MECHANISTICALLY CRYPTIC ENZYME REACTIONS

机械上神秘的酶反应

• 批准号: 3289941
• 负责人: John M. Schwab
• 金额: $ 9.79万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3289941
• 关键词: Clostridium; cyclic compound; cyclohexanones; deuterium; ethylene oxide; nuclear magnetic resonance spectroscopy; radiotracer; stereochemistry; tritium

The study of two mechanistically cryptic enzyme-catalyzed processes is proposed as are syntheses of a variety of useful, chirally-labeled compounds. An enzyme system from Clostridium aminobutyricum dehydrates 4-hydroxybutanoyl-CoA to E-2-butenoyl-CoA. This process appears to involve the removal of a relatively nonacidic proton and is therefore mechanistically unusual. Proposed studies include determining the overall reaction stereochemistry, evaluating characteristics of an enzyme-catalyzed allylic rearrangement that may be involved, and observing the fates of fluorine-substituted substrates. Substrates and substrate analogs, substituted with deuterium and/or fluorine will be synthesized. 2H and 19F NMR spectroscopy will be used for analyses. Elucidation of the mechanism will help to define the structural and mechanistic requirements of enzyme-catalyzed dehydrations in general. Such information can be of great utility in predicting pathways of secondary metabolism as well as metabolism of foreign substances, including toxins and drugs. Enzyme-level characterization of the oxidation of cyclohexanone to adipic acid by the anaerobe Paracoccus sp. NCIB 11086 is also proposed. Reasonable but unprecedented mechanisms will be evaluated, using substrates labeled with 13C and 2H; analyses will be by mass spectrometry. Elucidation of a unique mechanism may facilitate targeting of drugs to anaerobic pathogens. Applications of chirally-labeled oxirane to the synthesis of a variety of useful, labeled biomolecules are proposed.

两种机制隐蔽的酶催化的研究 提出了合成各种有用的， 手性标记的化合物。 酶系统， 氨基丁酸梭菌使4-羟基丁酰辅酶A脱水 转化为E-2-丁烯酰-CoA。 这一过程似乎涉及 去除相对非酸性的质子，因此 不寻常的机械。 建议的研究包括确定 整体反应立体化学，评价特征 酶催化的烯丙基重排， 参与，并观察氟取代的命运 印刷受体. 底物和底物类似物，用 氘和/或氟将被合成。 2 H和19 F NMR 光谱学将用于分析。 阐明 机制将有助于确定结构和机制 酶催化脱水的一般要求。 等 信息在预测疾病的途径方面具有很大的实用性， 次级代谢以及外源代谢 物质，包括毒素和毒品。 酶水平 环己酮氧化制己二酸的表征 由厌氧菌Paracoccus sp. NCIB 11086也提出。 将对合理但前所未有的机制进行评估， 使用13 C和2 H标记的底物;分析将通过 质谱分析法来 阐明一种独特的机制， 促进药物靶向厌氧病原体。 手性标记的环氧乙烷在合成一种 提出了多种有用的标记的生物分子。