10.17133/ftb.54.01.16.4122

A thermostable and detergent-stable α-amylase from a newly isolated Anoxybacillus sp. AH1 was purifi ed and characterized. Maximum enzyme production (1874.8 U/mL) was obtained at 24 h of incubation. The amylase was purified by using Sephadex G-75 gel filtration, after which an 18-fold increase in specific activity and a yield of 9% were achieved. The molecular mass of the purified enzyme was estimated at 85 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature values of the enzyme were 7.0 and 60 degrees Celsius, respectively. The enzyme was highly stable in the presence of 30% glycerol, retaining 85% of its original activity at 60 degrees Celsius within 120 min. The enzyme activity was increased by various detergents, but it was significantly inhibited in the presence of urea. Mg^sup 2+^ and Ca^sup 2+^ also significantly activated α-amylase, while Zn^sup 2+^, Cu^sup 2+^ and metal ion chelators ethylenediaminetetraacetic acid (EDTA) and 1,10-phenanthroline (phen) greatly inhibited the enzyme activity.