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

这一小企业创新研究第一阶段项目的更广泛影响/商业潜力将是由不与食品来源竞争的可再生原材料制成的表面活性剂(酰乙醇胺)的商业化。表面活性剂是清洁产品中的泡沫成分，使其具有清洁能力。表面活性剂是由石油或棕榈油等种子油制成的。这些原材料的使用增加了温室气体污染，并助长了对热带雨林的砍伐。社会要求环境可持续(更绿色)的产品减少或去除有毒物质。对更环保化学品的需求创造了用更环保的替代品取代今天的表面活性剂的机会。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，因为没有使用有毒溶剂，而且表面活性剂是可生物降解的，由糖等可再生原材料生产，因此不会增加温室气体积累。酰基乙醇胺表面活性剂旨在取代目前在制造过程中被致癌物质1，4二氧六环污染的当前商业表面活性剂。本文所述方法生产的表面活性剂不含任何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。这一点意义重大，因为非离子表面活性剂约占300亿美元表面活性剂市场的40%。这项一期研究项目的技术目标是构建一株能够产生酰氨基醇表面活性物质的芽孢杆菌，即酰基乙醇胺。某些天然存在的多肽合成酶催化特定氨基酸与其他特定氨基酸的连接。此外，某些多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产生独特的分子，如甘氨酸酰酯(与甘氨酸相连的脂肪酸)。在酶促合成甘氨酸酰酯的过程中，甘氨酸通过硫代酯键共价连接到合成酶上。产物的释放由硫代酯结构域催化。硫代酯酶的释放会产生与氨基酸甘氨酸相连的脂肪酸。某些自然产生的多肽合成酶利用还原酶域来释放产物。我们假设，通过还原酶结构域释放酰基甘氨酸将导致合成酰基乙醇胺，而不是酰基甘氨酸。这一阶段项目的目标是创造一种嵌合型多肽合成酶，在酶释放产物的过程中将酰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.