Alcohol and hydrogen metabolism of hyperthermophilic microorganisms

拟议研究的长期目标是表征新型热稳定醇脱氢酶和氢化酶，并揭示它们作为生产乙醇和氢作为生物燃料的关键酶的作用和潜在应用。 超嗜热菌具有在90°C及以上的温度下生长的显着特性，并且它们能够代谢碳水化合物，包括各种生物聚合物和一些废物，以产生乙醇和氢气。然而，有非常有限的知识，所有的酶和它们的代谢调节，以及代谢途径参与超嗜热生产的酒精和氢气。 该研究的目标之一是研究从碳水化合物（包括甘油，生物柴油生产的副产品）生产酒精和氢气的机制。 将在各种生长条件下研究乙醇和氢气生产的调节，并将使用二维凝胶电泳和生化分析以及分子探针研究参与代谢途径的关键酶的鉴定。 将使用包括酶分析和三维结构测定在内的技术对酶进行纯化和表征其催化和生物物理特性。 将使用超嗜热菌可用的分子技术来实现酒精和氢气生产的优化。 本研究中使用的微生物包括来自细菌领域的代表（海栖热袍菌，Tmax =90°C，和海栖热袍菌，Tmax =90°C）。petrophila，Tmax =88°C）和热球菌（Thermococcus guaymasensis，Tmax =90°C）; kodakarensis KOD1，Tmax =100°C）。 这项研究的成功完成将提供有关超嗜热醇和氢代谢的进一步信息，并阐明在100°C附近温度下的代谢途径。 此外，所获得的结果将提供关键信息的替代和更有效的系统，用于生产生物燃料的酒精和氢气的建设，以及新的信息之间的进化关系的超嗜热和嗜温酶。

The long-term goal of the proposed research is to characterize novel thermostable alcohol dehydrogenases and hydrogenases, and to reveal their roles and potential applications as key enzymes for the production of alcohol and hydrogen as biofuels. Hyperthermophiles have a remarkable property of growing at temperatures of 90°C and above, and they are capable of metabolizing carbohydrates including various biopolymers and some waste materials to produce ethanol and hydrogen. However, there is very limited knowledge about all the enzymes and their metabolic regulation as well as metabolic pathways involved in the hyperthermophilic production of alcohol and hydrogen. One of the objectives of the research is to investigate the mechanisms of alcohol and hydrogen production from carbohydrates including glycerol, a by-product of biodiesel manufacture. The regulation of the production of ethanol and hydrogen will be studied under various growth conditions, and the identification of key enzymes involved in the metabolic pathways will be investigated using two-dimensional gel electrophoresis and biochemical analysis as well as molecular probes. The enzymes will be purified and characterized with respect to their catalytic and biophysical properties using techniques including enzymatic analyses and three-dimensional structural determination. The optimization of the production of alcohol and hydrogen will be achieved using molecular techniques available for hyperthermophiles. The microorganisms to be used in this study include representatives from the domains of Bacteria (Thermotoga maritima, Tmax =90°C, and T. petrophila, Tmax =88°C) and Archaea (Thermococcus guaymasensis, Tmax =90°C, and T. kodakarensis KOD1, Tmax =100°C). Successful completion of this research will provide further information about the hyperthermophilic alcohol and hydrogen metabolism, and elucidate the metabolic pathways at temperatures near 100°C. Furthermore, the results obtained will provide key information in the construction of alternative and more efficient systems for the production of biofuels alcohol and hydrogen, as well as new information on the evolutionary relationships between hyperthermophilic and mesophilic enzymes.