摘要
目的从珍稀药用植物麻黄中分离内生真菌,检测其麻黄类生物碱产生情况,并探索最优发酵条件。方法处理麻黄草质茎并分离内生真菌,碘化铋钾试剂初检生物碱,以麻黄碱、伪麻黄碱、甲基麻黄碱为对照采用HPLC法筛选产生麻黄类生物碱的菌株,以HPLC法检测总麻黄类生物碱含量并以其为评价指标筛选出碳源及氮源种类后,以碳源量、氮源量、p H值、温度、发酵时间5个考察因素设计L16(45)进行正交试验,从而确定最佳发酵培养基和发酵条件。结果麻黄草质茎中的内生真菌Es2发酵后菌丝体提取物含有麻黄碱、伪麻黄碱及甲基麻黄碱;Es3发酵后菌丝体提取物含有伪麻黄碱。乳糖和硫酸铵是Es2发酵产麻黄类生物碱最适宜的碳源和氮源,当碳源30 g/L,氮源4 g/L,p H5.0,温度30℃,发酵8 d时,Es产麻黄类生物碱的产量最高。结论采用微生物发酵的方法实现麻黄类生物碱的工业化生产,可缓解麻黄碱市场需求对环境的压力。
Objective To isolate endophytic fungi from the rare medicinal plants ephedra, detect the production of ephedrine alkaloids, and explore the optimum fermentation conditions.Methods The endophytic fungi were isolated from the herbaceous stems treated.The ephedrine alkaloids were preliminarily detected by Dragendorff''s reagent.The fungi strains producing ephedrine alkaloids were screened by HPLC method with reference substance of ephedrine, pseudoephedrine and methephedrine.After the optimum carbon source and nitrogen source were determined with total ephedrine alkaloids as evaluation indicators by HPLC, the orthogonal test of L16 (45) was designed to investigate five levels of carbon source amount, nitrogen source amount, pH value, temperature, fermentation time.Results The ephedra herbaceous stems endophytic fungus-Es2 mycelium extract after fermentation contained ephedrine, pseudoephedrine and methephedrine.Es3 mycelium extract containing pseudoephedrine after fermentation.Lactose and ammonium sulfate were as the most suitable carbon source and nitrogen source of Es2 fermentation to produce ephedrine alkaloids.Es produced the highest yield of ephedra alkaloids when the carbon source 30 g/L, nitrogen source 4 g/L, pH5.0, temperature 30℃, fermented for 8 days.Conclusion The ephedrine alkaloids of industrial production could be realized by microorganism fermentation method, which has very important significance to alleviate the ephedrine market demand pressure on the environment.
出处
《中国生化药物杂志》
CAS
2015年第10期18-21,共4页
Chinese Journal of Biochemical Pharmaceutics
关键词
麻黄
内生真菌
麻黄类生物碱
ephedra
endophytic fungi
ephedra alkaloids
