Dos Santos José Diogo Neves, João Susana Afonso, Martín Jesús, Vicente Francisca, Reyes Fernando, Lage Olga Maria
Department of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre S/N, 4169-007 Porto, Portugal.
Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, S/N, 4450-208 Matosinhos, Portugal.
Microorganisms. 2022 Jul 20;10(7):1471. doi: 10.3390/microorganisms10071471.
Oceans hold a stunning number of unique microorganisms, which remain unstudied by culture-dependent methods due to failures in establishing the right conditions for these organisms to grow. In this work, an isolation effort inspired by the iChip was performed using marine sediments from Memoria beach, Portugal. The isolates obtained were identified by 16S rRNA gene analysis, fingerprinted using BOX-PCR and ERIC-PCR, searched for the putative presence of secondary metabolism genes associated with polyketide synthase I (PKS-I) and non-ribosomal peptide synthetases (NRPS), screened for antimicrobial activity against ATCC 25922 and ATCC 29213, and had bioactive extracts dereplicated by LC/HRMS. Of the 158 isolated strains, 96 were affiliated with the phylum , PKS-I and NRPS genes were detected in 53 actinomycetotal strains, and 11 proved to be bioactive (10 against , 1 against and 1 against both pathogens). Further bioactivities were explored using an "one strain many compounds" approach, with six strains showing continued bioactivity and one showing a novel one. Extract dereplication showed the presence of several known bioactive molecules and potential novel ones in the bioactive extracts. These results indicate the use of the bacteria isolated here as sources of new bioactive natural products.
海洋中存在着数量惊人的独特微生物，由于无法为这些生物创造适宜的生长条件，依赖培养的方法尚未对其进行研究。在这项研究中，我们借鉴iChip的原理，对来自葡萄牙Memoria海滩的海洋沉积物进行了分离工作。通过16S rRNA基因分析对获得的分离株进行鉴定，利用BOX-PCR和ERIC-PCR进行指纹图谱分析，搜索与聚酮合酶I（PKS-I）和非核糖体肽合成酶（NRPS）相关的次生代谢基因的假定存在情况，筛选对ATCC 25922和ATCC 29213的抗菌活性，并通过LC/HRMS对生物活性提取物进行去重复分析。在158株分离菌株中，96株属于某一门，在53株放线菌总菌株中检测到了PKS-I和NRPS基因，11株被证明具有生物活性（10株对某一种、1株对另一种以及1株对两种病原体均有活性）。使用“一株多化合物”方法进一步探索了其他生物活性，有6株菌株表现出持续的生物活性，1株表现出一种新的生物活性。提取物去重复分析表明，生物活性提取物中存在几种已知的生物活性分子和潜在的新分子。这些结果表明，此处分离的细菌可作为新型生物活性天然产物的来源。
