植物内生菌来源的生理活性物质的筛选

【作者】 吴芳婷；

【导师】 钱秀萍；

【作者基本信息】 上海师范大学 ， 微生物学， 2004， 硕士

【摘要】 本研究对1760份植物内生菌发酵样品的活性通过多个模型进行了初步筛选，结果发现多个活性样品，对活性样品进行复筛，发现其中有三份样品对巨噬细胞泡沫化抑制活性仍较强。因此对这3株活性菌株的培养特征及生理生化特征进行了研究并对其进行了初步归类。 对具有巨噬细胞泡沫化抑制活性的植物内生菌HCCB00017的代谢产物进行研究，应用溶媒萃取、硅胶柱分离等方法，从其发酵液中分离出具有细胞毒性的活性物质HCCB00017-A，以及具有巨噬细胞泡沫化抑制活性的组分HCCB00017-C和HCCB00017-E。对HCCB00017-A进行质谱和紫外光谱的分析，以及理化性质的初步确定。 本研究建立了针对耐药铜绿假单孢菌(Pseudomonas Aeruginosa即PA)的抗菌增效剂筛选模型。通过对紫外线诱变条件的优化，最终得到了合适的诱变条件。应用此条件经过不断筛选，最后获得对多种抗生素耐药的突变菌株PA9911-C64。药物敏感性试验表明该菌对庆大霉素、链霉素、头孢他啶、环丙沙星及亚胺培南均耐药，其耐药性在无药培养基上传代10代后无明显变化。以此耐药突变菌株作为模型，对800份植物内生菌所产生的发酵样品进行了初步筛选，并获得了几份活性样品。更多还原

【Abstract】 Endophyte is an exceptionally rich, diverse and novel source for bioactive products screening. 1760 samples produced by endophytes reserved by Shanghai Health Creation Center for Biopharmaceutics R&D were tested on several models, and many bioactive samples were selected. The result of second round indepth-screening for these bioactive samples demonstrated that three samples still possecced great inhibitory ability against foam cell formation process of macrophage. The cultural and physiological characters of these strains were studied.The metabolites eliciting inhibition to foam cell formation process of macrophage produced by endophyte HCCB00017 were studied. Several products were isolated through solvent extraction, and silica gel chromatography et al. One compound, HCCB00017-A, showed cytotoxicity; the other two, HCCB00017-C and HCCB00017-E, showed inhibitory activity against foam cell formation process of macrophage. Physical and chemical characteristics and molecular weight of HCCB00017-A were determined by means of MS, UV methods and physical-chemical analysis.An antibiotic synergist model based on drug-resistant strain Pseudomonas Aeruginosa was established. Using optimum ultraviolet radiation treatment, mutatant strain PA9911-C64 was obtained. Antimicrobial agents susceptibility of this strain illustrated that it was resistant to gentamicin, streptomycin, ciprofloxacin, ceftazidime and imipinem. After ten generations growing on drug-free plate, the resistance of PA9911-C64 kept unchanged. 800 broth samples of endophytes were tested on this model, among which several samples showed antimicrobial or synergic activity.更多还原