Theobald U, Mailinger W, Reuss M, Rizzi M
Institut für Bioverfahrenstechnik, Universität Stuttgart, Federal Republic of Germany.
Anal Biochem. 1993 Oct;214(1):31-7. doi: 10.1006/abio.1993.1452.
Transition from glucose limitation to glucose excess in Saccharomyces cerevisiae leads to a fast change from complete to partial oxidative metabolism. To determine rapid changes of in vivo concentrations of yeast metabolites, occurring in the range of seconds after a glucose injection, a fast sampling technique was developed. A harvesting device with negligible dead space and highly efficient inactivation and extraction steps was applied to analyze the behavior of the physiological concentrations of adenine nucleotides as the dynamic response to fast changing glucose concentrations. Adenine nucleotides were selected as appropriate references due to their high turnover rates (in the range of seconds). During the first 5 s after a glucose pulse to a continuous chemostat culture, a significant decrease (30%) in ATP concentration and a coincident rise of ADP and AMP values were found. A further advantage of this technique is the high sampling frequency (5 s) and the long-term sterility ensuring recurrent harvesting.
酿酒酵母从葡萄糖限制状态转变为葡萄糖过量状态会导致其从完全氧化代谢迅速转变为部分氧化代谢。为了确定在注射葡萄糖后数秒内酵母代谢物体内浓度的快速变化，开发了一种快速采样技术。采用了一种死体积可忽略不计且具有高效灭活和提取步骤的采集装置，以分析腺嘌呤核苷酸生理浓度的行为，作为对快速变化的葡萄糖浓度的动态响应。由于腺嘌呤核苷酸的高周转率（在数秒范围内），因此被选为合适的参考指标。在向连续恒化器培养物中注入葡萄糖后的最初5秒内，发现ATP浓度显著下降（30%），同时ADP和AMP值相应升高。该技术的另一个优点是采样频率高（5秒）且长期无菌，可确保反复采集。
