维生素C前体物质2-酮基-L-古龙酸的测定

Vitamin C precursor—determination of 2-keto-L-gulonic acid

目的 有效筛选维生素C前体物质2-酮基-L-古龙酸(2-keto-L-gulonic acid, 2-KLG)生产菌株,鉴定其代谢产物中含有2-酮基-L-古龙酸。方法 通过高效液相色谱法(high performance liquid chromatography, HPLC)测定、薄层层析分析和pH值测定等方法对新型菌株发酵液中的代谢产物2-酮基-L-古龙酸进行鉴定。结果 新型菌株发酵液代谢产物中含有2-酮基-L-古龙酸,其高效液相色谱图保留时间、pH值和薄层层析图谱明显不同于2-酮基-D-葡萄糖酸和5酮基-D-葡萄糖酸,且生化实验初步分析显示该菌属于假单胞菌属。结论 N30-1菌株可利用D-葡糖糖一步发酵产生2-酮基-L-古龙酸,今后通过优化育种可为维生素C的生产提供新的生产方案。

Objective To effectively screen the produced strains of 2-keto-L-gulonic acid(2-KLG) in the precusor substance of vitamin C by identify that its metabolite containing 2-KLG.Methods The 2-KLG,a metabolite in the fermentation broth of novel strains, was identified by high performance liquid chromatography(HPLC),thin layer chromatography and determination on pH value.Results The fermentation metabolites of novel strains contained 2-KLG,the retention time of HPLC,pH value and thin layer chromatography of such novel strains were significantly different from those of 2-DKG and 5-DKG,and the preliminary analysis of biochemical test showed that the novel strains were classified to be Pseudomonas.Conclusion The novel strain N30-1 is capable of producing 2-KLG with D-glucose directly by one stage fermentation, providing a new manufacturing plan for increasing the production of vitamin C by optimized breeding in the future.