Laboratorio de Microorganismos Extremófilos, Instituto Antofagasta, Universidad de Antofagasta, Antofagasta 1240000, Chile.
Instituto de Ciencias Naturales Alexander von Humboldt, Facultad de Ciencias del Mar y Recursos Biológicos, Universidad de Antofagasta, Antofagasta 1240000, Chile.
Int J Mol Sci. 2023 Sep 25;24(19):14512. doi: 10.3390/ijms241914512.
The present study describes the isolation of an extremely thermophilic bacterium from El Tatio, a geyser field in the high planes of Northern Chile. The thermophile bacterium named strain ET-1 showed 99% identity with SGO.5JP 17-16 (GenBank accession No. CP002777) by 16S rDNA gene analysis. Morphologically, the cells were non-sporeforming Gram-negative rods that formed colonies with yellow pigmentation. This strain is able to proliferate between 55 and 80 °C with a pH range of 6-10, presenting an optimum growth rate at 80 °C and pH 8. The bacterium produces an extracellular protease activity. Characterization of this activity in a concentrated enzyme preparation revealed that extracellular protease had an optimal enzymatic activity at 80 °C at pH 10, a high thermostability with a half-life at 80 °C of 10 h, indicating that this enzyme can be classified as an alkaline protease. The proteolytic enzyme exhibits great stability towards chelators, divalent ions, organic solvents, and detergents. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF), implying that it was a serine protease. The high thermal and pH stability and the resistance to chelators/detergents suggest that the protease activity from this strain could be of interest in biotechnological applications.
本研究描述了一种从智利北部高原间歇泉地区 El Tatio 分离出的极端嗜热细菌。这种名为 strain ET-1 的嗜热菌,通过 16S rDNA 基因分析,与 SGO.5JP 17-16(GenBank 登录号 CP002777)具有 99%的同源性。形态上,细胞是非孢子形成的革兰氏阴性棒状,形成带有黄色色素的菌落。该菌株能够在 55-80°C 之间增殖,pH 值范围为 6-10,在 80°C 和 pH 值 8 时呈现最佳生长速率。该菌产生细胞外蛋白酶活性。在浓缩酶制剂中对该活性进行的表征表明,细胞外蛋白酶在 pH 值为 10、80°C 时具有最佳酶活性,半衰期在 80°C 时为 10 小时,表明该酶可归类为碱性蛋白酶。该蛋白酶在螯合剂、二价离子、有机溶剂和洗涤剂中具有很强的稳定性。酶被苯甲基磺酰氟 (PMSF) 抑制,表明它是一种丝氨酸蛋白酶。高耐热性和 pH 稳定性以及对螯合剂/洗涤剂的抗性表明,该菌株的蛋白酶活性可能在生物技术应用中具有重要意义。