Functional Study on the Degradation of Phenanthrene by Recombinant pET-28a-C12O/E.coli
Polycyclic aromatic hydrocarbon pollution is inevitable in the process of petroleum and shale oil extrac-tion.Taking the polycyclic aromatic hydrocarbon phenanthrene as the standard degradation substrate,the oil de-grading bacteria HL-1(Acinetobacter calcoaceticus)isolated from Daqing Petroleum No.3 Oil Production Plant was used as the standard degradation substrate.Using the DNA of strain HL-1 as a template,catechol 1,2 dioxyge-nase(C12O)was obtained through PCR amplification.C12O was connected to pET-28a through gene cloning and successfully expressed in E.coli BL21.At a temperature of 30℃,a constant shaker speed of 150r/min,an initial culture pH of 7.0,and a high concentration of 2000mg/L phenanthrene,after 7 days of degradation,the average degradation rate was 15.9％.Therefore,genetic engineering technology has good application prospects in dealing with the pollution of polycyclic aromatic hydrocarbons.
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