Microbial strains capable of degrading petroleum hydrocarbons were isolated from the Yellow River Delta and screened for bio-surfactant production. The bio-surfactant-producing characteristics of the isolates were evaluated, and all the isolates which could produce bio-surfactant were identified by 16S rRNA gene sequencing. The results showed that the isolates belong to Bacillussp. (72%), Ochrobactrumsp. (0.16%), Brevundimonassp. (0.06%) and Brevibacteriumsp. (0.06%). The biodegradability of crude oil, gasoline, diesel oil and other hydrocarbons by microbial strains were studied, among which the biodegrading ability of strain P1 and strain P19 is higher than other strains. Both strains P1 and P19 can degrade n-hexane and n-hexadecane effectively and have wide substrate extensiveness. In addition, Ni promoted the biodegradability of toluene by both strain P1 and strain P19, while Pb inhibited the growth of strain P19 and decreased its ability to biodegrade toluene. The studies revealed that microbes including strain P1 and strain P19 can be utilized in bioremediation of co-contaminated water with petroleum and heavy metals including Ni and Pb.