发酵草本能源植物产甲烷菌群的宏基因组与宏蛋白质组学研究

Biogas is one of the new-style renewable energy sources with the greatest potential for development and a promising prospect for exploitation. In this research, a biogas-producing microbial community NYZ was taken as the material, which was an efficiently fermented herbaceous energy plants obtained from the previous studies. In combination with the emerging metagenomics and metaproteomics technologies in recent years, this proposal studied the vital movement of the biogas-producing microbial community NYZ at the peak of gas-generating based on the gene expressions and functions of microorganisms. The 454 GS FLX sequencing platform was adopted to make metagenome sequencing and sequence analysis of NYZ, and metaprteomics was employed to analyze and identify the NYZ holoprotein, based on which the structure of NYZ at the peak of gas-generating and the cooperative relations among different kinds of microorganism was clarified, and the gene expressions, functions and metabolism of NYZ at the peak of gas-generating was also revealed. The results will enable us to know more about the structure and functions of the biogas-producing microbial community, and provide guidance on how to regulate and control the metabolic activities of the biogas-producing microbial community, improve the structure of the biogas-producing microbial community and enhance the capacity of methane generation, which will provide theoretical basis and technical support for preparing biogas by exploiting and utilizing herbaceous energy plants.

生物燃气是最有发展潜力的新型可再生能源之一，开发前景十分广阔。本课题以前期研究获得的高效发酵草本能源植物产甲烷菌群NYZ为材料，联合应用近年来新兴的宏基因组学和宏蛋白质组学技术，在微生物基因表达与功能水平上，研究产甲烷菌群NYZ产气高峰时期的生命活动。通过454 GS FLX测序平台进行菌群NYZ宏基因组测序及序列分析，通过宏蛋白组学对菌群NYZ全蛋白质进行分析鉴定，阐明产气高峰时期产甲烷菌群的结构及不同种类菌之间的相互协作关系，揭示产气高峰时期产甲烷菌群中微生物的基因表达、功能与代谢。研究结果将加深对产甲烷菌群的结构与功能的认识，为调控产甲烷菌群代谢活动、改进菌群结构及提高产甲烷能力提供指导，为开发利用草本能源植物制备生物燃气提供理论依据和技术支持。

本项目以优质的草本能源植物巨菌草为原料，来研究发酵草本能源植物产甲烷菌群NYZ的宏基因组与宏蛋白质组。获得了产甲烷菌群样本的宏基因组测序数据，构建了产气高峰期产甲烷菌群的宏基因组数据库， 得到163,232个平均长度为677 bp的ORFs（gene），其总长为110,612,865 bp。发现菌群有5个门，9个纲，10个目，22个科，28个属，36个种组成。菌群的宏蛋白质学分析，共鉴定蛋白质组数2157个，成功构建了发酵草本能源植物产甲烷菌群的宏蛋白质组数据库。对菌群宏蛋白质组学分析，原始数据中质谱定量到的蛋白质共计 1290 个。在此基础上，明确了菌群的结构及不同菌之间相互协作关系，并绘制了菌群甲烷代谢途径。在国内外核心期刊发表学术论文2篇，其中SCI收录2篇；申请发明专利5个，其中获得3个授权发明专利；培养博士生1名、硕士生3名。项目结果有助于揭示发酵草本能源植物产甲烷菌群的结构和功能，对沼气工程产业化发展具有一定促进作用。