Sáez M J, Lagunas R
Mol Cell Biochem. 1976 Nov 30;13(2):73-8. doi: 10.1007/BF01837056.
The effect of sampling conditions on the levels of adenine nucleotides, pyridine nucleotides, glycolytic intermediates and related metabolites in yeast has been studied. A systematic examination of the conditions for harvesting has shown that it can be best accomplished by rapid filtration. Delays in the handling for removal of the medium, as is usual in the process of obtaining a number of data reported in the literature, lead to important changes in some of the metabolites examined. It is also shown that when a washing is imperative it can be carried out with a methanol-water mixture (50/50, v/v) cooled at -40 degrees without loss of intracellular concentrations of non-readily diffusible metabolites. On the basis of this experience the outline of a generally applicable procedure is presented.
研究了取样条件对酵母中腺嘌呤核苷酸、吡啶核苷酸、糖酵解中间产物及相关代谢物水平的影响。对收获条件进行的系统检查表明，通过快速过滤可以最好地完成收获。正如文献中报道的获取大量数据的过程中常见的那样，去除培养基的操作延迟会导致所检测的一些代谢物发生重要变化。还表明，当必须进行洗涤时，可以用冷却至 -40 度的甲醇 - 水混合物（50/50，v/v）进行，而不会损失细胞内不易扩散的代谢物浓度。基于这一经验，给出了一个普遍适用程序的概述。
