Oxidation of Sulfur and Inorganic Sulfur Compounds in the Thermoacidophilic Archaeon Acidianus ambivalens

嗜热嗜酸古细菌 Acidianus ambivalens 中硫和无机硫化合物的氧化

• 批准号: 38211603
• 负责人: Privatdozent Dr. Arnulf Kletzin
• 金额: --
• 依托单位: Arbeitsgruppe Sulfur Biochemistry and Microbial Bioenergetics
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/38211603?language=en
• 关键词: Oxidation; Sulfur; Inorganic; Compounds; Thermoacidophilic

The thermophilic and acidophilic Archaeon Acidianus ambivalens oxidizes elemental sulfur to sulfuric acid as electron donor and energy source when growing aerobically. We purified several enzymes of the oxidation pathway recently including a soluble sulfur oxygenase reductase (SOR), a membrane-bound thiosulfate:quinone oxidoreductase (TQO) and a soluble tetrathionate hydrolase (TTH). In addition, we determined the 3D structure of the SOR. The goal of this proposal is to elucidate the reaction mechanisms of each of the three enzymes in molecular detail. With the SOR we plan to use a combination of site-directed mutagenesis, spectroscopy, rapid reaction kinetics of reaction intermediates, and X-ray crystallography of mutant proteins. With the TQO we want to establish gene expression, to do a systematic mutagenesis survey, and try to solve the 3D structure. With the TTH we want to identify the gene, establish gene expression and do a thorough biochemical characterization. With all three enzymes we will try to identify yet unknown inhibitors as probes for the active site structure.

嗜热嗜酸古生菌在需氧生长时，作为电子供体和能源，将元素硫氧化为硫酸。最近我们纯化了几种氧化途径的酶，包括可溶性硫氧合酶还原酶(SOR)、膜结合的硫代硫酸盐：苯醌氧化还原酶(TQO)和可溶性四硫酸水解酶(TTH)。此外，我们还确定了SOR的三维结构。这项建议的目的是从分子上详细阐明这三种酶的反应机理。对于SOR，我们计划使用定点突变、光谱、反应中间产物的快速反应动力学和突变蛋白质的X射线结晶学的组合。通过TQO，我们想要建立基因表达，进行系统的诱变调查，并试图解决3D结构。有了TTH，我们想要鉴定基因，建立基因表达，并做彻底的生化表征。对于这三种酶，我们将试图确定未知的抑制剂作为活性部位结构的探针。