Silva-Salinas Anaid, Rodríguez-Delgado Melissa, Gómez-Treviño Jesús, López-Chuken Ulrico, Olvera-Carranza Clarita, Blanco-Gámez Edgar Allan
Centro de Investigación en Biotecnología y Nanotecnología (CIByN), Facultad de Ciencias Químicas, Universidad Autónoma de Nuevo León, Parquede Investigación e Innovación Tecnológica, Km. 10 Autopista al Aeropuerto Internacional Mariano Escobedo, Apodaca C.P. 66629, Nuevo León, Mexico.
Laboratorio de Biología Molecular, CELAES, Facultad de Ciencias Químicas, Universidad Autónoma de Nuevo León, San Nicolás de los Garza C.P. 66455, Nuevo León, Mexico.
Microorganisms. 2021 Sep 1;9(9):1857. doi: 10.3390/microorganisms9091857.
This study analyzed the thermostability and effect of calcium ions on the enzymatic activity of α-amylase produced by strain LB04 isolated from Espinazo Hot springs in Nuevo Leon, Mexico. The enzyme was immobilized by entrapment on agar-agarose beads, with an entrapment yield of 19.9%. The identification of the bacteria was carried out using 16s rDNA sequencing. The enzyme was purified through ion exchange chromatography (IEX) in a DEAE-Sephadex column, revealing a protein with a molecular weight of ≈130 kDa. The enzyme was stable at pH 3.0 and heat stable up to 80 °C. However, the optimum conditions were reached at 65 °C and pH 3.0, with a specific activity of 1851.7 U mg ± 1.3. The agar-agarose immobilized α-amylase had a hydrolytic activity nearly 25% higher when compared to the free enzyme. This study provides critical information for the understanding of the enzymatic profile of strain LB04 and the potential application of the microorganisms at an industrial level, specifically in the food industry.
本研究分析了从墨西哥新莱昂州埃斯皮纳佐温泉分离出的LB04菌株所产α-淀粉酶的热稳定性以及钙离子对其酶活性的影响。该酶通过包埋法固定在琼脂糖珠上，包埋率为19.9%。使用16s rDNA测序对细菌进行鉴定。该酶通过在DEAE-葡聚糖凝胶柱上进行离子交换色谱法（IEX）纯化，结果显示为一种分子量约为130 kDa的蛋白质。该酶在pH 3.0时稳定，热稳定性高达80°C。然而，在65°C和pH 3.0时达到最佳条件，比活性为1851.7 U mg±1.3。与游离酶相比，琼脂糖固定化α-淀粉酶的水解活性高出近25%。本研究为了解LB04菌株的酶谱以及该微生物在工业水平上的潜在应用，特别是在食品工业中的应用提供了关键信息。
