重组体pET-28a-C12O/E.coli降解菲的功能研究  

Functional Study on the Degradation of Phenanthrene by Recombinant pET-28a-C12O/E.coli

作　　者：王超 邓爽 李丽阳[1] 曹迪[1] 侯喜林[1] 余丽芸[1] 毛立丰 沈江川 Wang Chao;Deng Shuang;Li Liyang;Cao Di;Hou Xilin;Yu Liyun;Mao Lifeng;Shen Jiangchuan(Key Laboratory of Environmental Microbiology and Recycling in Cold Regions of Heilongjiang Province,College of Life Science and Technology,Heilongjiang Bayi Agricultural Reclamation University,Daqing 163000,Heilongjiang,China;Daqing Yilai Inspection and Testing Technology Service Co.,Ltd.,Daqing 163000,Heilongjiang,China)

机构地区：[1]黑龙江八一农垦大学,生命科学技术学院,黑龙江省寒区环境微生物及循环利用重点实验室,黑龙江大庆163000 [2]大庆亿莱检验检测技术服务有限公司,黑龙江大庆163000

出　　处：《绿色科技》2024年第16期266-272,280,共8页Journal of Green Science and Technology

基　　金：黑龙江省科技厅重点研发计划项目(编号:GA21C028)。

摘　　要：多环芳烃污染在石油及页岩油开采过程中不可避免,以多环芳烃菲作为标准降解底物,利用实验室初期从大庆石油采油三厂中分离得到的石油降解菌HL-1(Acinetobacter calcoaceticus)。以菌株HL-1为模板,通过PCR扩增的方法得到邻苯二酚1,2双加氧酶(C12O)。将C12O通过连接到表达载体pET-28a,在E.coli BL21中得到成功表达。在温度为30℃、摇床转速恒定在150 r/min,培养的初始pH值为7.0,菲浓度为2000 mg/L的高浓度下,经过7 d的降解,其降解率为15.9%。因此,通过基因工程技术在应对多环芳烃的污染方面有着较好的应用前景。Polycyclic aromatic hydrocarbon pollution is inevitable in the process of petroleum and shale oil extraction.Taking the polycyclic aromatic hydrocarbon phenanthrene as the standard degradation substrate,the oil degrading 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 dioxygenase(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°C,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.

关 键 词：多环芳烃 菲 BL21 邻苯二酚1 2双加氧酶 

分 类 号：X172[环境科学与工程—环境科学]