青海油田原油与采出液水相微生物差异机制及原油来源菌种资源多样性研究

Considering the growing demand for energy and the depletion of petroleum reservoirs, microbially enhanced oil recovery (MEOR) as low energy consumption, low environmental impact, and cost-effectiveness technique had attracted too many researchers. However, two rate limiting components of the MEOR development were present: ignorance the different microorganisms between crude oil and aqueous phases in petroleum reserves and lack of isolates from crude oils. Based on our previous study that significantly difference of bacterial community and microbially functional genes between crude oil and aqueous phases of oil production mixture was detected in Qinghai Oilfield. System research will carry out on the mechanism of microbial difference between crude oil and aqueous phases in distinct oil strata from Qinghai Oilfield by 454 sequencing, Real-time PCR, canonical correlation analysis, functional verification of aerobic and anaerobic hydrocarbon degradation simulated microcosms with differently microbial communities from crude oil and aqueous phase respectively, as well as the change of these microbial communities. In regard to the lack of isolates from crude oils, several novel isolation methods will be used to acquire diverse strains from crude oils. The performance of this study can provide new theoretical guidance and resource supply for efficient development of MEOR.

我国正面临能源短缺与大量石油资源未能有效开发的突出问题，因此利用油藏内丰富微生物资源开展的微生物提高石油采收率（MEOR）技术，以其可持续发展、经济有效的优点得到了广泛认可。但目前影响该技术发展的重要原因有二：忽视油藏微生物群落在原油与采出液水相中的差异，严重缺乏来源于原油的菌种资源。我们前期的初步研究发现，青海油田原油与采出液水相中细菌群落结构及功能基因存在差异，但整体微生物群落的差异机制还不得而知。因此，本项目拟以青海油田不同温度油藏原油、采出液水相及注入水为研究对象，运用高通量测序、实时荧光定量PCR、典型相关性分析及构建独立烃降解微环境等技术，定性、定量，并结合环境因子、功能作用分析两相中微生物群落的差异机制。同时，创新分离方法，突破原油来源菌种的高效分离培养，并筛选功能菌株。为加快油藏环境微生物资源在能源、微生物采油和生物修复等方面的高效应用奠定基础。

本项目围绕挖掘油藏环境丰富微生物资源，以期促进微生物提高石油采收率技术发展为目的，分析了青海油田及玉门油田、大庆油田不同温度油藏原油和采出液水相中细菌、古菌的群落结构及其差异，发现了假单胞菌及盐单胞菌分别在这些油藏原油及水相中的重要地位。并解析了青海油田油藏微生物群落与环境相关性，指出采出液矿化度、油藏温度及原油组分对微生物群落的显著影响。构建了代表油藏烃降解功能厌氧模拟体系，建立了适合原油来源微生物菌种的分离培养方法，获得了414株油藏环境好氧和厌氧细菌、古菌菌株，其中挖掘出潜在新物种26种，并用多种方法保藏代表菌株189株。目前已发表SCI学术论文2篇，并另有一篇SCI论文正在二次返修审稿中，后续还有正在撰写的论文会陆续投稿。本项目为目前限制微生物提高石油采收率技术发展的一大瓶颈问题给出了一定的解答，并提供了大量原油来源微生物资源，为后续功能菌株在生产中的高效应用打下了坚实的基础。