A Rapid, Scalable Method for Plant-Made Production of Cellulases

一种快速、可规模化的植物纤维素酶生产方法

• 批准号: 1067432
• 负责人: Karen McDonald
• 金额: $ 55万
• 依托单位: University of California-Davis
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1067432&HistoricalAwards=false
• 关键词: Rapid; Scalable; Method; Plant; Made

This NSF award by the Biotechnology, Biochemical and Biomass Engineering program supports the development of technologies and processes for efficient, rapid and scalable production of cellulase enzymes in plant tissues. To meet the requirements of the Energy Independence and Security Act of 2007, conversion of cellulosic biomass into advanced biofuels will require a significant amount of cellulose degrading enzymes at a low cost. Current methods for production of cellulose and hemicellulose degrading enzymes rely primarily on submerged fungal fermentation and are expensive, energy intensive, generate carbon dioxide, and consume cellulosic feedstock for growth of the production microorganisms. In this project, multiple thermostable cell wall degrading enzymes will be produced in Nicotiana benthamiana (a relative of tobacco) plants and harvested leaves, utilizing novel transient expression systems. Advantages of the transient approach over in-planta production of enzymes in stably transformed plants are that it is rapid, can utilize wild type (non-transgenic) plant biomass, and that the plant growth and enzyme production stages are decoupled and can be separately optimized.Intellectual merit: This research project will improve fundamental bioprocess engineering understanding of transient agroinfiltration processes, plant viral amplicon expression systems, recovery and characterization of plant-made enzymes, scale-up considerations in transient agroinfiltration processes, and techno-economic and environmental assessment of alternative bioprocessing strategies for production of cellulase enzymes.Broader impact: The benefits of the proposed research to society are 1) cost-effective, energyefficient and environmentally-sound technologies that will not only be useful for the production of cellulase and hemicellulase enzymes to meet the U.S biofuels mandates but also lead to "greener" and more efficient bioproduction technologies for the industrial enzyme industry, 2)enhanced fundamental process understanding of recombinant protein production using transient agroinfiltration in plants which will accelerate growth of companies utilizing similar technologies for vaccine and therapeutics production, 3) broadening participation of women and underrepresented minorities through proactive recruitment and interdisciplinary research and educational training opportunities at the interface of plant science, biotechnology and bioprocess engineering, and 4) broad curricular impact by incorporating research findings into existing and new lecture and laboratory courses.

这项由生物技术、生物化学和生物质工程项目颁发的 NSF 奖项支持开发在植物组织中高效、快速和可扩展地生产纤维素酶的技术和工艺。为了满足 2007 年《能源独立与安全法案》的要求，将纤维素生物质转化为先进的生物燃料将需要大量低成本的纤维素降解酶。目前生产纤维素和半纤维素降解酶的方法主要依赖于深层真菌发酵，并且昂贵、能源密集、产生二氧化碳并消耗纤维素原料用于生产微生物的生长。在该项目中，将利用新型瞬时表达系统在本塞姆氏烟草（烟草的近亲）植物和收获的叶子中生产多种热稳定性细胞壁降解酶。与稳定转化植物中的植物内酶生产相比，瞬时方法的优点是速度快，可以利用野生型（非转基因）植物生物量，并且植物生长和酶生产阶段是分离的并且可以单独优化.智力价值：该研究项目将提高对瞬时农杆菌渗透过程、植物病毒扩增子表达系统、植物酶的回收和表征、瞬时农杆菌渗透过程的放大考虑因素以及技术经济的基本生物工艺工程理解以及对生产纤维素酶的替代生物加工策略的环境评估。更广泛的影响：拟议研究对社会的好处是 1) 具有成本效益、节能和环保的技术，不仅可用于生产纤维素酶和半纤维素酶酶不仅可以满足美国生物燃料的要求，而且还可以为工业酶行业带来“更绿色”和更高效的生物生产技术，2）利用瞬时农杆菌渗透增强对重组蛋白生产的基本工艺了解植物将加速利用类似技术进行疫苗和治疗药物生产的公司的发展，3）通过在植物科学、生物技术和生物过程工程的交叉领域积极招聘和跨学科研究和教育培训机会，扩大妇女和代表性不足的少数群体的参与，以及4）通过将研究成果纳入现有和新的讲座和实验室课程，产生广泛的课程影响。