气藏相关的特异性甲烷氧化菌的拉曼光谱研究及其在天然气气藏现场、快速勘探中的应用

Methanotrophs act as the major indicator bacteria of gas pool, and its rapid, accurate discrimination plays a crucial role in developing novel microbial prospecting technologies for oil & gas. Highly sensitive and selective Raman spectroscopic technique hopefully provides a novel approach for methanotrophs discrimination without the interference of water. This project attempts to build up a Raman database based on Raman measurements of various species or phenotypes of methanotrophs and related biological molecules. Data mining and chemometrics methods will be utilized to process the mass data. Subsequently, data processing software and spectral identification module are developed according to the corresponding results of routine microbiological tests. The methane-oxidizing bacteria samples, which is located in near-surface soil on the top of gas reservoir, were chosen to deeply probe into the relationship between gas reservoir and characteristic Raman scattering properties of a certain number of specific methanotrophs. Further, the quantitative criteria and exploration module of microbial oil and gas exploration based on Raman analysis are established with the help of geological and seismic data. Combining the above mentioned modules and a home-built Raman fiber-optic probe will result in a portable gas reservoir prospecting instrument based on Raman studies of methane-oxidizing bacteria. This portable prospecting instrument could gradually be further applied in field exploration, which is expected to provide a new and practical technology for precise exploration of natural gas pools in China.

甲烷氧化菌是天然气藏勘探的主要指示菌，其快速精确检测是发展新型微生物勘探技术的核心和关键。高灵敏、高空间分辨、高光谱专一性和无需样品前处理的拉曼光谱技术可望为其提供一种全新研究手段。本项目从分子和细胞两个层次入手，检测与甲烷氧化菌相关的生物分子以及培养的不同种属与表型的甲烷氧化菌细胞的拉曼光谱，建立起生物分子和甲烷氧化菌的拉曼光谱数据库，并使用Data mining 技术和化学计量学手段对数据库进行解析与处理。研制出与常规微生物检验相关的数据处理软件和光谱鉴定模块。以气藏近地表土壤中甲烷氧化菌样品为研究对象，深入解析气藏属性与地表土壤甲烷氧化菌种属与含量的特征拉曼光谱的对应关系，进而在地质、地震等数据支撑下建立基于拉曼光谱技术的气藏微生物勘探的量化标准和勘探模块。将其与自制的拉曼光纤探针组装成便携式气藏微生物拉曼勘探仪并过渡到野外勘探，有望为我国天然气气藏的精确勘探提供一种新型实用技术。

本项目主要从分子和细胞两个层次入手，检测与甲烷氧化菌相关的生物分子以及培养的不同中暑与表型的甲烷氧化菌细胞的拉曼光谱和表面增强拉曼光谱，分别建立起生物分子和甲烷氧化菌的拉曼光谱和表面增强拉曼光谱数据库，并使用化学计量学手段对数据库进行解析和处理。研制出了与常规微生物检测相关的数据处理模型和光谱鉴别模块，并将其与自制的拉曼光谱光纤探测系统组装成专门现场、快速鉴别微生物细胞的便携式仪器；此外，完成了口腔黏膜组织与细胞、胃癌组织、胚胎培养基、血小板等生物组织和细胞自身拉曼散射性质的研究；更重要的是，通过目标物诱导来可控调节贵金属纳米粒子的聚焦状态，从而改变生物（模拟）体系的光学性质的变化（包括SPR或SERS信号等），藉此来实现一系列生物分子及金属离子等目标物的高选择性和高灵敏检测。其四，设计制备了一系列不同组成、形状和尺寸的贵金属纳米粒子及其复合纳米材料，以其作为SERS增强基底，直接检测有机小分子、生物大分子及微生物细胞。.此外，在国际交流与合作上，项目主持人胡继明教授先后出访德国、波兰、澳大利亚、加拿大、印度等国家开展国际合作和交流和参加学术会议，并作重要学术报告。在研究生培养上，共计培养博士研究生和硕士研究生近30名。受本项目资助，相关研究工作成果已有31篇SCI源刊论文发表在ACS Applied Materials & Interfaces (1), Scientific Reports (2)、Analytical Chemistry (2)、Chemical Communications (1)、Chemistry-A European Journal (1)、Journal of Materials Chemistry B (1)、Biosensors and Bioelectronics (2)等高影响因子刊物上。