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

SBIR 第一阶段：工程水解酶以增强从生物质中回收糖

• 批准号: 1215234
• 负责人: Barry Olafson
• 金额: $ 14.95万
• 依托单位: Protabit LLC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1215234&HistoricalAwards=false
• 关键词: SBIR; Phase; Engineering; Hydrolytic; Enzymes

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.

该小型企业创新研究第一阶段项目旨在应用 Protabit 最先进的计算蛋白质设计 (CPD) 平台来设计一套更热稳定且更具成本效益的木质纤维素酶，用于将生物质转化为单糖。 CPD 是一项创新技术，可产生具有传统蛋白质工程方法无法发现的新颖或增强特性的酶，Protabit 正在积极开发最先进的商业级 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：红褐肉座菌 Cel5A 和 Cel7A。第一阶段的目标是： (a) 应用 CPD 设计热稳定 Cel5A 和 Cel7A 变体文库，并使用高通量筛选来鉴定在高温下具有最佳纤维素分解活性的变体； (b) 测量最活跃的热稳定变体减少预处理玉米秸秆上酶负荷和水解时间的程度，并估计相应的成本节省； (c) 表征最活跃的热稳定变体的其他商业重要特性，例如 pH 谱和表达产量。在第二阶段，Protabit 和加州理工学院将类似地设计其他关键的木质纤维素酶，并优化这一核心组合，以实现玉米秸秆、柳枝稷和其他生物质原料的活性。这项研究的更广泛的影响/商业潜力是：(a) 降低成本将生物质转化为葡萄糖和其他单糖，这些糖是快速增长的可再生燃料和化学品行业的主要原材料输入，以及 (b) 展示 Protabit 针对特定问题的多功能蛋白质工程平台技术在工业生物技术中具有商业意义。 通过促进先进的直接生物燃料、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基生产，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、投资和就业创建。 此外，从地球上最丰富的聚合物纤维素中采购糖，可以减少对作为生物燃料原料的食用玉米的需求，从而遏制食品与燃料的争论。它还鼓励种植能够在不适合粮食生产的贫瘠土地上生长的专用原料作物。此外，基于 CPD 的蛋白质稳定方法可以在许多领域实现新产品和技术，包括工业酶、蛋白质材料、新型抗体样支架以及具有更长保质期或生物半衰期的疗法。 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。