SBIR Phase I: Engineering Hydrolytic Enzymes for Enhanced Sugar Recovery From Biomass

这个小企业创新研究第一阶段项目旨在应用Protabit？S最先进的计算蛋白质设计(CPD)平台来设计一套更耐热、更具成本效益的木质纤维素酶，将生物质转化为单糖。CPD是一项创新技术，可生产具有传统蛋白质工程方法无法找到的新颖或增强特性的酶，Protabit正在积极开发可用的最先进的商业级CPD平台。在第一阶段，Protabit和加州理工学院的梅奥实验室将稳定两种重要的商业纤维素酶，H.julorina Cel5A和Cel7A。第一阶段的目标是：(A)应用CPD来设计耐热Cel5A和Cel7A变异体文库，并使用高通量筛选来确定在高温下具有最佳纤维分解活性的变异体；(B)测量最活跃的耐热变异体在多大程度上减少经过处理的玉米秸秆的酶负荷和水解时间，并估计相应的成本节约；以及(C)鉴定最活跃的耐热变异体，以获得其他商业上重要的特性，如pH值分布和表达产量。在第二阶段，Protabit和Caltech将同样设计其他关键的木质纤维素酶，并针对玉米秸秆、柳枝草和其他生物质饲料的活性优化这一核心集合。这项研究的更广泛影响/商业潜力是：(A)降低将生物质转化为葡萄糖和其他单糖的成本，这些是快速增长的可再生燃料和化学品行业的主要原材料投入；(B)展示Protabit的多功能蛋白质工程平台技术，解决工业生物技术中具有商业意义的特定问题。通过促进先进的替代生物燃料、乙醇以及合成橡胶、塑料和其他石油衍生材料的前体的生物生产，这项研究可以帮助减少美国对外国石油的依赖，并刺激国内制造业、投资和创造就业机会。此外，从地球上含量最丰富的聚合物纤维素中获取糖，可以减少对作为生物燃料原料的可食用玉米的需求，从而遏制粮食与燃料的辩论；它还鼓励种植能够在不适合粮食生产的边际土地上生长的专用饲料作物。此外，基于CPD的蛋白质稳定方法使许多领域的新产品和新技术成为可能，包括工业酶、蛋白质材料、新型抗体类支架以及具有更长保质期或生物半衰期的治疗方法。通过这个项目，Protabit和加州理工学院将帮助美国在经济、能源和环境安全方面迈出重要一步。

This Small Business Innovation Research Phase I project aims to apply Protabit?s state-of-the-art computational protein design (CPD) platform to engineer a more thermostable and cost-effective set of lignocellulosic enzymes for converting biomass to simple sugars. CPD is an innovative technology that yields enzymes with novel or enhanced properties that cannot be found using traditional protein engineering methods, and Protabit is actively developing the most advanced, commercial-grade CPD platform available. In Phase I, Protabit and the Mayo Lab at the California Institute of Technology will stabilize two important commercial cellulases, H. jecorina Cel5A and Cel7A. The objectives of Phase I are: (a) to apply CPD to design libraries of thermostable Cel5A and Cel7A variants, and to use high-throughput screening to identify variants with optimal cellulolytic activity at elevated temperatures; (b) to measure the degree to which the most active thermostable variants reduce enzyme loading and hydrolysis time on pretreated corn stover, and to estimate the corresponding cost savings; and (c) to characterize the most active thermostable variants for other commercially important properties such as pH profile and expression yield. In Phase II, Protabit and Caltech will similarly engineer other key lignocellulosic enzymes and optimize this core set for activity on corn stover, switchgrass, and other biomass feedstocks.The broader impacts/commercial potential of this research are: (a) to reduce the costs of converting biomass into glucose and other simple sugars, which are a principal raw material input in the rapidly growing renewable fuels and chemicals industries, and (b) to demonstrate Protabit's versatile protein engineering platform technology on a specific problem of commercial significance in industrial biotechnology. By facilitating the bio-based production of advanced drop-in biofuels, ethanol, and precursors for synthetic rubber, plastics, and other petroleum-derived materials, this research can help reduce U.S. dependence on foreign oil and spur domestic manufacturing, investment, and job creation. In addition, sourcing sugars from cellulose, the most abundant polymer on the planet, can curb the food-versus-fuel debate by reducing demand for edible corn as a biofuels feedstock; it also encourages the farming of dedicated feedstock crops capable of growing on marginal lands unsuitable for food production. Furthermore, CPD-based protein stabilization methods enable new products and technologies in myriad areas, including industrial enzymes, protein materials, novel antibody-like scaffolds, and therapeutics with improved shelf-life or biological half-life. With this project, Protabit and Caltech will help the U.S. take a major step toward economic, energy, and environmental security.