SBIR Phase I: Development of a novel indigo dye process using biosynthesized molecules

SBIR 第一阶段：利用生物合成分子开发新型靛蓝染料工艺

• 批准号: 1938583
• 负责人: Tammy Hsu
• 金额: $ 22.49万
• 依托单位: HUUE, Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1938583&HistoricalAwards=false
• 关键词: SBIR; Phase; Development; novel; indigo

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project focus on improving the indigo dyeing process. Indigo, a dye integral to denim production, is produced on an industrial scale of 65,000+ tons annually by chemical synthesis, and the molecule must be solubilized with a reducing agent before dyeing. Both processes involve hazardous chemicals like formaldehyde and hydrogen cyanide that are damaging to dye workers, production facilities, and the surrounding environment. Aniline, the petroleum byproduct required to create indigo, is toxic when ingested, inhaled, or in contact with skin and may be carcinogenic. Reducing agents are deadly for aquatic life when dumped as wastewater in the dyeing process or costly to clean up as part of wastewater treatment dyeing. Today the indigo market is approximately $500 M, with indigo selling just under $10/kg. The proposed project will develop a novel biological method to synthesize the reduced form of indigo directly.This Small Business Innovation Research (SBIR) Phase I project will create a method to use Escherichia coli to biosynthesize an indigo precursor called indican, a stable, water-soluble molecule. With the introduction of a b-glucosidase, the indican is hydrolyzed and is converted to the water-soluble reduced indigo form, thus circumventing both the indigo chemical synthesis as well as the harmful reducing agent step. This indican method has been shown to work as an effective indigo dye on cotton fabric and yarns. To improve the process, the indican titer must be improved from the host strain. Two parallel strategies are proposed in this Phase I project: the indican pathway enzymes will be improved by directed evolution, and the fermentation process will be elongated by feeding key nutrients that the cell depletes, thus prolonging the productive lifespan of the fermentation. Currently the cells are fed tryptophan as a pathway precursor; this Phase I project also proposes engineering the cell to produce its own tryptophan.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.

该小型企业创新研究 (SBIR) 第一阶段项目的更广泛影响/商业潜力侧重于改进靛蓝染色工艺。靛蓝是牛仔布生产中不可或缺的染料，通过化学合成以每年 65,000 吨以上的工业规模生产，染色前必须用还原剂溶解该分子。这两个过程都涉及甲醛和氰化氢等危险化学物质，对染料工人、生产设施和周围环境造成损害。苯胺是制造靛蓝所需的石油副产品，摄入、吸入或与皮肤接触时有毒，可能致癌。当还原剂在染色过程中作为废水倾倒时，或者作为废水处理染色的一部分进行清理时，还原剂对水生生物来说是致命的。如今，靛蓝市场价值约为 5 亿美元，靛蓝售价略低于 10 美元/公斤。拟议的项目将开发一种新颖的生物方法来直接合成还原形式的靛蓝。这个小型企业创新研究（SBIR）第一阶段项目将创建一种使用大肠杆菌生物合成称为 indican 的靛蓝前体的方法，这是一种稳定的水-可溶性分子。通过引入β-葡萄糖苷酶，靛蓝被水解并转化为水溶性还原靛蓝形式，从而避免了靛蓝化学合成以及有害的还原剂步骤。这种靛蓝方法已被证明可以作为棉织物和纱线上的有效靛蓝染料。为了改进该过程，必须提高宿主菌株的指示滴度。该第一阶段项目提出了两个并行策略：通过定向进化改进印度途径酶，通过补充细胞消耗的关键营养物质来延长发酵过程，从而延长发酵的生产寿命。目前，细胞被喂食色氨酸作为途径前体；该第一阶段项目还建议对细胞进行改造以产生自己的色氨酸。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。