COLLABORATIVE RESEARCH: Exploiting microbial hyperthermophilicity to produce an industrial chemical

Kelly/Adams微生物代谢工程的发展主要基于有限数量的重组宿主（例如，大肠杆菌，酿酒酵母），因为这些微生物的生理学的大量知识和相关的分子遗传工具的可用性。然而，随着代谢工程越来越多地指向创新的生物加工目标，迫切需要扩大潜在微生物宿主的范围。由于极端微生物已经被赋予了能够适应工业过程中苛刻条件的特性，因此开发这类微生物的代谢工程能力是有意义的。直到最近，这一方向才变得难以置信，但新开发的分子遗传工具和基因组学研究为极端微生物的代谢工程开辟了前景。因此，这个拟议的项目将证明对极端嗜热古菌Pyrococcus furiosus进行代谢工程的有效性，该古菌在100°C下生长最佳。为了证明这种微生物确实可以进行代谢工程，这项工作将建立在最近在温度依赖性生化处理方面的成功基础上，现在将从纤维二糖和麦芽糖中生产一种特殊的化学物质琥珀酸盐。从最适温度值接近70°C的较不嗜热生物体中鉴定的编码转运蛋白和代谢酶的基因将被导入重组的P. furiosus菌株中。将利用战略性利用超嗜热性的基因调控和产物形成的新方法来优化70°C附近生物产物形成的产量和效率。提供一个范例的代谢工程一个极端嗜热宿主生产一个非生理产品将铺平道路，热为基础的，生物加工的机会。更广泛的影响将导致从跨学科的学生在分子生物化学，功能基因组学和生物分子工程研究极端微生物生物学和生物技术的培训。两名本科研究人员将在每个秋季学期招募，并随后在PI的实验室（一个在UGA和一个在NCSU化学工程师生物化学家）夏季研究经验的支持。这些学生将在夏季在另一个合作实验室度过两周，以促进跨学科培训。在NCSU和UGA这个和相关项目的所有学生将参加在夏季期间每年延长的“超嗜热菌小组会议”将在NSCU和UGA轮流主办。这将促进跨学科的培训和协作，并为交流研究思想和方法提供一个机制。

1264052/1264053 Kelly/AdamsThe development of microbial metabolic engineering has largely been based on a limited number of recombinant hosts (e.g., Escherichia coli, Saccharomyces cerevisiae) because of the substantial knowledge of the physiology of these microorganisms and availabilityof associated molecular genetic tools. However, as metabolic engineering is directed at more and more innovative bioprocessing objectives, there is a pressing need to expand the range of potential microbial hosts. Because extremophiles are already endowed with properties that are amenable to the harsh conditions characteristic of industrial processes, it makes sense to develop metabolic engineering capabilities for this group of microorganisms. This direction would have been implausible until very recently, but newly developed molecular genetic tools and genomics studies have opened up the prospects for metabolic engineering of extremophiles. As such, this proposed project will demonstrate the efficacy of metabolically engineering the hyperthermophilic archaeon, Pyrococcus furiosus, which grows optimally at 100°C. To demonstrate that this microorganism can, indeed, be metabolically engineered, the work will build upon recent success in temperature-dependent biochemical processing and will now produce a specialty chemical, succinate, from cellobiose and maltose. Genes encoding transporters and metabolic enzymes identified from less thermophilic organisms with optimum temperature values near 70°C will be imported into a recombinant P. furiosus strain. Novel approaches for gene regulation and product formation that strategically exploit hyperthermophilicity will be utilized to optimize yields and efficiencies for bioproduct formation near 70°C. Providing a paradigm for metabolically engineering a hyperthermophilic host to produce a non-physiological product will pave the way for thermally-based, bioprocessing opportunities.Broader impacts will result from the interdisciplinary training of students in molecular biochemistry, functional genomics and biomolecular engineering for studying extremophile biology and biotechnology. Two undergraduate researchers will be recruited during each fall semester and be supported subsequently for a summer research experience in the PI's laboratories (one biochemist at UGA and one chemical engineer at NCSU). These students will spend two weeks during the summer at the other collaborating lab to foster interdisciplinary training. All students on this and related projects at NCSU and UGA will participate in an annual extended 'Hyperthermophile Group Meeting' during the summer to be hosted at NSCU and UGA on an alternating basis. This will foster interdisciplinary training and collegiality, as well as provide a mechanism for exchange of research ideas and methodologies.