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.
