ENZYMES OF THE META-FISSION PATHWAY

本研究的长期目标是彻底定义 每种酶在亚裂变中的精确化学机制 通路 对这些酶的研究具有相当大的智力 以及实际意义。 这些研究应该阐明 机制和立体化学在进化中的作用 分解代谢途径，因为它们适应特定的底物。 在 此外，芳香族化合物的生物降解代表了一个 解决有毒废物污染问题的潜在办法。 在此期间，我们将集中精力在三个关键领域。 酶c，f该途径：4-巴豆酸互变异构酶，4- 巴豆酸脱羧酶和乙烯基丙酮酸水合酶。 这些 酶代表了一种常见的 通过亚裂变降解化合物的化学策略 路线 这些探索性研究将作为基础， 随后的详细机制研究和初步 2-HOD水解酶分支的研究。 我们的具体目标，按优先顺序列出，在此期间， 期间将是： (1)拟定中间体的明确鉴别 亚裂变途径的4-异巴豆酸分支。 (2)酶的非酶化学的表征 4-异巴豆酸分支的中间体。 (3)4-异巴豆酸酯立体专一性的测定 互变异构酶 (4)乙烯基丙酮酸水合酶作用的测定： 水合酶或水解酶？ (5)测定了4-甲基-3-（4-甲基-4-（4-甲基-4-（4-甲基-4-氧代苯基）氨基）-3-甲基-3-（4-甲基-4-氧代苯基）氨基甲酸酯的立体化学进程。 异巴豆酸脱羧酶。 这些基础研究应该作为遗传学的基础。 操作微生物以满足特定解毒 需求 此外，亚裂变途径的酶出现在 作为构建新型降解剂的细菌模型 途径。 这些研究将为研究更多的 在其他核反应堆中发现的亚裂变途径的复杂变化 有机体 最后，这些研究将增加对以下方面的了解： 一些基本的和有趣的酶学过程。

The long term goal of this research is to thoroughly define the precise chemical mechanisms of each enzyme in the meta-fission pathway. Studies of these enzymes have considerable intellectual as well as practical implications. These studies should elucidate the role of mechanism and stereochemistry in the evolution of catabolic pathways as they adapt to specific substrates. In addition, biodegradation of aromatic compounds represent one potential solution to incumbent problem of toxic waste pollution. During this funding period we will focus our efforts on three key enzymes c,f this pathway: 4-oxalocrotonate tautornerase, 4- oxalocrotonate decarboxylase, and vinylpyruvate hydratase. These enzymes represent the essence of what appears to be a common chemical strategy used to degrade compounds by the meta-fission route. These exploratory studies will serve as a foundation for subsequent future detailed mechanistic studies and preliminary studies of the 2-HOD hydrolase branch. Our specific aims, listed in order of priority, during this funding period will be: (1) Unequivocal identification of the proposed intermediates of the 4-oxalocrotonate branch of the meta-fission pathway. (2) Characterization of the non-enzymatic chemistry of the intermediates of the 4-oxalocrotonate branch. (3) Determination of the stereospecificity of 4-oxalocrotonate tautomerase. (4) Determination of the role of vinylpyruvate hydratase: is it a hydratase or hydrolase? (5) Determination of the stereochemical course of the 4- oxalocrotonate decarboxylase. These basic studies should serve as a foundation for genetic manipulation of microorganisms to meet specific detoxification needs. Furthermore, the enzymes of the meta-fission pathway appear to be the model for bacterial construction of novel degradative pathways. These studies will set the stage for study of the more complex variations of the meta-fission pathway found in other organisms. Finally, these studies will increase knowledge about some basic and intriguing enzymological processes.