低丰度^(13)C标记的酿酒酵母代谢通量分析探讨研究

Research on ^(13)CMFA of S.cerevisiae at Low Labeled Tracer Substrate

通过对酿酒酵母发酵液进行低浓度的[1-13C]葡萄糖标记,并采用GC-C-IRMS测定发酵菌体中蛋白氨基酸的δ13C值,最终显示在0.5%~2%的标记浓度范围内,15种菌体蛋白氨基酸δ13C值与标记浓度呈现出很好的线性关系。证明了0.5%或者更低的浓度标记都可以用于13CMFA中,大大降低了碳同位素标记代谢流成本,为13CMFA应用于传统白酒和黄酒大生产发酵过程中的复杂代谢机理研究提供一种手段。

Stable carbon isotope compositions of proteinogenic amino acids in S.cerevisiae fermenting liquid at low labeled tracer substrate were determined by gas chromatography-combustion-isotope ratio mass spectrometry （GC-C-IRMS） in the experiments. The results suggested that, within 0.5 %-2 % labeled range, ^13C values of 15 kinds of amino acids in protein had good linear relationships with the labeled concentration. The experiments demonstrated that 0.5 % or even lower labeled concentration was feasible in ^13CMFA, which could reduce greatly the metabolic flux cost and provide a new approach to ^13CMFA application in the research on the complex metabolism of the fermentation of traditional liquor and yellow rice wine.