Imaging Cellulose - Carbohydrate Binding Module Interactions With Nanometer Resolution Using Single Molecule Flourescence Methods

使用单分子荧光方法以纳米分辨率成像纤维素-碳水化合物结合模块相互作用

• 批准号: 1206908
• 负责人: Steve Smith
• 金额: $ 29.09万
• 依托单位: South Dakota School of Mines and Technology
• 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: 2012-08-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1206908&HistoricalAwards=false
• 关键词: Imaging; Cellulose; Carbohydrate; Binding; Module

ID: MPS/DMR/BMAT(7623) 1206908 PI: Smith, Steve ORG: South Dakota School of Mines and TechnologyTitle: Imaging Cellulose - Carbohydrate Binding Module Interactions with Nanometer Resolution Using Single Molecule Fluorescence MethodsINTELLECTUAL MERIT: Cellulose, one of the most abundant biopolymers on earth, is a naturally occurring nanomaterial. It is the primary component of plant cell walls and the feedstock for valuable products derived from biomass sugars, such as liquid and gaseous fuels. Most of what is known about the chemical structure of biomass comes from analysis of the products generated during its decomposition via batch methods. These approaches necessarily lose the spatial correlations between the biomolecules that drive biomass degradation and the chemical-spatial structure of the feedstock, that is, the plant cell wall and its primary constituents, cellulose, hemi-cellulose, and lignin. This project will focus on exploiting single molecule, super-resolution imaging methods to reveal important facets of cellulose-carbohydrate binding module (CBM) interactions that are lost in the ensemble averaging of batch analyses. CBMs play a critical role in the action of a large family of cellulase enzymes, locating and positioning the catalytic components on their target substrates. This project will utilize single molecule fluorescence methods to reveal molecular level details of the interaction of families of CBMs, derived from microbial, bacterial, and cellulosomal sources, with cellulose substrates. The binding characteristics of single CBMs, including specificity, orientation, motility, and binding persistence will be studied using CBMs tagged with fluorescent proteins (FPs), including photo-activated FPs, to extract orientation and positional information at the single molecule level. Sequential imaging of sparse subsets of single molecules with high resolution imaging approaching the 5nm level will be used to study these processes and the spatio-chemical structure of biomass.BROADER IMPACTS: This project aims to provide detailed knowledge of cellulose-carbohydrate binding module interactions, thus building a biophysical knowledge of the molecular processes responsible for the breakdown of cellulose to simple sugars, knowledge critical to enabling new biotechnologies, which could accelerate conventional bioprocessing. Such knowledge could, for instance, aid in selecting new enzymes or designing enzymatic bioprocessing systems. This work thus leverages the significant efforts made by the State of South Dakota towards the utilization of biomass in the production of biofuels and other biomass-derived products, and investments made toward stimulating this industry. The proposed work will enrich graduate and undergraduate education in South Dakota by enabling collaborative research opportunities and interdisciplinary training for scientists and engineers in the region. SDSM&T also maintains a permanent Native American outreach program, with established and well-organized outreach programs such as SD GEAR UP, aimed at increasing the number of Native American high-school students continuing on to college. On-going aspects of this project will be incorporated into the educational portions of this program.

ID: MPS/DMR/BMAT(7623) 1206908 PI：史密斯，史蒂夫ORG：南达科他州矿产与技术学院标题：成像纤维素-碳水化合物结合模块相互作用与纳米分辨率使用单分子荧光方法智力优点：纤维素，地球上最丰富的生物聚合物之一，是一种天然存在的纳米材料。它是植物细胞壁的主要成分，也是生物质糖衍生的有价值产品（如液体和气体燃料）的原料。对生物质化学结构的大部分了解来自于通过批处理方法对其分解过程中产生的产物的分析。这些方法必然会失去驱动生物质降解的生物分子与原料的化学空间结构之间的空间相关性，即植物细胞壁及其主要成分，纤维素，半纤维素和木质素。该项目将专注于利用单分子、超分辨率成像方法来揭示纤维素-碳水化合物结合模块（CBM）相互作用的重要方面，这些方面在批量分析的总体平均中丢失了。CBMs在一大家族纤维素酶的作用中起着关键作用，在其靶底物上定位和定位催化成分。该项目将利用单分子荧光方法揭示来自微生物、细菌和纤维素体来源的CBMs家族与纤维素底物相互作用的分子水平细节。利用荧光蛋白（FPs）标记的CBMs，包括光激活的FPs，在单分子水平上提取取向和位置信息，研究单个CBMs的结合特性，包括特异性、取向、运动性和结合持久性。接近5nm水平的高分辨率单分子稀疏子集序列成像将用于研究这些过程和生物量的空间化学结构。更广泛的影响：该项目旨在提供纤维素-碳水化合物结合模块相互作用的详细知识，从而建立纤维素分解为单糖的分子过程的生物物理知识，这些知识对于实现新的生物技术至关重要，可以加速传统的生物加工。例如，这些知识可以帮助选择新的酶或设计酶生物处理系统。因此，这项工作利用了南达科他州在利用生物质生产生物燃料和其他生物质衍生产品方面所做的重大努力，以及为刺激该行业而进行的投资。拟议的工作将通过为该地区的科学家和工程师提供合作研究机会和跨学科培训，丰富南达科他州的研究生和本科生教育。SDSM&amp；T还维持着一个永久性的印第安人外展项目，其中建立了组织良好的外展项目，如SD GEAR UP，旨在增加印第安人高中生继续上大学的人数。这个项目正在进行的方面将被纳入该计划的教育部分。