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

本申请是用于支持NSERC发现资助项目的高效液相色谱（HPLC）仪器。该研究项目包括鉴定微生物来源的具有生物活性的小分子天然产物。高效液相色谱将用于鉴定和纯化微生物培养物中的小分子代谢物，例如，可溶解无机磷酸盐或产生生物活性磷酸盐。微生物通过产生有机酸来溶解土壤中无机磷酸盐复合体的能力已经被发现了一段时间，但这一过程的生物化学尚未得到很好的理解，并且认为存在其他机制。此外，具有生物活性的膦酸盐小分子已被发现用作除草剂和抗生素，但天然膦酸盐被发现的相对较少，对其微生物合成也知之甚少。为了更多地了解这些过程并识别新分子，来自不同环境的大量DNA将在细菌中表达并筛选诸如磷酸盐增溶和磷酸盐生产等功能。拥有具有所需功能的环境DNA （eDNA）的细菌将被大规模培养并通过HPLC进行表征。这是从微生物培养物中识别小分子并阐明其生物合成途径的关键工具。新的磷酸盐增溶机制的发现将有助于提高不可再生磷肥的有效利用，新的磷酸盐化合物将在农业和医学上产生潜在的影响。该仪器将成为研究计划的重要工具，并将对学生的培训做出重大贡献。

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.