Department of Microbiology and Biochemistry, Faculty of Natural and Life Sciences, University of Batna 2, 05078, Batna, Algeria.
Molecular Biology Research Laboratory, Department of Biology, Faculty of Science, Karadeniz Technical University, 61080, Trabzon, Turkey.
Braz J Microbiol. 2020 Dec;51(4):1987-2007. doi: 10.1007/s42770-020-00376-0. Epub 2020 Sep 21.
This study aims to determine the diversity of culturable thermophilic bacteria isolated from eight terrestrial hot springs in Northeastern of Algeria using the conventional methods, SDS-PAGE fingerprinting of whole-cell proteins and 16S rRNA gene sequencing. In addition, their hydrolytic enzyme activities were also investigated. A total of 293 strains were isolated from the hot springs' water and sediment using different culture media. Overall, five distinct bacterial groups were characterized by whole-cell protein pattern analysis. Based on the 16S rRNA gene sequencing of 100 selected strains, the isolates were assigned to the following three major phyla: Firmicutes (93%), Deinococcus-Thermus (5%), and Actinobacteria (2%), which included 27 distinct species belonging to 12 different phylotypes, Aeribacillus, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Geobacillus, Laceyella, Meiothermus, Saccharomonospora, Thermoactinomyces, Thermobifida, and Thermus. The screening for nine extracellular enzymes showed that 65.87% of the isolates presented at least five types of enzyme activities, and 6.48% of strains combined all tested enzymes (amylase, cellulase, pectinase, esculinase, protease, gelatinase, lipase, lecithinase, and nuclease). It was found that Bacillus, Anoxybacillus, Aeribacillus, and Aneurinibacillus were the genera showing the highest activities. Likewise, the study showed an abundant and diverse thermophilic community with novel taxa presenting a promising source of thermozymes with important biotechnological applications. This study showed that a combined identification method using SDS-PAGE profiles of whole-cell proteins and subsequent 16S rRNA gene sequence analysis could successfully differentiate thermophilic bacteria from Algerian hot springs.
本研究旨在使用常规方法、全细胞蛋白 SDS-PAGE 指纹图谱和 16S rRNA 基因测序,从阿尔及利亚东北部的 8 个陆地温泉中分离出可培养嗜热细菌的多样性。此外,还研究了它们的水解酶活性。使用不同的培养基从温泉水和沉积物中分离出了 293 株菌株。总体而言,通过全细胞蛋白图谱分析,将 5 个不同的细菌群进行了特征描述。基于 100 株选定菌株的 16S rRNA 基因测序,将分离株分配到以下三个主要门:厚壁菌门(93%)、无壁菌门-栖热菌门(5%)和放线菌门(2%),其中包括 27 个不同的种,属于 12 个不同的类群,气杆菌属、副球菌属、厌氧芽孢杆菌属、芽孢杆菌属、短芽孢杆菌属、地芽孢杆菌属、莱克氏菌属、嗜热甲基球菌属、喜热放线菌属、嗜热双歧杆菌属和 Thermus。对九种细胞外酶的筛选表明,65.87%的分离株至少具有五种类型的酶活性,6.48%的菌株组合了所有测试的酶(淀粉酶、纤维素酶、果胶酶、七叶苷酶、蛋白酶、明胶酶、脂肪酶、卵磷脂酶和核酸酶)。结果表明,芽孢杆菌属、厌氧芽孢杆菌属、气杆菌属和副球菌属是表现出最高活性的属。同样,该研究表明,存在一个丰富多样的嗜热菌群,其中有新的类群,是具有重要生物技术应用前景的嗜热酶的有前途的来源。本研究表明,使用 SDS-PAGE 全细胞蛋白图谱和随后的 16S rRNA 基因序列分析相结合的鉴定方法,可以成功地从阿尔及利亚温泉中分离出嗜热细菌。