利用油平板筛选法和排油圈法从辽河某油田的土壤样品中分离筛选高效的生物表面活性剂产生菌,通过16S rRNA基因序列分析对所筛选菌株进行鉴定。采用L16(45)正交实验对所筛选菌株合成生物表面活性剂的条件进行优化。比较5种提取方法对发酵液中生物表面活性剂的提取效果。共分离得到17株菌,其中菌株A3、As和Y产表面活性剂的能力最强。16S rRNA基因序列分析表明,这3株菌均为假单胞菌属(Pseudomonas)。菌株A3、As、Y和A3-As混合菌的生物表面活性剂提取量较优化前有明显的提高,其产量分别是优化前的8.4倍、6.4倍、5.6倍和5.7倍,其中,菌株As的生物表面活性剂产量最高,可达20.55 g/L。对于3种纯菌和A3-As混合菌而言,5种提取方法中均以CHCl_3萃取的量最高;而对其他混合菌,5种提取方法的差别并不显著。菌株A3、As、Y均属于高效的生物表面活性剂产生菌,具有很好的应用前景。 The oil-plate screening method and oil drain circle method were used to isolate and screen the high-efficient biosurfactant-producing bacteria from the soil samples of an oilfield in Liaohe. The selected strains were identified by 16S rRNA gene sequence analysis. L16 (45) orthogonal experiment was used to optimize the biosurfactant synthesis conditions of the selected strains. The effects of five extraction methods on the extraction of biosurfactants from the fermentation broth were compared. Seventeen strains were isolated, of which strain A3, As and Y had the strongest ability to produce surfactants. 16S rRNA gene sequence analysis showed that these three strains are Pseudomonas. The biosurfactant extracts of strain A3, As, Y and A3-As were significantly higher than those before optimization, and their yields were 8.4, 6.4, 5.6 and 5.7 times before optimization, respectively. Among them, strain As The highest biosurfactant production, up to 20.55 g / L. For the three kinds of bacteria and A3-As mixed bacteria, the five extraction methods were the highest amount of CHCl_3 extraction; and other mixed bacteria, five kinds of extraction methods, the difference is not significant. Strain A3, As, Y are efficient biosurfactant-producing bacteria, has good application prospects.