Siegel L S, Hylemon P B, Phibbs P V
J Bacteriol. 1977 Jan;129(1):87-96. doi: 10.1128/jb.129.1.87-96.1977.
A modified Gilman assay was used to determine the concentrations of cyclic adenosine 3',5'-monophosphate (cAMP) in rapidly filtered cells and in the culture filtrates of Pseudomonas aeruginosa, Escherichia coli K-12, and Bacteroides fragilis. In P. aeruginosa cultures, levels of cAMP in the filtrate increased with the culture absorbance (3.5 to 19.8 X 10(-9) M) but did not vary significantly with the carbon source used to support growth. Intracellular concentrations (0.8 to 3.2 X 10(-5) M) were substantially higher and did not vary appreciably during growth or with carbon source. Sodium cAMP (5 mM) failed to reverse the catabolite repression of inducible glucose-6-phosphate dehydrogenase (EC 1.1.1.49) synthesis caused by the addition of 10 mM succinate. Exogenous cAMP also had no discernible effect on the catabolite repression control of inducible mannitol dehydrogenase (EC 1.1.1.67). P. aeruginosa was found to contain both soluble cAMP phosphodiesterase (EC 3.1.4.17) and membrane-associated adenylate cyclase (EC 4.6.1.1) activity, and these were compared to the activities detected in crude extracts of E. coli. B. fragilis crude cell extracts contain neither of these enzyme activities, and little or no cAMP was detected in cells or culture filtrates of this anaerobic bacterium.
采用改良的吉尔曼试验法,测定了铜绿假单胞菌、大肠杆菌K-12和脆弱拟杆菌快速过滤细胞及培养滤液中3',5'-环磷酸腺苷(cAMP)的浓度。在铜绿假单胞菌培养物中,滤液中cAMP水平随培养物吸光度增加(3.5至19.8×10⁻⁹ M),但随用于支持生长的碳源变化不显著。细胞内浓度(0.8至3.2×10⁻⁵ M)显著更高,在生长过程中或随碳源变化不明显。5 mM的cAMP钠盐未能逆转因添加10 mM琥珀酸盐而导致的诱导型葡萄糖-6-磷酸脱氢酶(EC 1.1.1.49)合成的分解代谢物阻遏。外源性cAMP对诱导型甘露醇脱氢酶(EC 1.1.1.67)的分解代谢物阻遏控制也没有明显影响。发现铜绿假单胞菌同时含有可溶性cAMP磷酸二酯酶(EC 3.1.4.17)和膜相关腺苷酸环化酶(EC 4.6.1.1)活性,并将这些活性与大肠杆菌粗提物中检测到的活性进行了比较。脆弱拟杆菌粗细胞提取物中不含有这些酶活性,在这种厌氧菌细胞或培养滤液中几乎检测不到cAMP。