title: Purification and characterization of thermostable α-amylase from thermophilic Anoxybacillus flavithermus.
abstract: This study reports on the purification and characterization of thermostable α-amylase (α-1-4 D-glucan glucanohydrolase EC 3.2.1.1) from a newly isolated Anoxybacillus flavithermus. A. flavithermus was used, which was isolated from hot water springs of Ömer, Afyonkarahisar, Turkey. The gram-positive, spore-forming, motile, moderately thermophilic bacteria were found to be a strain of A. flavithermus analysed by 16S rRNA comparison. The optimal conditions for bacterial growth were determined to be at 20 thh, 55 °C and pH 6.0. Maximum α-amylase activity was obtained at 55 °C at pH 7.0 after 24h of incubation. Thermostable α-amylase from A. flavithermus was purified by 70% (NH4)2SO4 and ion-exchange chromatography (5.2-fold; 65.8% yield). The molecular weight of α-amylase was 60 kDa, as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The α-amylase hydrolyzed soluble starch at 55 °C with Km: 0.005 mM and Vmax: 3.5 μmol min(-1).
authors: Department of Biology, Faculty of Science, Dicle University, 21280 Diyarbakir, Turkey. Electronic address: semaguloglu@hotmail.com.
authors: S
authors: S
authors: Agüloğlu Fincan
authors: Department of Biology, Faculty of Science, Dicle University, 21280 Diyarbakir, Turkey.
authors: B
authors: B
authors: Enez
authors: Department of Biology, Faculty of Arts and Science, Siirt University, 56100 Siirt, Turkey.
authors: S
authors: S
authors: Özdemir
authors: Department of Biology, Faculty of Science, Dicle University, 21280 Diyarbakir, Turkey.
authors: F
authors: F
authors: Matpan Bekler
mesh_terms: D058677:Anoxybacillus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005798:Genes, Bacterial; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D000516:alpha-Amylases / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D012336:RNA, Ribosomal, 16S; D000516:alpha-Amylases
keywords: Anoxybacillus flavithermus; Purification; Thermostable; α-Amylase