金钗石斛内生菌CPU0029的诱变及其次级代谢产物的纯化

Mutagenesis of Endophyte CPU0029 Isolated from Dendribium Nobile Lindl and Purification of Its Secondary Metabolites

以实验室筛选得到的产生物碱的金钗石斛内生菌CPU0029为出发菌株,为了提高生物碱产量对其进行紫外及亚硝酸钠复合诱变,筛选出1株正突变菌株;运用正交实验的方法对发酵液中生物碱成分进行提取优化,最后采用薄层、气相方法进行检测。结果:通过复合诱变筛选出的正突变株,生物碱产量达到6.9mg/L,比出发菌株高出63%,且其遗传性状较稳定;通过提取纯化,该生物碱纯度提高了70.6倍。

To improve the yield and to purity of alkaloids in fermentation broth of Dendribium nobile Lindl endophyte CPUO029 （CCTCC M208020, it had been applied for a patent）, the CPU0029 was taken as original strain and mutated by NaNO2 and UV for breeding high-yield alkaloids mutants. The purification conditions of alkaloids in fermentation broth through PTLC were screened by orthogonal experiments. The final purified sample was detected by TLC and GC. Results showed that one positive mutant strain was selected, whose alkaloids output was 6.9 mg/L,63 % higher than the original strain relatively, and had a stable genetic character. Af- ter extraction and purification,the purity of alkaloids increased 70.6 times. The yield and purity of alkaloids in fermentation broth had improved significantly.