Collaborative Research: Engineering of Recoverable Cellulosomes for Bioconversion

合作研究：用于生物转化的可回收纤维素体工程

• 批准号: 1604526
• 负责人: Sergiy Minko
• 金额: $ 20.7万
• 依托单位: University of Georgia Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1604526&HistoricalAwards=false
• 关键词: Collaborative; Research; Engineering; Recoverable; Cellulosomes

The sustainable, industrial-scale production of transportation biofuels from plant biomass rely on enzymes called cellulases that break down the cellulosic fraction of biomass into sugars that can be fermented into bioethanol. The major cost of this process is the cellulase, which in the current generation of cellulosic biofuel facilities, is discarded after use because it must be dissolved in water to break down cellulose to sugars, and cannot be easily recovered. Recycling the cellulase enzyme would reduce the cost of cellulosic biofuels production. The goal of this project is to develop a new way of encapsulating the cellulase enzyme in recoverable solid form so that it can be collected reused multiple times before disposal. The key innovation is to bind the cellulase enzyme to nanometer-sized capsule called a cellulosome, which preserves the activity of the enzyme for cellulose conversion, and has a magnetic core which facilitates its recovery from dilute water processing streams. The educational activities associated with this project include mentoring of undergraduate student projects coordinated through the Nurturing American Tribal Undergraduate Research and Education (NATURE) program at North Dakota State University.Microorganisms that naturally produce cellulase enzymes bind them into special structures on the cell surface called cellulosomes. Due to the close proximity of these enzymes within the cellulosomes, the bioconversion of cellulosic and hemicellulosic materials proceeds with a high velocity and efficiency. However, in the industrial production of celluloytic enzymes, the cellulosome is not present. This research will engineer recoverable cellulosomes through a biomimetic approach where celluloytic enzymes are encapsulated within a nanstructured capsule containing a magnetic core. This biomimetic cellulosome will contain a number of complimentary celluloytic enzymes confined within a polymeric environment, and will be engineered to mimic several key properties of natural cellulosomes, including diversity of hydrolytic activity, close proximity of complementary enzymes, and strong and selective binding of the biomimetic cellulosome to cellulosic substrates. The magnetic core of the biomimetic cellulosome will facilitate its recovery from liquid suspension using magnetic separation techniques. To explore the potential of the biomimetic cellulosome for biomass conversion processes, the research has four objectives. The first objective is to make a series of new biomimetic cellulosomes to enhance synergism of hydrolytic enzymes for bioconversion of cellulosic materials to sugars. The second objective is to quantify the effects of the biomimetic cellulosome structure and immobilized enzyme diversity on cellulose hydrolysis to gain a fundamental understanding of cellulase synergism in natural cellulosomes. The third objective is to gain a fundamental understanding of cellulosome interaction with cellulosic substrates and lignin, and the fourth objective is to augment the functionality of the biomimetic cellulosome for broader applications. The anticipated outcomes from these studies include a fundamental understanding of the synergism of enzymes in engineered cellulosomes, their efficacy for different forms of substrates, and their potential for recovery and reuse.

从植物生物质中可持续地工业规模生产运输生物燃料依赖于称为纤维素酶的酶，该酶将生物质的纤维素部分分解成可发酵成生物乙醇的糖。 该方法的主要成本是纤维素酶，在当前一代的纤维素生物燃料设施中，纤维素酶在使用后被丢弃，因为它必须溶解在水中以将纤维素分解成糖，并且不能容易地回收。 回收纤维素酶将降低纤维素生物燃料生产的成本。 本项目的目标是开发一种新的方法，将纤维素酶以可回收的固体形式封装，以便在处理之前可以多次收集和重复使用。关键的创新是将纤维素酶结合到称为纤维素酶体的纳米大小的胶囊上，该胶囊保留了纤维素酶转化的活性，并具有磁芯，便于从稀水处理流中回收。 与该项目相关的教育活动包括通过北达科他州州立大学的培养美国部落本科生研究和教育（NATURE）计划协调的本科生项目的指导。天然产生纤维素酶的微生物将它们结合到细胞表面称为纤维素体的特殊结构中。 由于这些酶在纤维素体内非常接近，纤维素和半纤维素材料的生物转化以高速度和效率进行。 然而，在纤维素分解酶的工业生产中，不存在纤维素酶体。 这项研究将通过仿生方法设计可回收的纤维素酶，其中纤维素酶被封装在含有磁芯的纳米结构胶囊中。 这种仿生多纤维素酶体将含有限制在聚合物环境中的许多互补的纤维素分解酶，并且将被工程化以模拟天然多纤维素酶体的几个关键性质，包括水解活性的多样性、互补酶的紧密接近以及仿生多纤维素酶体与纤维素底物的强和选择性结合。 仿生纤维素酶体的磁性核心将有助于使用磁性分离技术从液体悬浮液中回收它。 为了探索仿生纤维素酶体在生物质转化过程中的潜力，本研究有四个目标。第一个目标是制备一系列新的仿生纤维素体，以增强水解酶的协同作用，用于纤维素材料的生物转化为糖。 第二个目标是量化的仿生纤维素酶体结构和固定化酶的多样性对纤维素水解的影响，以获得在天然纤维素酶体的纤维素酶协同作用的基本理解。 第三个目标是获得纤维素酶体与纤维素底物和木质素相互作用的基本理解，第四个目标是增加仿生纤维素酶体的功能以用于更广泛的应用。 这些研究的预期结果包括对工程化纤维素体中酶的协同作用、它们对不同形式底物的功效以及它们回收和再利用的潜力的基本理解。