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首页> 外文期刊>The Journal of Antibiotics: An International Journal >Identification of aluminium dioxalate in fungal cultures grown on vermiculite
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Identification of aluminium dioxalate in fungal cultures grown on vermiculite

机译：ver石真菌培养物中二草酸铝的鉴定

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Natural products, especially those derived from microorganisms, have had a very important role in human health through the development of new pharmaceutical drugs, crop protection agents, traditional herbal medicines and personal-care products. In order to identify novel natural products, microorganisms continue to be cultured using different growth media to maximise the diversity of secondary metabolite production. In addition to liquid media fermentations, fungi are also grown in solid media, which can mimic the way these organisms grow in nature. One of the solid media used in our fungal fermentations was vermiculite, which is a clay material mined from various locations around the world. Vermiculite is an ideal solid substrate because of its high surface area and ability to absorb water, although maintaining a medium shrink-swell capacity. Fungal fermentations grown on vermiculite media were extracted with MeOH to provide extract libraries that were suitable for high throughput screening (HTS). One of the targets selected for screening was the Mycobacterium tuberculosis-derived enzyme isocitrate lyase (ICL), with isoforms ICL1 and ICL2 being shown to be essential for in vivo growth and virulence. ICLs are the key enzymes used by bacteria, fungi and plants in the glyoxylate cycle that catalyses the cleavage of isocitrate to succinate and glyoxylate. We screened 76466 extracts using a coupled enzyme assay and identified a series of active samples from the vermiculite-based extracts. Active extracts were then subjected to a dereplication procedure; approximately, 1 mg of the crude extract was separated by C18-reversed phase HPLC and collected into 40 fractions in a 96-well microtitre plate. The fractions were dried by centrifugal evaporation and dissolved in 12.5% DMSO/H_2O. Biological testing using the ICL screen revealed that the activity of most of the extracts eluted from the column with the water solvent front.
机译：通过开发新的药物，农作物保护剂，传统草药和个人护理产品，天然产品，尤其是衍生自微生物的天然产品在人类健康中发挥着非常重要的作用。为了鉴定新的天然产物，继续使用不同的生长培养基来培养微生物，以使次级代谢产物产生的多样性最大化。除液体培养基发酵外，真菌还在固体培养基中生长，这可以模仿这些生物在自然环境中的生长方式。 our石是我们真菌发酵中使用的一种固体培养基，ver石是一种从世界各地开采的粘土材料。 maintaining石虽然具有中等的收缩溶胀能力，但由于其高表面积和吸收水的能力，因此是理想的固体基质。在石培养基上生长的真菌发酵液用MeOH提取，以提供适合高通量筛选（HTS）的提取物库。选择用于筛选的靶标之一是结核分枝杆菌衍生的酶异柠檬酸裂合酶（ICL），已显示同工型ICL1和ICL2对于体内生长和毒力必不可少。 ICL是细菌，真菌和植物在乙醛酸循环中使用的关键酶，催化异柠檬酸裂解为琥珀酸和乙醛酸。我们使用偶联酶分析法筛选了76466提取物，并从基于ver石的提取物中鉴定了一系列活性样品。然后将活性提取物进行重复复制程序;通过C18反相HPLC分离大约1mg的粗提取物，并收集在96孔微量滴定板中的40个级分中。将级分通过离心蒸发干燥并溶解在12.5％的DMSO / H_2O中。使用ICL筛分仪进行的生物学测试表明，大多数萃取物的活性均从带有水溶剂前沿的色谱柱上洗脱出来。

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