重组体pET-28a-alkB/E.coli降解页岩油泥石油烃的功能研究  

Biodegradation Function of the Petroleum Hydrocarbon in Shale Oily Sludge by the Recombinant pET-28a-alkB/E.coli

作　　者：李雪菲 句泽林 齐婷婷 郑乾璐 李喜梅 余丽芸[1] LI Xuefei;JU Zelin;QI Tingting;ZHENG Qianlu;LI Ximei;YU Liyun(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,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

出　　处：《山东化工》2024年第19期239-243,共5页Shandong Chemical Industry

基　　金：黑龙江省创新创业训练项目(编号S202310223073):烷烃单加氧酶alkB基因的大肠杆菌构建及应用;省科技厅重点研发计划项目(编号GA21C028):页岩油开采产生污油泥的降解菌株体系构建及其应用技术。

摘　　要：为了降解页岩油泥中的石油烃,构建基因工程菌,表达石油烃降解关键酶以提高油泥的降解效果。以Pseudomonas qingdaonensis基因组为模板扩增alkB基因,连接至载体pET-28a,转化至E.coli BL21(DE3)中,SDS-PAGE和Western-blot鉴定表达的外源蛋白。pET-28a-alkB/E.coli处理原油和页岩油泥,采用气相色谱法评估降解效果。发现alkB基因在大肠杆菌中能表达外源蛋白,且14 d原油及页岩油泥的降解率分别为47.5%和47.1%,表明重组体pET-28a-alkB/E.coli具备降解页岩油泥石油烃的功能。In order to degrade petroleum hydrocarbons in shale oily sludge,genetically engineered bacteria were constructed to express key enzymes for petroleum hydrocarbon degradation in order to improve the degradation of oil sludge.The alkB gene was amplified using the Pseudomonas qingdaonensis genome as a template,ligated into the vector pET-28a,transformed into E.coli BL21(DE3),and the expressed exogenous proteins were identified by SDS-PAGE and Western-blot.Treat crude oil and shale sludge by pET-28a-alkB/E.coli and evaluate degradation by gas chromatography.The alkB gene was found to express exogenous proteins in Escherichia coli and the degradation rates of 14 d crude oil and shale sludge were 47.5%and 47.1%,respectively,suggesting that the recombinant pET-28a-alkB/E.coli possesses the function of degrading petroleum hydrocarbons from shale sludge.

关 键 词：烷烃单加氧酶alkB 生物降解 E.coli BL21(DE3) 页岩油污泥 气相色谱法 

分 类 号：X742[环境科学与工程—环境工程]