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酿酒酵母中无氧和有氧糖酵解的研究。

Studies of anaerobic and aerobic glycolysis in Saccharomyces cerevisiae.

作者信息

den Hollander J A, Ugurbil K, Brown T R, Bednar M, Redfield C, Shulman R G

出版信息

Biochemistry. 1986 Jan 14;25(1):203-11. doi: 10.1021/bi00349a029.

Abstract

Glucose metabolism was followed in suspensions of Saccharomyces cerevisiae by using 13C NMR and 14C radioactive labeling techniques and by Warburg manometer experiments. These experiments were performed for cells grown with various carbon sources in the growth medium, so as to evaluate the effect of catabolite repression. The rate of glucose utilization was most conveniently determined by the 13C NMR experiments, which measured the concentration of [1-13C]glucose, whereas the distribution of end products was determined from the 13C and the 14C experiments. By combining these measurements the flows into the various pathways that contribute to glucose catabolism were estimated, and the effect of oxygen upon glucose catabolism was evaluated. From these measurements, the Pasteur quotient (PQ) for glucose catabolism was calculated to be 2.95 for acetate-grown cells and 1.89 for cells grown on glucose into saturation. The Warburg experiments provided an independent estimate of glucose catabolism. The PQ estimated from Warburg experiments was 2.9 for acetate-grown cells in excellent agreement with the labeled carbon experiments and 4.6 for cells grown into saturation, which did not agree. Possible explanations of these differences are discussed. From these data an estimate is obtained of the net flow through the Embden-Meyerhof-Parnas pathway. The backward flow through fructose-1,6-bisphosphatase (Fru-1,6-P2-ase) was calculated from the "scrambling" of the 13C label of [1-13C]glucose into the C1 and C6 positions of trehalose. Combining these data allowed us to calculate the net flux through phosphofructokinase (PFK). For acetate-grown cells we found that the relative flow through PFK is a factor of 1.7 faster anaerobically than aerobically.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

通过使用¹³C核磁共振和¹⁴C放射性标记技术以及瓦氏呼吸计实验,对酿酒酵母悬浮液中的葡萄糖代谢进行了跟踪研究。这些实验针对在生长培养基中使用各种碳源生长的细胞进行,以评估分解代谢物阻遏的影响。葡萄糖利用速率最方便通过¹³C核磁共振实验来确定,该实验测量了[1-¹³C]葡萄糖的浓度,而终产物的分布则通过¹³C和¹⁴C实验来确定。通过结合这些测量结果,估算了进入各种参与葡萄糖分解代谢途径的流量,并评估了氧气对葡萄糖分解代谢的影响。根据这些测量结果,计算出乙酸盐培养的细胞葡萄糖分解代谢的巴斯德商(PQ)为2.95,葡萄糖培养至饱和的细胞为1.89。瓦氏实验提供了葡萄糖分解代谢的独立估算值。从瓦氏实验估算的乙酸盐培养细胞的PQ为2.9,与标记碳实验结果非常吻合,而培养至饱和的细胞的PQ为4.6,两者不一致。讨论了这些差异的可能解释。根据这些数据估算了通过糖酵解途径的净流量。通过[1-¹³C]葡萄糖的¹³C标记“混排”到海藻糖的C1和C6位置来计算通过果糖-1,6-二磷酸酶(Fru-1,6-P2-ase)的逆向流量。结合这些数据使我们能够计算通过磷酸果糖激酶(PFK)的净通量。对于乙酸盐培养的细胞,我们发现厌氧条件下通过PFK的相对流量比需氧条件下快1.7倍。(摘要截选至250字)

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