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首页> 外文期刊>Applied Microbiology >Quantitative Extraction of Adenosine Triphosphate from Cultivable and Host-Grown Microbes: Calculation of Adenosine Triphosphate Pools
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Quantitative Extraction of Adenosine Triphosphate from Cultivable and Host-Grown Microbes: Calculation of Adenosine Triphosphate Pools

机译：从可培养和宿主生长的微生物中定量提取三磷酸腺苷：三磷酸腺苷池的计算

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Existing data on adenosine triphosphate (ATP) pools in microbes are deficient for two reasons: (i) incomplete extractions of ATP, and (ii) the failure to take into account that the adverse effects of extracting procedures on standard ATP exert analogous effects on the ATP released from bacterial cells. Methods for correcting observed yields and calculating ATP pools have been demonstrated. Three bacterial species were used in the studies on extraction of ATP: Escherichia coli, Mycobacterium phlei, and Mycobacterium lepraemurium. Perchloric acid and n-butanol were disqualified because of their failure to extract total bacterial ATP even from E. coli and because of inconvenient procedures. The new extraction procedure had minimal effects on standard ATP, liberated 100% of the ATP pools from the three representative species of microbes, and caused no ionic imbalance or quenching of bioluminescence. This method involves vortexing of cell suspensions for 10 s with 23% chloroform (vol/vol), heating at 98 C for the required time (E. coli, 3 min; M. phlei, 5 min; M. lepraemurium, 10 min) and then 1 min at 98 C with vacuum to dry the samples. Heat or chloroform alone may suffice for some microbes and release total ATP from plant and animal cells.
机译：有关微生物中三磷酸腺苷（ATP）库的现有数据不足，其原因有两个：（i）ATP提取不完全;（ii）无法考虑到提取程序对标准ATP的不利影响会导致类似的影响。 ATP从细菌细胞释放。已经证明了校正观察到的产量和计算ATP库的方法。在ATP的提取研究中使用了三种细菌：大肠杆菌，牛分枝杆菌和麻风分枝杆菌。高氯酸和正丁醇不合格，因为它们甚至不能从大肠杆菌中提取总细菌ATP，并且操作不方便。新的提取程序对标准ATP的影响最小，从三种代表性微生物中释放了100％的ATP库，并且没有引起离子失衡或生物发光的猝灭。此方法涉及用23％的氯仿（体积/体积）涡旋细胞悬液10 s，在98°C加热所需的时间（大肠杆菌，3分钟; phlei，5分钟; lepraemurium，10分钟）。 ），然后在真空下于98 C干燥1分钟以干燥样品。单独加热或氯仿可能足以满足某些微生物的需要，并从动植物细胞中释放出总ATP。

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