BACKGROUND: Surfactant-enhanced remediation (SER) has been widely applied to remove petroleum hydrocarbon (PH) pollutants, but there are few studies on the mechanism through which surfactants affect the hydrocarbon metabolite ability of microorganism. RESULTS: In this work, combination bacteria and surfactant were selected to remove PH, and the degradation bottleneck concentration of single-bacteria was 10 g L -1 according to the PH degradation rate. However, the degradation efficient of mixed-bacteria was further increased after adding surfactants. Among them, the PH degradation rate of Kocuria rosea + Bacillus odyssey and Microbacterium. sp + B. odyssey at the bottleneck concentration had been respectively reached to 81.58 ± 6.29% with tween-80 and 88.35 ± 7.58% with rhamnolipid. Compared with no surfactant, the relative abundance of global overmaps, amino acid metabolism, and carbohydrate metabolism was increased in adding tween-80 condition. However, when rhamnolipid was added, the relative abundance of ABC transporters, two-component system, and bacteria chemotaxis was clearly increased, indicating an improvement of transportation, absorption, and degradation of PH. Additionally, the gene abundance of alkB and nah was also significantly increased with above condition. Meanwhile, the gene abundance of alkB and nah was also significantly enhanced by above condition.CONCLUSION: This study offers new insights into the functional changes of mixed-bacteria during PH degradation in different surfactant-enhanced systems.