目的：以麻黄碱为目标产物, 探讨氮离子(N+)注入介导麻黄基因组DNA在酵母菌中的转化。方法：通过N+注入介导蓝麻黄基因组DNA分别在酿酒酵母Saccharomyces cerevisiae和异常汉逊酵母Hansenula anomala中随机转化, 转化后的酵母菌经BTB指示性辅助筛选、斜面传代和液体培养后, 用铜铬盐定性检识和RP-HPLC方法, 检测重组酵母菌胞外和胞内麻黄碱的量。结果：获得了遗传稳定的以葡萄糖为碳源、NaNO3为氮源生物合成麻黄碱和(或)伪麻黄碱的重组酵母菌9株。液体培养72 h, RP-HPLC测试胞外麻黄碱和伪麻黄碱的最高量分别为18.85和2.88 mg/L;胞内麻黄碱和伪麻黄碱最高量分别为4.29和22.16 mg/g干细胞。结论：采用离子注入介导麻黄基因组DNA大分子转化技术, 通过适当的筛选方法, 可获得易于人工培养的产生麻黄碱等次生代谢产物的微生物工程菌株。

Objective: To investigate Ephedra genomic DNA transformation into yeasts by nitrogen ion implantation with the sole object of ephedrine products. Methods: The genomic DNA from Ephedra glauca was randomly transferred in Saccharomyces cerevisiae and Hansenula anomala, respectively by nitrogenion bombardment. The recombined yeasts were screened and the content of ephedrine in ecto-and endo-recombined yeasts was determined by the methods of bromothymol blue(BTB)indicator selection. slant cultivation, liquid culture, copper chromic saltqualitative test, and RP-HPLC determination. Results: Nine recombined yeast strains that produce l-ephedrine andd-pseudoephedrinewere obtained, which could useglucose as a carbon source, NaN03 asnitrogensource and begeneticallystable. After cultivated in liquidmedium for 72 h and analyzed by RP-HPLC, the recombined strains could produce extracellular l-ephedrine 18. 85mg/Landd-pseudoephedrine 2. 88 mg/L, intracellular l-ephedrine 4. 29 mg/gand d-pseudoephedrine 22. 16 mg/g dry cell. Conclusion: The results show that the ion implantation mediated Ephedra genomic DNA transformation into yeast could be used and the recombined yeasts trains could be obtained by appropriate screening methods.

国家自然科学基金资助项目(10365001)