STTR Phase I: A novel combinatorial technology for engineering product tolerance traits in yeast

STTR 第一阶段：一种用于工程酵母产品耐受性状的新型组合技术

• 批准号: 1321480
• 负责人: Helge Zieler
• 金额: $ 22.49万
• 依托单位: Primordial Genetics Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1321480&HistoricalAwards=false
• 关键词: STTR; Phase; novel; combinatorial; technology

This Small Business Technology Transfer (STTR) Phase I project proposes to engineer butanol tolerance in the yeast Saccharomyces cerevisiae using a novel combinatorial genetic technology designed to enable yield enhancements in microbial production organisms. Microbial production of renewable fuels and chemicals is rapidly expanding. However, toxic products or by-products and engrained metabolic fluxes often limit yields. Companies engaged in this space are actively searching for solutions. Novel genetic technologies are required that can help overcome these limitations. The proposed technology is designed as a new way to reprogram a cell and confer useful phenotypes. Synthetic genes are constructed using proprietary combinatorial strategies, limiting the ability of organisms to generate compensatory genetic or epigenetic changes. High-complexity expression libraries of these genes are transferred into the organism followed by selection or screening for desirable characteristics. The proposed experiments will demonstrate the feasibility of the technology in a screen for tolerance of butanol - a second-generation biofuel and important chemical precursor - using Saccharomyces cerevisiae, a yeast commonly used for alcohol production. The genes discovered during this work will be of direct interest to companies employing yeast for fuel and chemical production.The broader impact/commercial potential of this project, if successful, will be the enablement of improvements in microbes used for production of chemicals, fuels, pharmaceuticals, foods, and food ingredients. There is increasingly widespread use of microbial organisms (bacteria, fungi, yeasts, cyanobacteria, and algae) to produce these materials. Fuels and chemicals produced from such renewable sources generate roughly $75B in annual product sales, and this industry is experiencing rapid expansion. However, improving the yield and efficiency of production organisms is limited by the highly complex regulatory systems that govern yield, resistance, metabolism and growth. Current genetic methods are capable only of incremental improvements. Dramatic increases in yield and efficiency will require harnessing the massive combinatorial potential of genomes - the material on which natural evolution works. The proposed technology has multiple advantages over current approaches, especially regarding the probability of achieving a phenotype of interest, novelty of the active genes, transferability of the phenotype, speed and cost. This technology promises dramatically more effective creation of microbes with improved characteristics in multiple industries. It also will enable breakthroughs in our understanding of how important traits relating to growth, yield and productivity are encoded in biological systems.

这个小企业技术转让（STTR）第一阶段项目提出了使用一种新的组合遗传技术来设计酵母酿酒酵母中的丁醇耐受性，该技术旨在提高微生物生产生物体的产量。可再生燃料和化学品的微生物生产正在迅速扩大。然而，有毒产品或副产品和根深蒂固的代谢通量往往限制产量。 从事这一领域的公司正在积极寻找解决方案。需要新的基因技术来帮助克服这些限制。这项技术被设计为一种重新编程细胞并赋予有用表型的新方法。合成基因是使用专有的组合策略构建的，限制了生物体产生补偿性遗传或表观遗传变化的能力。将这些基因的高复杂性表达文库转移到生物体中，然后选择或筛选所需的特征。拟议的实验将证明该技术在筛选丁醇耐受性方面的可行性-丁醇是第二代生物燃料和重要的化学前体-使用酿酒酵母（一种常用于酒精生产的酵母）。在这项工作中发现的基因将对使用酵母生产燃料和化学品的公司产生直接影响。如果成功，该项目的更广泛影响/商业潜力将是用于生产化学品，燃料，药品，食品和食品配料的微生物的改进。越来越广泛地使用微生物（细菌、真菌、酵母、蓝细菌和藻类）来生产这些材料。 由这些可再生能源生产的燃料和化学品每年产生约750亿美元的产品销售额，该行业正在快速扩张。然而，提高生产生物的产量和效率受到高度复杂的调控系统的限制，这些系统控制产量、抗性、代谢和生长。目前的遗传学方法只能逐步改进。产量和效率的大幅提高需要利用基因组的巨大组合潜力--自然进化所依赖的物质。所提出的技术与当前方法相比具有多个优点，特别是关于实现感兴趣的表型的概率、活性基因的新奇、表型的可转移性、速度和成本。这项技术有望在多个行业中更有效地创造具有改进特性的微生物。它还将使我们对与生长，产量和生产力相关的重要性状如何在生物系统中编码的理解取得突破。