SBIR Phase I: Cyanobacteria Metabolic Engineering for Dye Stability

这项小企业创新研究（SBIR）第一阶段项目的广泛影响是生产一种替代食品、化妆品和纺织工业中使用的石油基色素。该项目开发的基因工程技术可以改变全球天然色素产业。这些行业的公司正在寻求使用天然染料来替代石油基染料，但一直无法找到合适的替代品来替代蓝色。今天使用的大多数颜色都是由石油和不可再生材料制成的，这些材料会造成污染。这个项目通过使用特殊的微生物来解决可持续生产蓝色染料的问题。通过微生物生产成功地设计出一种稳定的天然蓝色染料，食品、化妆品和纺织行业生产的几乎任何产品都将能够取代供应链上的人工和合成染料。这项技术为新的天然制造技术建立了一个新的可扩展的生产平台，用于生产超越天然颜色的高价值产品。拟议的项目解决了目前用于食品、化妆品和纺织品的合成、不可持续、不健康的石油基蓝色染料的问题。这是由于在正常的制造过程中，天然蓝色颜料在长时间暴露于高温和酸性环境后会降解和褪色。该项目的研究目标是利用遗传和筛选方法，使光合作用制造一种长时间耐酸和耐高温的蓝色染料。目标是表征由蓝藻菌生产的现有天然蓝色染料的稳定性和经济性，在另一种微生物中表达该天然蓝色染料的部分，通过定向进化，合理设计和控制工程方法设计这些部分的稳定性，通过迭代过程来测试生长和稳定性。最后，工程基因将通过一种新的基于crispr的基因编辑技术在原始蓝藻中表达，并且将测试新型天然蓝色的酸性和高温稳定性。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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 of this Small Business Innovation Research (SBIR) Phase I project is to produce a replacement for petroleum-based colors used in the food, cosmetic and textile industries. The genetic engineering techniques developed in this project can change the global natural color industry. Companies in these industries are seeking to use natural dyes as a replacement for petroleum-based dyes, but have been unable to find a suitable replacement for the color blue.Most colors used today are made from petroleum and non-renewable materials that can pollute. This project addresses the sustainable production of blue dye by using special microbes. By successfully engineering a stable natural blue dye through microbial production, virtually any product produced by the food, cosmetic and textile industry will be able to replace artificial and synthetic dyes in the supply chain. This technology establishes a novel scalable production platform for the new natural manufacturing techniques for high-value products beyond natural colors. The proposed project addresses the problem of synthetic, unsustainable, unhealthful petroleum-based blue dye that is currently used in food, cosmetic and textile products. This is due to how natural blue pigments degrade and fade in color after prolonged exposure to high temperature and acidic environments during normal manufacturing processes. This project’s research goal is to use genetic and screening methods to enable photosynthetic manufacture of a blue dye tolerant of acids and high temperatures for an extended period of time. The objectives are to characterize the stability and economics of an existing natural blue dye produced by cyanobacteria, express the parts of that natural blue dye in another microbe, engineer the stability of those parts via directed evolution, rational design, and a controlled engineering approach through an iterative process to test for growth and stability. Lastly, the engineered genes will be expressed in the original cyanobacterium via a novel CRISPR-based gene editing technique, and the novel natural blue color will be tested for acid and high-temperature stability.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.