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.

摘要

产生嗜热淀粉酶的艾德厌氧杆菌FMB1 是从土耳其的 Sorgun 温泉中分离出来的。为了在成本有效的过程中获得改进的酶生产，需要优化过程参数。使用传统的浸没发酵 (SmF) 方法确定淀粉酶生产的最佳条件。在液体 BMB 培养基中，在优化条件（24 小时、55°C 和 pH 8.0）下，最大淀粉酶活性为 295.8 U/mg。在1.5%可溶性淀粉作为碳源和1%酵母提取物作为氮源存在下获得最佳酶产量。酶在最佳条件下生产并部分纯化。部分纯化的酶在 50 °C 下可稳定长达 120 分钟，发现其稳定性为 pH 5.0–10.0。关于某些金属和化学品对部分纯化酶活性的影响，Ca²⁺使酶活性提高了21%，但酶活性被Mg ²⁺、Zn ²⁺、Ni ²⁺和Cu ^{2+ 分别}降低了5%、29%、57%和64%。Hg ²⁺完全抑制酶活性。此外; DTT、PMSF、PCMB、SDS 和 EDTA 分别导致酶活性降低 3%、4%、12%、27% 和 77%。通过SDS-PAGE发现酶的分子量为58.5 kDa。所得结果表明嗜热A. ayderensis FMB1 及其酶具有生物技术重要性。