Purification and characterization of novel α-amylase from Anoxybacillus ayderensis FMB1

Abstract

The thermophilic amylase producing Anoxybacillus ayderensis FMB1 was isolated from Sorgun Hot Spring in Turkey. In order to obtain improved production of the enzyme for cost-effective process, optimization of the process parameters is required. Optimum conditions for amylase production were determined using the traditional Submerged Fermentation (SmF) method. The maximum amylase activity was 295.8 U/mg, under the optimized conditions (24-h, 55 °C, and pH 8.0) in liquid BMB medium. The best enzyme production was obtained in the presence of 1.5% soluble starch as carbon source and 1% yeast extract as nitrogen source. Enzyme was produced under optimal conditions and partially purified. Partially purified enzyme was stable for up to 120 minutes at 50 °C, and its stability was found to be pH 5.0–10.0. Regarding the effects of some metals and chemicals on the activity of the partially purified enzyme, Ca²⁺ increased the activity by 21%, but enzyme activity was decreased by Mg²⁺, Zn²⁺, Ni²⁺, and Cu²⁺ by 5%, 29%, 57%, and 64%, respectively. and the enzyme activity was completely inhibited by Hg²⁺. In addition; DTT, PMSF, PCMB, SDS and EDTA caused a decrease in enzyme activity by 3%, 4%, 12%, 27% and 77%, respectively. The molecular weight of enzyme was found to be 58.5 kDa by SDS-PAGE. The results obtained show that the thermophilic A. ayderensis FMB1 and their enzyme have biotechnological importance.

Graphical Abstract

Keywords:

• Amylase
• Ca-dependent
• thermostable

Disclosure statement

No potential conflict of interest was reported by the author(s).