*** 9661474 Deits This Small Business Innovation Research Phase I project to be carried out by Enzyme Design, Inc. will generate proof-of-principle data for a new enabling technology in enzyme expression, termed EnzyMatrixTM expression. EnzyMatrixTM expression adapts the major host microorganism in use in the enzyme industry, Bacillus subtilis, to create a new platform for enzyme expression. The product of EnzyMatrixTM expression is a particulate form of the target heterologous enzyme. The particulate nature of the product facilitates enzyme recovery, affording the potential to greatly simplify downstream processing. Since downstream processing costs dominate bioproduct selling prices, these savings can substantially expand the market for enzyme applications in the pharmaceutical and fine chemical industries. The particles themselves are heat, shear and organic chemical resistant, offering a platform for catalysts for use in a variety of industrial processes. The research objectives of this project are to express two enzymes with significantly different structures and functions using EnzyMatrixTM expression and to evaluate the properties of the resulting expressed enzymes; increase the level of expression on a per-cell and a per-liter fermenter basis, using one of the expressed enzymes as a model; carry out strain improvement to improve yield and speed EnzyMatrixTM expression; and evaluate chemical and genetic methods for stabilizing EnzyMatrixTM enzymes. The proposed enabling technology, the EnzyMatrixTM expression, will make it possible for Enzyme Design to offer a biocatalyst with the key advantages of rapid development and scaleup, uniform product properties and economical production. Through this technology the nation will realize improved efficiencies of production including reduced energy costs during processing and high production specificity, leading to reduction of the waste stream burden. ***
