Abstract:
A useful tool for the screening of fungi producing biologically active secondary metabolites such as antibiotics and cytotoxic substances has been developed. An agar plate-organic solvent interface cultivation (A/S-IFC) system, which comprised a hydrophobic organic solvent (upper phase) , a fungal mat (middle phase) and an agar plate (lower phase) , was constructed. The metabolite profiles were compared among the A/S-IFC, a traditional submerged cultivation (SmC) and an extractive liquid surface immobilization (Ext-LSI) system consisted of a hydrophobic solvent (upper phase) , a fungal cells-ballooned microspheres (middle phase) and a liquid medium (lower phase) , with high-performance liquid chromatography-photodiode array detector (HPLC-PDA) . In the A/S-IFC, many hydrophobic metabolites vastly different from those in the SmC were accumulated in the organic phase as with the Ext-LSI. For example, a valuable azaphilone, sclerotiorin, was remarkably produced into the organic phase in the A/S-IFC. The A/S-IFC was applied to the screening of antibiotic-producing fungi. As a result of paper disk method, it was found that 321 isolated among 811 strains produced antifungal metabolites (hit rate, 39.6%) . Furthermore, 8, 23, and 30 strains also produced cytotoxic metabolites against SKOV-3 (human ovary adenocarcinoma) , MESO-1 (human malignant pleural mesothelioma) , and Jurkat cells (immortalized human T lymphocyte) .
摘要:
已经开发了用于筛选产生生物活性次级代谢产物如抗生素和细胞毒性物质的真菌的有用工具。琼脂平板-有机溶剂界面培养(A/S-IFC)系统,其中包含疏水性有机溶剂(上相),一个真菌垫(中间相)和一个琼脂板(下相),是建造的。在A/S-IFC之间比较了代谢物谱,传统的浸没培养(SmC)和萃取液体表面固定(Ext-LSI)系统由疏水溶剂(上相)组成,真菌细胞膨胀的微球(中间相)和液体培养基(下层相),高效液相色谱-光电二极管阵列检测器(HPLC-PDA)。在A/S-IFC中,与Ext-LSI一样,许多与SmC中截然不同的疏水性代谢物在有机相中积累。例如,一种有价值的氮杂菲酮,硬化因子,在A/S-IFC中显著地产生到有机相中。将A/S-IFC应用于产抗生素真菌的筛选。由于纸盘方法,发现811株中分离出的321株产生了抗真菌代谢产物(命中率,39.6%)。此外,8,23和30株还产生了针对SKOV-3(人卵巢腺癌)的细胞毒性代谢产物,MESO-1(人类恶性胸膜间皮瘤),和Jurkat细胞(永生化人T淋巴细胞)。