Quantitative extraction and estimation of intracellular nicotinamide nucleotides of Escherichia coli

✍ Scribed by Esa-Matti Lilius; Veli-Matti Multanen; Veija Toivonen

• Book ID: 102627075
• Publisher: Elsevier Science
• Year: 1979
• Tongue: English
• Weight: 433 KB
• Volume: 99
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(79)90039-3

✦ Synopsis

The conditions of extraction of nicotinamide nucleotides from Escherichia 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°C was chosen for the extraction of all four 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.