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

这个小型企业技术转移(STTR)第一阶段项目建议使用一种新的组合基因技术来设计酿酒酵母对丁醇的耐受性，旨在提高微生物生产有机体的产量。可再生燃料和化学品的微生物生产正在迅速扩大。然而，有毒产品或副产品以及根深蒂固的代谢通量往往会限制产量。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。这项拟议的技术被设计为一种重新编程细胞并授予有用表型的新方法。合成基因是使用专利组合策略构建的，限制了生物体产生补偿性遗传或表观遗传变化的能力。这些基因的高复杂性表达文库被转移到生物体中，然后选择或筛选所需的特征。拟议的实验将证明该技术在使用酿酒酵母筛选丁醇耐受性方面的可行性。丁醇是第二代生物燃料，也是重要的化学前体。酿酒酵母通常用于酒精生产。在这项工作中发现的基因将直接关系到使用酵母生产燃料和化学品的公司。如果该项目成功，其更广泛的影响/商业潜力将是使用于生产化学品、燃料、药品、食品和食品配料的微生物得到改进。越来越广泛地使用微生物(细菌、真菌、酵母菌、蓝藻和藻类)来生产这些材料。利用这种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，该行业正在经历快速扩张。然而，控制产量、抗性、新陈代谢和生长的高度复杂的调控系统限制了生产有机体的产量和效率的提高。目前的遗传方法只能进行渐进的改进。产量和效率的大幅提高将需要利用基因组的巨大组合潜力--自然进化所依据的材料。与现有方法相比，该技术具有多种优势，特别是在获得感兴趣的表型的可能性、活性基因的新颖性、表型的可转移性、速度和成本方面。这项技术承诺在多个行业更有效地创造具有改进特征的微生物。它还将使我们在理解与生长、产量和生产力相关的重要性状如何在生物系统中编码方面取得突破。

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.