Microbial Physiology of Alcohol and Hydrogen Production at High Temperatures

* 我的研究计划侧重于开发可再生能源，通过研究在高温下从大量可用的非食品生物质中高效生产酒精和H2的机制。这是通过表征和修饰参与代谢途径的新型酶和电子转移活性以及分离能够产生更大量醇或H2的新的超嗜热菌（在高温下茁壮成长的微生物）来实现的。并非所有参与酒精生产代谢途径的关键酶都是已知的，电子转移酶在提高高温下酒精和H2生产效率方面的调节作用也是未知的。这种缺乏了解阻碍了更有效的生物转化系统的发展。我的研究目标是：1）研究在90 ℃及以上生长的产醇和产氢超嗜热微生物的生理学和应用，2）表征它们的新型热稳定酶，如醇脱氢酶，脱氢酶，电子转移酶和氢化酶，并阐明了它们的代谢功能及其在生物催化和生产醇和H2作为生物燃料中的潜在应用。这将包括表征新的分离物，确定所研究的每种酶的生物物理和生化特性，研究它们在代谢途径中的催化作用，并探索蛋白质热稳定性的机制。生物技术的潜在应用包括绿色化学和大量可获得的非食品木质纤维素废料的更稳健的生物转化。将使用超嗜热古菌（丁基高温菌，Tmax=112 ℃，丁醇生产者）和细菌（海栖热袍菌，Tmax=90 ℃，乙醇和H2生产者）来研究与醇和H2代谢相关的机制。酶的表达和调节将使用生物化学和分子技术进行研究，关键酶纯化和表征。将相应地进行基因克隆和诱变。在发现的新分离株中，将对一两种进行研究。成功完成这项工作在加拿大将是独一无二的。它将提供进一步的见解超嗜热醇和H2代谢和蛋白质的热稳定性，并阐明在高温下生产醇和H2的代谢途径的调节。结果可以应用于构建更有效的绿色能源生产系统（例如，乙醇、丁醇、H2），并将用于对抗由消耗有限的化石燃料引起的气候变化和环境恶化的负面影响。HQP将获得进行微生物，分子生物学和生物化学研究的技能和经验，为他们在学术界和食品，生物技术和制药行业的职位做好准备。

***My research program focuses on developing renewable energy sources by studying the mechanisms of highly efficient alcohol and H2 production from largely available non-food biomass at high temperatures. This is achieved by characterizing and modifying novel enzymes and electron transfer activities involved in the metabolic pathways and isolating new hyperthermophiles (microbes that thrive at high temperatures) capable of producing greater amounts of alcohols or H2. Not all key enzymes involved in the metabolic pathways of alcohol production are known, nor are the regulatory roles of electron transfer enzymes in enhancing the efficiency of alcohol and H2 production at high temperatures. This lack of understanding hinders the development of more efficient bioconversion systems. My research objectives are to 1) investigate the physiology and applications of alcohol- and H2-producing hyperthermophilic microorganisms growing at 90 C and above, and 2) characterize their novel thermostable enzymes, such as alcohol dehydrogenases, aldehyde-producing enzymes, electron transfer enzymes, and hydrogenases, and elucidate their metabolic functions and potential applications in bio-catalysis and the production of alcohols and H2 as biofuels. This will include characterizing new isolates, determining the biophysical and biochemical properties of each enzyme studied, investigating their catalytic roles in metabolic pathways, and exploring mechanisms of protein thermostability. Potential applications in biotechnology include green chemistry and more robust bioconversion of largely available non-food lignocellulosic waste materials. The mechanisms related to alcohol and H2 metabolism will be investigated using both hyperthermophilic archaea (Hyperthermus butylicus, Tmax=112 C, a butanol producer) and bacteria (Thermotoga maritima, Tmax=90 C, an ethanol and H2 producer). The expression and regulation of enzymes will be studied using biochemical and molecular techniques, with key enzymes purified and characterized. Gene cloning and mutagenesis will be carried out accordingly. Of the new isolates found, one or two will be studied. Successful completion of this work will be unique in Canada. It will provide further insights into hyperthermophilic alcohol and H2 metabolism and protein thermostability, and elucidate the regulation of metabolic pathways for the production of alcohols and H2 at high temperatures. The results can be applied to the construction of more efficient systems for green energy production (e.g., ethanol, butanol, H2) and will serve to combat the negative impacts of climate change and environmental deterioration caused by consuming finite fossil fuels. HQP will gain skills and experience conducting microbiological, molecular biological and biochemical research, preparing them for positions in academia and the food, biotechnological, and pharmaceutical industries.