内蒙古原始森林土壤宏基因组文库构建及低温高活性纤维素酶筛选和特征分析

项目名称： 内蒙古原始森林土壤宏基因组文库构建及低温高活性纤维素酶筛选和特征分析

项目编号： No.31460002

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 孟建宇

作者单位： 内蒙古农业大学

项目金额： 48万元

中文摘要： 纤维素是解决石化能源短缺及其引起的环境污染问题的最具潜力的途径。现有的纤维素酶活性尚不够高，是现阶段纤维素无法替代石化能源的关键限制因素。而对于气温较低的地区，低温是纤维素利用的另一主要限制因素。研究筛选高酶活低温纤维素酶是解决问题的关键。当前普遍使用的、基于分离培养微生物而筛选高效低温纤维素酶并不成功。而宏基因组技术不依赖于培养，已获得了一些高活性的酶类。但基于宏基因组技术研究筛选低温纤维素酶尚未见研究报道。内蒙古原始森林较多，低温的气候特点突出，蕴藏有丰富多样的低温纤维素酶资源。本项目拟基于宏基因组技术，通过富集低温木质纤维素降解微生物及构建其宏基因组文库，然后以功能驱动方法筛选高酶活纤维素酶，并对其基因和预测蛋白结构进行分析，明确基因和蛋白的结构特点，阐明低温纤维素酶高效降解纤维素的分子机制，为充分认识和利用内蒙古地区低温纤维素酶资源提供理论依据和实践基础。

中文关键词： 宏基因组；纤维素酶；细菌；Fosmid；文库；功能驱动技术

英文摘要： Cellulose is the most potential solution of petrochemical energy shortage and environmental pollution problems. Existing cellulose enzyme activity is not high enough, which is the key limiting factor for cellulose cannot replace the fossil energy. For low temperature area, low temperature is another major limiting factor for cellulose using. Study of screening high enzyme activity of cold-adapted cellulase is the key to solve the problem. The current widely method used for screening of high efficiency cold-adapted cellulase based on the cultivation of microorganisms is not successful. While the metagenomics technology does not depend on the cultivation, and some highly active enzymes had been isolated by this technology. But the screening of cold-adapted cellulase based on the metagenomics technology has not been reported. There are many primeval forests with the climate characteristics of low temperature in Inner Mongolia. And there are rich varieties of cold-adapted cellulase resources in the primeval forests. This project is based on the metagenomics technology, and builds the metagenomic library through enrichment of lignocellulose-degrading microorganisms at low temperature. Screen high enzyme activity of cellulase by function driving assay method, analyze cellulase gene and predicted protein structure, and define the structure characteristics of specific genes and proteins to clarify the molecular mechanism of efficient degradation of cellulose by cellulase at low temperature. It will provide theoretical basis and practical basis for fully understanding and utilizing the cold-adapted cellulase resources in Inner Mongolia.

英文关键词： metagenomics;cellulase;bacteria;Fosmid library;function driving assay