High Performance Liquid Chromatography (HPLC) System for Microbial Natural Products Analysis

本申请是用于支持NSERC发现资助计划的高效液相色谱（HPLC）仪器。该研究计划涉及鉴定微生物来源的生物活性小分子天然产物。HPLC将用于鉴定和纯化微生物培养物中的小分子代谢物，例如，溶解无机磷酸盐或产生生物活性膦酸盐。微生物通过产生有机酸来溶解土壤中无机磷酸盐复合物的能力已经知道了一段时间，但是这个过程的生物化学还没有很好地理解，并且认为存在其他机制。此外，具有生物活性的膦酸酯小分子已被发现用作除草剂和抗生素，但相对较少的天然膦酸酯已被发现，并且对其微生物生物合成的了解甚少。为了更多地了解这些过程并识别新分子，将在细菌中表达来自不同环境的大量DNA，并筛选其功能，如磷酸盐溶解和膦酸盐生产。具有能够实现所需功能的环境DNA（eDNA）的细菌将以更大规模培养并通过HPLC表征。这是从微生物培养物中鉴定小分子和阐明其生物合成途径的关键工具。新的磷增溶机制的发现将有助于更有效地利用不可再生磷肥，新的膦酸酯化合物将在农业和医药领域产生潜在的影响。该仪器将是研究计划的重要工具，并将大大有助于学生的培训。

This application is for a high performance liquid chromatography (HPLC) instrument to be used in support of an NSERC Discovery Grant program. The research program involves identifying biologically active small molecule natural products of microbial origin. The HPLC will be used to identify and purify small molecule metabolites from cultures of microorganisms that, for example, solubilize inorganic phosphate or produce biologically active phosphonates. The ability of microorganisms to solubilize inorganic phosphate complexes in soil via production of organic acids has been known for some time, but the biochemistry of this process is not well understood and other mechanisms are thought to exist. In addition, phosphonate small molecules possessing biological activity have found use as herbicides and antibiotics, yet relatively few natural phosphonates have been discovered and little is understood about their microbial biosynthesis. In order to learn more about these processes and identify novel molecules, massive collections of DNA from different environments will be expressed in bacteria and screened for functions such as phosphate solubilization and phosphonate production. The bacteria possessing the environmental DNA (eDNA) capable of the desired function will be cultured on a larger scale and characterized by HPLC. This is a crucial instrument for identifying small molecules from microbial cultures and for elucidating their biosynthetic pathways. Identification of novel mechanisms of phosphate solubilization will contribute to more efficient use of non-renewable phosphate fertilizer, and new phosphonate compounds will have potential impact in agriculture and medicine. The instrument will be an essential tool for the research program and will contribute significantly to student training.