IN VIVO & IN VITRO NMR STUDIES OF METHANOGEN METABOLISM

体内

• 批准号: 3283214
• 负责人: Mary Fedarko Roberts
• 金额: $ 5.44万
• 依托单位: BOSTON COLLEGE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3283214
• 关键词: Methanobacteriaceae; acetyl coA; adenosine triphosphate; bioenergetics; carbon dioxide fixation; chromatography; electron transport; gluconeogenesis; methane; microorganism growth; microorganism metabolism; nitrogen metabolism; nuclear magnetic resonance spectroscopy; phosphoglycerate

Methanogens are fastidious anaerobes whose metabolism centers around the reduction of CO2 to CH4. These unique organisms have been classified a "archaebacteria" with characteristics evolutionarily distinct from procaryotes and eukaryotes. Many aspects of cellular metabolism in these organisms are unclear. In this proposal in vivo and in vitro NMR spectroscopy will be used to unravel carbon and nitrogen assimilation and specific aspects of bioenergetics in Methanobacterium thermoautotrophicum strain DeltaH. Specifically, we will (i) elucidate the exact role of 2,3-Cyclopyrophospho-glycerate (CPP), the major soluble carbon and phosphorus pool, in gluconeogenesis using 13C NMR; (ii) purify and characterize the enzymes that synthesize and degrade CPP; (iii) examine the unique CO2 fixation pathway that gives rise to asymmetric labeling of acetyl coenzyme A (using 13C NMR); (iv) further characterize nitrogen assimilation using 15N NMR; and (v) monitor cell bioenergetics with 31p NMR. The results of this work should lead to a better understanding of the metabolic role of CPP, (a novel intermediate in carbohydrate synthesis) and of how carbon and nitrogen assimilation are regulated in this methanogen, and to a comparison of metabolic pathways with other more common organisms.

产甲烷菌是一种要求严格的厌氧微生物，其代谢中心围绕 将CO2还原为CH 4。 这些独特的生物体被归类为 “古细菌”的特征在进化上不同于 原核生物和真核生物。 这些细胞代谢的许多方面 生物体不清楚。 在这个提议中，体内和体外NMR 光谱学将被用来解开碳和氮同化， 热自养甲烷杆菌中生物能量学的特殊方面 菌株Δ H。 具体而言，我们将（i）阐明 2，3-环焦磷酸甘油酸酯（CPP），主要的可溶性碳， 磷池，在异源生成中使用13 C NMR;（ii）纯化和 表征合成和降解CPP的酶;（iii）检查 独特的CO2固定途径，引起不对称标记， 乙酰辅酶A（使用13 C NMR）;（iv）进一步表征氮 使用15 N NMR进行同化;以及（v）用31 P监测细胞生物能量学 匪r 这项工作的结果应有助于更好地了解 CPP的代谢作用（碳水化合物合成的新型中间体）， 碳和氮的同化是如何在产甲烷菌中调节的， 以及与其他更常见的生物体的代谢途径的比较。