SBIR Phase II: Engineering brewer's yeast for enhanced flavor production during fermentation

小型企业创新研究(SBIR)第二阶段项目的更广泛影响/商业潜力是开发工程啤酒酵母，在啤酒发酵过程中生产啤酒花风味化合物，以取代对农业生产啤酒花的需求。使用农业生产的啤酒花作为风味成分带来了几个挑战：1)啤酒花种植是自然资源密集型的，2)啤酒花所赋予的风味不同于每一批，3)漫长的农业时间线使需求与供应脱钩，以及4)新的风味品种受到缓慢和资本密集型啤酒花育种计划的限制。该项目建立了一个产品开发框架，用于改造啤酒酵母，使其在发酵过程中产生符合消费者偏好的风味化合物。在该项目期间开发的技术将允许可持续、一致、按需生产啤酒花风味，此外，还可能扩展到其他口味和其他可发酵产品。这个SBIR第二阶段项目将开发工程啤酒酵母，用于生产通常从啤酒花中提取的风味化合物。目前，啤酒业依靠农业生产的啤酒花来赋予啤酒丰富的感官风味和特征。工程酵母菌株将替代传统酿造菌株，因为它们都可以发酵啤酒，并在成品啤酒中产生所需浓度的风味化合物。通过将灵活的开发框架应用于酵母菌株的开发，将产生新的啤酒酵母菌株，产生啤酒酿造商和消费者喜欢的风味香气。编码产生风味化合物的生物合成途径的基因将被整合到酿酒酵母中，并将测试各种基因调控程序，这些程序将在成品啤酒中产生无数的风味气味。菌株将在小规模微好氧发酵中进行初步评估，并通过GC/MS分析以筛选相关风味化合物浓度。根据筛选结果，具有所需目标风味化合物浓度的菌株将用于工业中试生产啤酒，并由感官分析小组进行评估，作为产生消费者反馈的一种手段。这一工作流程将以迭代的方式进行部署，以优化啤酒花风味酿造者酵母的性能。该奖项反映了NSF的法定使命，并已通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop engineered brewer's yeast to produce hop flavor compounds during beer fermentation, so as to replace the need for agriculturally produced hops. Using agriculturally produced hops as a flavor component poses several challenges: 1) hops farming is natural resource intensive, 2) flavors imparted by hops are inconsistent from batch-to-batch, 3) lengthy agriculture timelines decouple demand from supply, and 4) new flavor varieties are restricted by slow and capital intensive hops breeding programs. This project establishes a product development framework for engineering brewer's yeast to produce flavor compounds during fermentation that are matched to consumer preferences. The technology developed during this project will allow for sustainable, consistent, on-demand production of hop flavors, and, in addition, may be extended to additional flavors and other fermentable products.This SBIR Phase II project will develop engineered brewer's yeast for production of flavor compounds that are ordinarily derived from hops. At present, the beer industry relies on agriculturally produced hops to impart organoleptically rich flavors and character to beer. Engineered yeast strains will serve as a drop-in replacement for conventional brewing strains, in that they both ferment beer and produce flavor compounds at concentrations desired in the finished beer. By applying an agile development framework towards yeast strain development, new brewer's yeast strains will be generated that produce flavor bouquets preferred by brewers and consumers. Genes that encode biosynthetic pathways for production of flavor compounds will be incorporated into brewer's yeast, and various gene regulatory programs will be tested that give rise to myriad flavor bouquets in finished beer. Strains will be initially evaluated in small-scale micro-aerobic fermentations and analyzed by GC/MS to screen for relevant flavor compound concentrations. Based on screening results, strains with desired target flavor compound concentrations will be used to produce beer at industrial pilot scale for evaluation by sensory analysis panels as a means of generating consumer feedback. This workflow will be deployed in an iterative fashion to optimize the performance of the hops flavor-producing brewer's yeast.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.