CONVERSION OF ALDEHYDE TO ACID IN BACTERIAL BIOLUMINESCENT REACTION
CONVERSION OF ALDEHYDE TO ACID IN BACTERIAL BIOLUMINESCENT REACTION
复制标题
DOI：
10.1021/bi00748a016
复制
发表时间：
1973-01-01
期刊：
影响因子：
2.9
通讯作者：
MEIGHEN, EA
中科院分区：
文献类型：
--
作者：
DUNN, DK;MICHALISZYN, GA;MEIGHEN, EA
D. K. Dunn，GA Michaliszyn，I. G. Bogacki和E. a. Meighen * 摘要：细菌荧光素酶在还原的黄素单核苷酸、氧和长链脂肪醛存在下催化光的发射。醛的作用以及它是否直接参与化学反应还不清楚。在本研究中，醛在发光系统中的功能已被研究使用[1 - 14 C]癸醛在耦合酶系统。用于生物发光反应的底物（醛和FM-NH2）通过马肝醇脱氢酶和FMN还原酶在反应混合物中产生。对反应产物进行定量分析，并且在FMNH 2，O2和长链脂肪族醛的存在下，细菌荧光素酶的分布催化490 nm处的光发射（Cordendum和Strehler，1953; Strehler等人，1954年）。虽然醛对于高量子产率是必不可少的，但这种分子在生物发光反应中的命运是未知的。许多工作人员的研究表明，发出的光的总量与添加的醛的量成比例（McElroy和绿色，1955; Cordycan和Totter，1957; Lee，1972; Shimomura等人，1972年）。有人提出，醛转化为相应的长链酸，因为该反应将提供发射490 nm量子光的能量（McElroy和绿色，1955）。最近对这一建议的支持来自于通过质谱法检测生物发光反应混合物中的酸（Shim-omura等人，1972年）。然而，重要的是要确定在不存在荧光素酶的情况下不发生酸产生。事实上，荧光素酶的缓慢周转（u/.=在本研究中，我们研究了醛在生物发光反应混合物和缺少发光必需组分的对照中的去向。在对照组和发光系统中观察并定量酸的产生。生物发光反应混合物和对照之间酸产生的净差异与发射光的总量直接相关。这一结果与最近的一项研究结果一致。
D. K. Dunn, GA Michaliszyn, I. G. Bogacki, andE. A. Meighen* abstract: Bacterial luciferase catalyzes the emission of light in the presence of reduced flavine mononucleotide, oxygen, and a long chain aliphatic aldehyde. The role of the aldehyde and whether it directly participates in the chemical reaction have not been clear. In the present study, the function of the aldehyde in the luminescent system has been investigated using [l-14C] decanal in a coupled enzyme system. The substrates for the bioluminescent reaction (aldehyde and FM-NH2) were generated in the reaction mixture by horse liver alcohol dehydrogenase and FMN reductase. The reaction products were quantitatively analyzed and the distribution of acterial luciferase catalyzes the emission of light at 490 nm in the presence of FMNH2, 02 and a long chain aliphatic aldehyde (Cormier and Strehler, 1953; Strehler et al., 1954). Although the aldehyde is essential for high quantum yields, the fate of this molecule in the bioluminescent reaction is unknown. Investigations by a number of workers have shown that the total amount of light emitted is proportional to the amount of added aldehyde (McElroy and Green, 1955; Cormier and Totter, 1957; Lee, 1972; Shimomura et al., 1972). It has been proposed that the aldehyde is converted to the corresponding long chain acid since this reactionwould provide sufficientenergy for the emission of a quantum of light at 490 nm(McElroy and Green, 1955). Recent support for this proposal has come from the detection by mass spec-troscopy of acid in the bioluminescent reaction mixture (Shim-omura et al., 1972). However, it is important to establish that acid production does not occur in the absence of luciferase. Indeed, the slow turnover of luciferase (u/.= 2 sec with decanal) and low yield of products have been the major reasons for the delay in identification of the chemical products of the reaction.In the present studies, we have investigated the fate of the aldehyde in the bioluminescent reaction mixture and in con-trols missing a component essential for light emission. Acid production was observed and quantitatedboth in the controls and in the light emitting system. A net difference in the acid production between the bioluminescent reactionmixture and the controls was directly relatedto the total amount of emit-ted light. This result is in agreement with a recent communica-