重组酵母菌生物合成麻黄碱的特性及L-Phe对其合成的影响

Characteristics of Ephedrine and Pseudoephedrine Biosynthesis in Recombined Yeasts and the Effects of L-Phenylalanine in Alkaloids Biosynthesis

目的:探讨12株重组酵母菌生物合成麻黄碱的特性及L-Phe对麻黄碱生物合成的影响。方法:以葡萄糖为碳源、NaNO3为氮源对重组酵母菌进行培养,在相同条件下,在液体培养基中添加5mg/L的L-Phe,利用反向高压液相色谱(RP-HPLC)测定重组酵母菌培养液中麻黄碱和伪麻黄碱的含量。结果:重组酵母菌培养液中麻黄碱和伪麻黄碱的最高产量分别为18.85mg/L和4.11mg/L;L-Phe对各重组菌株生物合成麻黄碱的调控作用各不相同。结论:通过L-Phe对重组酵母菌生物合成麻黄碱和伪麻黄碱调控作用的探讨,将为研究重组菌株的遗传多样性提供依据。

Objective：Investigation on the characteristics of ephedrine and pseudoephedrine biosynthesis in 12 recombined yeast strains and the effects of L-phenylalanine in alkaloids biosynthesis.Method：Used of glucose as the carbon source,NaNO3 as the nitrogen source to carry on culture to the recombined yeasts,otherwise added the L-Phe of 5mgs/L under the same culture condition.The l-ephedrine and d-pseudoephedrine in the culture liquid of the recombined yeasts were analyzed by the RP-HPLC.Result：After culturing in liquid medium for 72h,extracellular l-ephedrine and d-pseudoephedrine concentrations of 18.85 mg/L and 4.11mg/L,respectively.The addition of the L-Phe to culture medium substantially changed the amount of l-ephedrine and/or d-pseudoephedrine produced by the recombined yeasts.Conclusion：The results of the regulation on L-Phe to the l-ephedrine and d-pseudoephedrine biosynthesis in recombined yeasts would be valuable for further investigations of genetical diversity.