摘 要 ：对从广东乳源县的南方红豆杉(Taxus chinensis var. mairei)内生真菌中分离和筛选产紫杉醇的内生真菌进行了研究。从南方红豆杉的树皮、茎部、叶片及叶片研磨物中分离纯化了145株内生真菌,对其中的53株内生真菌采用摇瓶发酵培养的方法筛选产紫杉醇的内生真菌。发酵物和菌丝体经研磨、离心、乙酸乙酯萃取和浓缩,经硅胶薄层层析(TLC),高效液相色谱(HPLC)以及液相色谱-质谱(HPLC-MS)分析和检测,结果表明,从茎部分离的1株内生真菌能够产紫杉醇或其异构体, 产量达180 μg L-1。通过对产紫杉醇内生真菌进行诱变、筛选以及优化培养条件等措施,大规模培养生产紫杉醇是具有可行性的。
Abstract:The isolation and screening pacilitaxel-producing endofungus from Taxus chinensis var. mairei were studied, which derived from Ruyuan Xian, Guangdong Province. The 145 endophytic fungi were obtained from the bark, stem, leaf and leaf grinding juice of T. chinensis var. mairei, respectively, in which 53 strains were fermented by shake flask for screening paclitaxel-producing endophytic fungi. The fermented solutions and thalli were grinded, centrifugated, extracted by ethyl acetate and concentrated, and one strain of endophytic fungal from the shem of T. chinensis var. mairei was identified by using thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and HPLC-mass spectrometry (MS). The strain of endofungus could produce paclitaxel or its isimer with yield of 180 μg L-1. Thus, It should be possible for increasing the yield of paclitaxel on a large scale by mutagenesis, screening and optimizing the culture condition.