纤维素酶系列增效蛋白的理性搜寻及其协同过程动力学研究

纤维素的致密结构和结晶化特性是制约纤维素酶水解的瓶颈问题。本项目在前期研究工作的基础上，根据自然进化机制，提出从产纤维素酶的系列微生物发酵液中系统寻找和分离具有协同纤维素酶水解作用的非酶蛋白，以期获得更多种类的新型纤维素酶增效蛋白。将选择有代表性的产纤维素酶的微生物进行发酵培养，提取、分离、纯化、鉴定获得目的非酶蛋白，并克隆其基因，建立非酶增效蛋白数据库。选择2-3种有代表性的新型增效蛋白研究其与纤维素酶协同水解过程，阐明其增效作用机理；通过检测降解过程中间物、产物变化及纤维原料的形态、结构及组成变化，分析不同外部条件对增效作用的影响，构建包括吸附、传质等作用的协同过程动力学模型，求取动力学参数，从而达到定性定量评估不同增效蛋白的增强作用。研究成果不仅可为构建纤维素水解基因工程菌提供有自主知识产权和实用价值的微生物及基因资源，也能成为纤维素水解过程优化和控制的科学依据。

The compact of the cellulose structure and the characteristics of crystallization are bottlenecks to the cellulose enzyme hydrolysis. Basedd on the previous work, the object of this project is to find for different kinds of novel cellulase-activity-enhancing proteins in cellulase-producing microorganism, which is proposed according to the hypothesis of natural evolution. The novel enhancing protein isolated from the typical cellulase-producing microorganism will be identified and cloned to construct the gene and protein database. Among the obtained typical novel cellulase-activity-enhancing proteins, 2-3 proteins will be studied as a model to clarify the synergetic mechanism between enhancing protein and cellulase by investigating the cellulose hydrolysis process. Through determining the medium and end products changes and the microstructure and morphological changes of substrates, the synergetic kinetic models will be constructed including absorption and mass transformation and the kinetic parameters calculated, so as to qualitatively and quantitatively evaluate the synergetic degree of different kinds of cellulase-activity-enhancing proteins.The expected research findings of this project may not only provide microorganism and gene resources with self intellectual property rights and practical value for construction of cellulose degrading gene engineering microorganism, but also provide the scientific basis for the optimization and regulation of the cellulose hydrolysis process.