SBIR Phase I: Alcohol Resistant Enzymes through High-Throughput Combinatorial Protein-Polymer Conjugate Synthesis

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project includes advancement of the field of white biotechnology, which utilizes enzymes to create valuable industrial products. As biological molecules, enzymes are more difficult to work with than conventional chemicals and often need more extensive development before they can be adapted to industrial or pharmaceutical manufacturing. This SBIR project will demonstrate how enzymes performance can be improved using stabilization with synthetic polymers. Enzymes are characterized by precise, unique structure and function, which is in turn essential for their role in catalysis of complex chemical reactions. Synthetic polymers, on the other hand, despite being less precisely structured, can be rationally designed to withstand or respond to chemical, thermal or biological conditions. The synergistic fusion of enzymes and synthetic polymers results in advanced nano-armored enzyme with improved properties such as solvent and temperature resistance, and modulated activity. Creating such novel stabilized enzymes will result in more efficient commercial utilization of enzymatic catalysis which requires less energy, utilizes less hazardous reagents, and generates less waste while generating valuable products such as chemicals, biofuels, and pharmaceuticals. This SBIR Phase I project proposes to develop a combinatorial synthesis device that can feed high-throughput screening of enzyme-polymer conjugates with desired properties (for instance, temperature, pH- or organic solvent stability). To date, only low-throughput synthesis and characterization methods have been applied to the preparation of enzyme-polymer conjugates, limiting development to only few types of polymer modification per protein and depending on stochastic guesswork to select the variants tested. Thus, in order to fully benefit from the diverse set of polymers currently available on the market one has to consider methods of scaling the identification of optimally performing enzyme-polymer conjugates. This will be achieved through combination of high-throughput synthesis of enzyme-polymer conjugates and high-throughput screening of gained properties. The initial target application of the proposed research is focused on the industrial biocatalysis. Application of a high-throughput method will not only result in faster research and development cycles, but also will accelerate our development of fundamental knowledge of what kind of protein properties can be gained through polymer modification, thereby establishing this method for industrial applications.