Harrison D E
J Cell Biol. 1970 Jun;45(3):514-21. doi: 10.1083/jcb.45.3.514.
Klebsiella aerogenes was grown in chemostat culture with the pH controlled to +/-0.01 and temperature to +/-0.1 degrees C. The oxygen tension of the culture was regulated by changing the partial pressure of oxygen in the gas phase and recorded by means of an oxygen electrode. Reduced pyridine nucleotide was monitored continuously in the culture by means of direct fluorimetry. On applying an anaerobic shock to the culture, damped oscillations in pyridine nucleotide fluorescence were obtained. Further anaerobic shocks decreased the damping and eventually gave rise to undamped oscillations of a 2-3 min period which continued for several days. These oscillations were paralleled by oscillations of the same frequency in respiration rate. The amplitude of the oscillations in the respiration rate was equivalent to only 1% of the total steady-state respiration, whereas that of pyridine nucleotide oscillations was equivalent to 10% of the total aerobic/anaerobic fluorescence response. The oscillations ceased on interrupting the glucose feed but restarted on adding excess glucose to the culture. Addition of succinate also restarted the oscillations so that they appear not to be of glycolytic origin. The frequency of oscillations varied with growth rate and conditions. Oscillations of much lower frequency were obtained under limited-oxygen and anaerobic conditions than under fully aerobic conditions. Under glucose-limited conditions, fluctuations were found in adenosine triphosphate (ATP) content which were in phase with the pyridine nucleotide oscillations, but under nitrogen-limited growth conditions no such fluctuations in ATP were observed. The primary oscillating pathway could not be identified but the mechanism would appear to be quite different from that involved in oscillations observed in yeast cells. The synchronization of oscillations and observations of negative damping could be explained by a syntalysis effect.
产气克雷伯菌在恒化器培养中生长,pH控制在±0.01,温度控制在±0.1摄氏度。通过改变气相中的氧分压来调节培养物的氧张力,并通过氧电极进行记录。通过直接荧光法连续监测培养物中还原型吡啶核苷酸。对培养物施加厌氧冲击后,获得了吡啶核苷酸荧光的阻尼振荡。进一步的厌氧冲击降低了阻尼,最终产生了持续2 - 3分钟的无阻尼振荡,这种振荡持续了几天。这些振荡与呼吸速率中相同频率的振荡平行。呼吸速率振荡的幅度仅相当于总稳态呼吸的1%,而吡啶核苷酸振荡的幅度相当于总需氧/厌氧荧光响应的10%。中断葡萄糖进料后振荡停止,但向培养物中添加过量葡萄糖后振荡重新开始。添加琥珀酸也会使振荡重新开始,因此它们似乎不是糖酵解起源的。振荡频率随生长速率和条件而变化。在有限氧和厌氧条件下获得的振荡频率比在完全需氧条件下低得多。在葡萄糖限制条件下,发现三磷酸腺苷(ATP)含量的波动与吡啶核苷酸振荡同步,但在氮限制生长条件下未观察到ATP的这种波动。无法确定主要的振荡途径,但该机制似乎与酵母细胞中观察到的振荡机制有很大不同。振荡的同步和负阻尼的观察可以用合成效应来解释。