SBIR Phase I: Production of an Acyl Ethanolamine Surfactant by Fermentation

这个小企业创新研究第一阶段项目的更广泛的影响/商业潜力将是由可再生原材料制成的表面活性剂（酰基乙醇胺）的商业化，这些原材料不会与食物来源竞争。表面活性剂是清洁产品的泡沫成分，赋予它们清洁能力。 表面活性剂由石油或种子油如棕榈油制成。 这些原材料的使用增加了温室气体污染，助长了雨林的砍伐。 社会需要环境可持续（绿色）的产品，减少或消除毒性。 对更环保化学品的需求为用更环保的替代品取代当今的表面活性剂创造了机会。该项目生产的表面活性剂化学品本质上比传统化学品更安全，因为不使用有毒溶剂，并且表面活性剂是可生物降解的，由糖等可再生原料生产，因此不会增加温室气体积累。 酰基乙醇胺表面活性剂旨在替代当前生产工艺中被致癌物1，4-二氧六环污染的当前市售表面活性剂。 通过本文所述的努力生产的表面活性剂将不含任何1，4二氧六环。 该项目的成功完成将展示一种生产非离子表面活性剂的新技术。 这是重要的，因为非离子表面活性剂占300亿美元表面活性剂市场的约40%。本一期研究项目的技术目标是构建一株能产生酰基氨基醇表面活性剂，即酰基乙醇胺的芽孢杆菌菌株。 某些天然存在的肽合成酶催化特定氨基酸与其他特定氨基酸的连接。 此外，某些肽合成酶催化特定脂肪酸与特定氨基酸的连接。 过去的工作表明，这个系统可以被改造成催化独特分子的产生，如酰基甘氨酸（脂肪酸连接到甘氨酸）。 在酰基甘氨酸的酶促合成期间，甘氨酸经由硫酯键共价连接至合成酶。 产物释放由硫酯结构域催化。 通过硫酯酶的释放导致与氨基酸甘氨酸连接的脂肪酸的产生。 某些天然存在的肽合成酶使用还原酶结构域来释放产物。 我们假设酰基甘氨酸通过还原酶结构域的释放将导致酰基乙醇胺的合成，而不是酰基甘氨酸。 该I期项目的目的是创建嵌合肽合成酶，其在从酶释放产物的过程中将酰基甘氨酸转化为酰基乙醇胺。

The broader impact/commercial potential of this Small Business Innovation Research Phase I project would be commercialization of a surfactant (acyl ethanolamine) made from renewable raw materials, which do not compete with food sources. Surfactants are the bubbly components of cleaning products that give them their cleansing power. Surfactants are manufactured from petroleum or from seed oils, such as palm oil. The use of those raw materials increases greenhouse gas pollution and contributes to deforestation of rainforests. Society is demanding environmentally sustainable (greener) products with reduction or removal of toxicity. The demand for greener chemicals creates an opportunity to replace today's surfactants with greener alternatives. The surfactant chemicals produced in this project are inherently safer than traditional chemicals because toxic solvents are not used, and the surfactants are biodegradable and produced from renewable raw materials such as sugars, and as a result do not contribute to increased greenhouse gas accumulation. The acyl ethanolamine surfactant is designed to replace current commercial surfactants, which are contaminated, during the current manufacturing process, with the carcinogen 1,4 dioxane. The surfactant produced by the effort described here will not contain any 1,4 dioxane. Successful completion of this project will demonstrate a new technology for the production of nonionic surfactants. This is significant since nonionic surfactants represent about 40% of the $30 billion surfactant market. The technical objective of this Phase I research project is to construct a Bacillus strain that produces an acyl amino alcohol surfactant, namely, acyl ethanolamine. Certain naturally existing peptide synthetase enzymes catalyze the linkage of particular amino acids to other particular amino acids. In addition, certain peptide synthetase enzymes catalyze the linkage of particular fatty acids to particular amino acids. Past work demonstrated that this system can be engineered to catalyze the creation of unique molecules, such as acyl glycinate (fatty acid linked to glycine). During enzymatic synthesis of acyl glycinate, glycine is covalently attached to the synthetase via a thioester bond. Product release is catalyzed by a thioesterse domain. Release by a thioesterase results in production of fatty acid linked to the amino acid glycine. Certain naturally occurring peptide synthetase enzymes use reductase domains to release products. We hypothesize that release of acyl glycine via a reductase domain will result in the synthesis of acyl ethanolamine, rather than acyl glycine. The objective of this Phase I project is to create a chimeric peptide synthetase enzyme that converts acyl glycine into acyl ethanolamine during the process of release of the product from the enzyme.