Department of Biology, University of Naples Federico II; Institute of Polymers, Composites and Biomaterials (IPCB), Consiglio Nazionale delle Ricerche CNR.
Department of Biology, University of Naples Federico II; Division of Biological Systems and Engineering, Lawrence Berkeley National Laboratory.
J Vis Exp. 2021 Dec 30(178). doi: 10.3791/63453.
Geothermal springs are rich in various metal ions due to the interaction between rock and water that takes place in the deep aquifer. Moreover, due to seasonality variation in pH and temperature, fluctuation in element composition is periodically observed within these extreme environments, influencing the environmental microbial communities. Extremophilic microorganisms that thrive in volcanic thermal vents have developed resistance mechanisms to handle several metal ions present in the environment, thus taking part to complex metal biogeochemical cycles. Moreover, extremophiles and their products have found an extensive foothold in the market, and this holds true especially for their enzymes. In this context, their characterization is functional to the development of biosystems and bioprocesses for environmental monitoring and bioremediation. To date, the isolation and cultivation under laboratory conditions of extremophilic microorganisms still represent a bottleneck for fully exploiting their biotechnological potential. This work describes a streamlined protocol for the isolation of thermophilic microorganisms from hot springs as well as their genotypical and phenotypical identification through the following steps: (1) Sampling of microorganisms from geothermal sites ("Pisciarelli", a volcanic area of Campi Flegrei in Naples, Italy); (2) Isolation of heavy metal resistant microorganisms; (3) Identification of microbial isolates; (4) Phenotypical characterization of the isolates. The methodologies described in this work might be generally applied also for the isolation of microorganisms from other extreme environments.
热泉富含各种金属离子,这是由于岩石和水在深部含水层相互作用的结果。此外,由于 pH 值和温度的季节性变化,这些极端环境中的元素组成会定期波动,影响环境微生物群落。在火山热液喷口处生长的嗜极微生物已经发展出了应对环境中存在的多种金属离子的抗性机制,从而参与到复杂的金属生物地球化学循环中。此外,嗜极微生物及其产物在市场上找到了广泛的立足点,尤其是它们的酶。在这种情况下,对其进行特性分析对于开发用于环境监测和生物修复的生物系统和生物工艺是功能必需的。迄今为止,在实验室条件下对嗜极微生物进行分离和培养仍然是充分发挥其生物技术潜力的瓶颈。这项工作描述了一种从热泉中分离嗜热微生物的简化方案,以及通过以下步骤对其进行基因和表型鉴定:(1)从地热场(意大利那不勒斯坎皮弗莱格雷的皮斯卡雷利火山地区)采集微生物样品;(2)分离耐重金属的微生物;(3)鉴定微生物分离物;(4)分离物的表型特征分析。本工作中描述的方法通常也可用于从其他极端环境中分离微生物。