SBIR Phase II: High-Throughput Combinatorial Polymer Bioconjugates Synthesis and Application in Biocatalysis

这一小型企业创新研究(SBIR)项目的更广泛影响/商业潜力包括白色生物技术领域的进步，该领域利用酶来创造有价值的工业产品。作为生物分子，酶比传统化学物质更难操纵，往往需要更广泛的开发才能适应工业或制药制造。这个SBIR项目将展示如何使用称为聚合物的特殊合成材料来稳定酶的性能。酶具有精确、独特的结构和功能，这反过来又是它们在复杂化学反应中起催化作用的关键。另一方面，合成聚合物尽管结构不那么精确，但可以合理地设计来承受或响应化学、热或生物条件。酶和合成聚合物的协同融合产生了一种化学性质更好的先进酶，导致了化学品、生物燃料和药品等有价值产品的新制造工艺；这些工艺应该需要更少的能源，使用更少的危险试剂，产生更少的废物。这个SBIR第二阶段项目建议开发一种组合合成设备，可以高通量地筛选具有所需性质(例如，温度、pH或有机溶剂稳定性)的酶-聚合物偶联物。到目前为止，只有低通量的合成和表征方法被应用于酶-聚合物偶联物的制备，将开发限制在每种蛋白质只有几种类型的聚合物修饰上，并且依赖于随机猜测来选择被测试的变体。因此，为了充分受益于目前市场上可获得的各种聚合物，重要的是开发方法来规模化地鉴定性能最佳的酶-聚合物偶联物。这将通过高通量合成酶-聚合物结合物和高通量筛选所获得的性质相结合来实现。建议研究的目标应用集中在药物生物催化方面。高通量方法的应用不仅将导致更快的研发周期，还将加快我们在识别可通过聚合物修饰实现的蛋白质属性方面的基础知识的发展，从而建立这种工业应用的方法。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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 manipulate 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 special synthetic materials known as 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 an advanced enzyme with improved chemical properties, leading to new manufacturing processes for valuable products such as chemicals, biofuels, and pharmaceuticals; these processes should require less energy, utilize fewer hazardous reagents, and generate less waste. This SBIR Phase II 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, it is important to develop methods to scale 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 attained properties. The target application of the proposed research is focused on pharmaceutical 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 identifying protein properties that can be achieved through polymer modification, thereby establishing this method for industrial applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.