QUANTITATIVE EXTRACTION AND ESTIMATION OF INTRACELLULAR NICOTINAMIDE NUCLEOTIDES OF ESCHERICHIA-COLI
QUANTITATIVE EXTRACTION AND ESTIMATION OF INTRACELLULAR NICOTINAMIDE NUCLEOTIDES OF ESCHERICHIA-COLI
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DOI：
10.1016/0003-2697(79)90039-3
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发表时间：
1979-01-01
影响因子：
2.9
通讯作者：
TOIVONEN, V
中科院分区：
文献类型：
--
作者：
LILIUS, EM;MULTANEN, VM;TOIVONEN, V
研究了从大肠杆菌中提取烟酰胺核苷酸的条件。大肠杆菌中进行了系统研究。NAD+和NADP+的最适pH值约为1.5，而NADH和NADPH的最适pH值约为12.5。在60度下7分钟。选择C用于提取所有4种核苷酸。向碱性提取物中加入CuCl大大提高了还原形式的回收率。比较了烟酰胺核苷酸的两种灵敏度（10-100 pmol）测定方法。一个分光光度循环方法的基础上减少噻唑蓝形成的测量似乎是更灵敏，更重现性和更好的回收率比荧光循环方法在细菌研究中使用的早期。
The conditions of extraction of nicotinamide nucleotides from E. coli were systematically investigated. The optimum pH for NAD+ and NADP+ appeared to be around 1.5 while that for NADH and NADPH was around 12.5. Seven minutes at 60.degree. C was chosen for the extraction of all 4 nucleotides. Addition of CuCl to the alkaline extract improved considerably the recovery of the reduced forms. Two sensitive (10-100 pmol) assay methods for nicotinamide nucleotides were compared. A spectrophotometric cycling method based on the measurement of reduced thiazolyl blue formation appeared to be more sensitive and more reproducible and gave better recovery than a fluorometric cycling method used earlier in bacterial studies.