Riendeau D, Meighen E
J Biol Chem. 1980 Dec 25;255(24):12060-5.
The luminescent bacterium Photobacterium phosphoreum has been shown to possess a fatty acid reductase based on the stimulation of the aldehyde-dependent luminescent reaction on incubation of the enzyme with ATP, NADPH, and tetradecanoic acid (Riendeau, D., and Meighen, E. (1979) J. Biol. Chem. 254, 7488-7490). A direct, luciferase-independent assay for the fatty acid reductase has now been developed using [3H]tetradecanoic acid as substrate and thin layer chromatography to separate and identify the products of the reaction. Tritiated aldehyde was the only product of the reaction at early times of assay, and the amount produced was linearly dependent on time and extract concentration. The labeled aldehyde was further reduced to alcohol after prolonged incubation, indicating that long chain aldehyde reductase(s) are also present in bioluminescet bacteria. Measurement of the fatty acid reductase activity in extracts during growth and development of the bioluminescent bacteria showed that the fatty acid reductase activity is co-induced with luciferase, suggesting that these enzymes are coordinately regulated and directly implicating the fatty acid reductase in aldehyde biosynthesis in the bacterial bioluminescent system.
发光细菌费氏弧菌已被证明拥有一种脂肪酸还原酶,这是基于在该酶与ATP、NADPH和十四烷酸一起孵育时,醛依赖性发光反应受到刺激(Riendeau, D., 和Meighen, E. (1979) J. Biol. Chem. 254, 7488 - 7490)。现在已经开发出一种直接的、不依赖荧光素酶的脂肪酸还原酶检测方法,使用[3H]十四烷酸作为底物,并通过薄层色谱法分离和鉴定反应产物。在检测早期,氚化醛是反应的唯一产物,其产生量与时间和提取物浓度呈线性相关。长时间孵育后,标记的醛进一步还原为醇,表明生物发光细菌中也存在长链醛还原酶。在生物发光细菌的生长和发育过程中对提取物中脂肪酸还原酶活性的测量表明,脂肪酸还原酶活性与荧光素酶共同被诱导,这表明这些酶受到协同调节,并直接表明脂肪酸还原酶参与细菌生物发光系统中醛的生物合成。