COLLABORATIVE RESEARCH: Exploiting microbial hyperthermophilicity to produce an industrial chemical

1264052/1264053凯利/亚当斯微生物代谢工程的发展在很大程度上是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知识和相关分子遗传工具的可用性。然而，随着代谢工程针对越来越多的创新生物处理目标，迫切需要扩大潜在微生物宿主的范围。因为极端微生物已经被赋予了能够适应工业过程的严酷条件的特性，所以为这类微生物开发代谢工程能力是有意义的。直到最近，这一方向都是不可信的，但新发展的分子遗传学工具和基因组学研究为极端细菌的代谢工程开辟了前景。因此，这一拟议的项目将证明对高温考古菌进行代谢工程的有效性，该微生物在100摄氏度时生长最佳。为了证明确实可以对这种微生物进行代谢工程，这项工作将建立在最近在依赖温度的生化处理方面取得的成功的基础上，现在将从纤维二糖和麦芽糖中生产一种特殊化学品丁二酸酯。编码转运蛋白和代谢酶的基因从不太嗜热的生物中鉴定出来，最适温度接近70摄氏度，将被导入到重组呋喃假单胞菌菌株中。基因调控和产品形成的新方法战略性地利用超高温性将被用来优化在70℃附近生物产品的产量和效率。提供代谢工程超高温宿主产生非生理性产品的范例将为基于热的生物加工机会铺平道路。更广泛的影响将来自于学生在分子生物化学、功能基因组学和生物分子工程方面的跨学科培训，以学习极端微生物生物学和生物技术。每个秋季学期将招募两名本科生研究人员，并随后在PI的实验室进行暑期研究体验(UGA的一名生物化学家和NCSU的一名化学工程师)。这些学生将在暑假期间在另一个合作实验室度过两周，以促进跨学科培训。NCSU和UGA这一项目和相关项目的所有学生都将参加暑期在NSCU和UGA轮流举办的年度延长的“嗜热者小组会议”。这将促进跨学科培训和合作，并提供一个交流研究想法和方法的机制。

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.