Stocker Franz, Obermeier Melanie M, Resch Katharina, Berg Gabriele, Müller Bogotá Christina A
Institute of Environmental Biotechnology, Graz University of Technology, Petersgasse 12/I, 8010 Graz, Austria.
Austrian Centre of Industrial Biotechnology, Petersgasse 14/V, 8010 Graz, Austria.
Antibiotics (Basel). 2020 Oct 22;9(11):726. doi: 10.3390/antibiotics9110726.
The ever-growing spread of resistance in medicine and agriculture highlights the need to identify new antimicrobials. Microbial volatile organic compounds (VOCs) are one of the most promising groups of chemicals to meet this need. These rarely exploited molecules exhibit antimicrobial activity and their high vapour pressure makes them ideal for application in surface sterilisation, and in particular, in biofumigation. Therefore, we adapted the previously developed Two Clamp VOCs Assay (TCVA) to a new high-throughput screening for the detection of novel antifungal VOCs from metagenomic clone libraries. As a proof of concept, we tested the new high-throughput TCVA (htTCVA) by sourcing a moss metagenomic library against . This led to the identification of five clones that inhibited the growth of mycelium and spores in vitro by up to 8% and 30% and subsequently, to the identification of VOCs that are potentially, and in part responsible for the clones' antifungal activity. For these VOCs, the in vitro effect of the pure compounds was as high as 100%. These results demonstrate the robustness and feasibility of the htTCVA, which provides access to completely new and unexplored biosynthetic pathways and their secondary metabolites.
医学和农业中耐药性的不断蔓延凸显了识别新型抗菌药物的必要性。微生物挥发性有机化合物（VOCs）是满足这一需求最具潜力的一类化学物质。这些鲜有开发的分子具有抗菌活性，其高蒸气压使其非常适合用于表面灭菌，尤其是生物熏蒸。因此，我们将先前开发的双夹式VOCs测定法（TCVA）改进为一种新的高通量筛选方法，用于从宏基因组克隆文库中检测新型抗真菌VOCs。作为概念验证，我们通过针对苔藓宏基因组文库筛选，对新的高通量TCVA（htTCVA）进行了测试。这导致鉴定出五个克隆，它们在体外对菌丝体和孢子生长的抑制率分别高达8%和30%，随后又鉴定出可能部分导致这些克隆产生抗真菌活性的VOCs。对于这些VOCs，纯化合物的体外效果高达100%。这些结果证明了htTCVA的稳健性和可行性，它为获取全新且未被探索的生物合成途径及其次生代谢产物提供了途径。
