Hernández-Alomia Fernanda, Ballesteros Isabel, Castillejo Pablo
Grupo de Investigación en Biodiversidad, Medio Ambiente y Salud (BIOMAS), Universidad de las Américas, Quito, Ecuador.
Departamento de Genética, Fisiología y Microbiología, Facultad de Biología, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Saudi J Biol Sci. 2022 Mar;29(3):1550-1558. doi: 10.1016/j.sjbs.2021.11.013. Epub 2021 Nov 12.
Phosphonate compounds are the basis of many xenobiotic pollutants, such as Glyphosate (N-(phosphonomethyl-glycine). Only procaryotic microorganisms and the lower eukaryotes are capable of phosphonate biodegradation through C-P lyase pathways. Thus, the aim of this study was to determine the presence of C-P lyase genes in Ecuadorian freshwater systems as a first step towards assessing the presence of putative glyphosate degraders. To that end, two Nested PCR assays were designed to target the gene that codifies for the subunit J (), which breaks the C-P bond that is critical for glyphosate mineralization. The assays designed in this study led to the detection of genes in 7 out of 8 tested water bodies. The amplified fragments presented 85-100% sequence similarity with genes that belong to glyphosate-degrading microorganisms. Nine sequences were not reported previously in the GenBank. The presence of phosphonate degraders was confirmed by isolating three strains able to grow using glyphosate as a unique carbon source. According to the 16S sequence, these strains belong to the , and genera. Performing a Nested PCR amplification of genes isolated from eutrophicated water bodies, prior to isolation, may be a cost-effective strategy for the bioprospection of new species and/or genes that might have new properties for biotech industries, laying the groundwork for additional research.
膦酸盐化合物是许多外源生物污染物的基础,例如草甘膦(N-(膦酰基甲基)甘氨酸)。只有原核微生物和低等真核生物能够通过C-P裂解酶途径进行膦酸盐生物降解。因此,本研究的目的是确定厄瓜多尔淡水系统中C-P裂解酶基因的存在情况,作为评估假定草甘膦降解菌存在的第一步。为此,设计了两种巢式PCR检测方法,以靶向编码亚基J()的基因,该亚基可断裂对草甘膦矿化至关重要的C-P键。本研究设计的检测方法在8个测试水体中的7个中检测到了基因。扩增片段与属于草甘膦降解微生物的基因具有85-100%的序列相似性。九个序列以前在GenBank中未被报道。通过分离出三株能够以草甘膦作为唯一碳源生长的菌株,证实了膦酸盐降解菌的存在。根据16S序列,这些菌株属于、和属。在分离之前,对从富营养化水体中分离的基因进行巢式PCR扩增,可能是一种具有成本效益的策略,用于生物勘探可能对生物技术产业具有新特性 的新物种和/或基因,为进一步的研究奠定基础。