Microbial enhanced oil recovery (MEOR) is a promising technology for oil field extraction. This study investigated a co-culture system of Pseudomonas aeruginosa and Bacillus subtilis to increase MEOR efficacy. We analyzed bacterial growth, biosurfactant production, and crude oil emulsified performance under different inoculation ratios. Compared to single cultures, the co-culture system showed superior growth and functional expression, with an optimal inoculation ratio of 1:1. Quantitative assessments of the cell numbers and biosurfactant production during the co-culture revealed that rapid B. subtilis proliferation in early stages significantly stimulated P. aeruginosa growth. This interaction increased cell density and rhamnolipid production by 208.05% and 216.25%, respectively. The microscopic etching model displacement results demonstrated enhanced emulsification and mobilization of crude oil by the co-culture system, resulting in 94.48% recovery. A successful field application in a block-scale reservoir increased cumulative oil production by 3.25 × 103 t. An analysis of microbial community structure and function in different phases revealed that after co-culture system injection, Pseudomonas became the dominant genus in the reservoir community, with an average abundance of 24.80%. Additionally, the abundance of biosurfactant-producing and hydrocarbon-degrading bacteria increased significantly. This research and the application of the P. aeruginosa and B. subtilis co-culture system provide novel insights and strategies for MEOR.
Keywords: biosurfactant production; co-culture; cooperation mechanism; field trial; microbial community; microbial enhanced oil recovery; residual oil mobilization.