Functional genomics of the Clostridium thermocellum cellulosome

热纤梭菌纤维素体的功能基因组学

• 批准号: 5374166
• 负责人: Dr. Wolfgang H. Schwarz
• 金额: --
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5374166?language=en
• 关键词: Functional; genomics; Clostridium; thermocellum; cellulosome

The cellulosome of the thermophilic bacterium Clostridium thermocellum is a large, extracellular multienzyme complex consisting of more than 40 different modular enzymes, arranged on a non-catalytic structural protein (scaffoldin). The complex structure of the cellulosome is the basis for its extreme efficiency in the hydrolysis of crystalline cellulose, probably due to local synergism. Therefore the cellulosome is of the highest interest for the conversion of biomass. The genome of C. thermocellum has been sequenced, making functional genomics feasable. This project plans 1) to determine and to identify all components of the cellulosome with proteomic methods. 2) New components will be cloned with reverse genetics, sequenced and expressed for biochemical characterisation. 3) The composition of the whole complex will be investigated in vivo: changes in protein pattern after growth on different substrates point to the cellulosomal components necessary for the hydrolysis of crystalline cellulose. 4) Essential components will also be identified by their absence in induced non-cellulolytic mutants. 5) Analysis of the cellulosome-components bound to the bacterial cell wall gives an indication for the mechanism of attachment and assembly.

嗜热细菌Clostridium thermocellum的多纤维素酶体是一种大型的细胞外多酶复合物，由40多种不同的模块酶组成，排列在非催化结构蛋白（支架蛋白）上。纤维素酶体的复杂结构是其在结晶纤维素水解中的极高效率的基础，可能是由于局部协同作用。因此，纤维素酶体对于生物质的转化具有最高的兴趣。C.热纤蛋白已经测序，使功能基因组学变得非常困难。本计画计划1）以蛋白质组学方法，测定并鉴定多纤维素体的所有组成成分。2)新组分将通过反向遗传学进行克隆、测序和表达以进行生化表征。3)将在体内研究整个复合物的组成：在不同基质上生长后蛋白质模式的变化指向结晶纤维素水解所需的cellulosomal组分。4)必需组分也将通过其在诱导的非纤维素分解突变体中的缺失来鉴定。5)结合到细菌细胞壁的纤维素体组分的分析给出了附着和组装机制的指示。