基于紫外-ARTP复合诱变选育高效MTBE降解突变菌株  

Breeding of Highly Efficient Methyl Tert-butyl Ether Biodegrading Mutant Strains Based on UV-ARTP Compound Mutagenesis

作　　者：赵盛 李玲玲 高翔 张昌运 王曰杰 ZHAO Sheng;LI Lingling;GAO Xiang;ZHANG Changyun;WANG Yuejie(State Key Laboratory of Chemical Safety,SINOPEC Research Institute of Safety Engineering Co.,Ltd.,Qingdao 266100,China;National Registration Center for Chemicals,Ministry of Emergency Management,Qingdao 266100,China;SINOPEC National Petrochemical Project Risk Assessment Technical Center Co.Ltd.,Qingdao 266100,China)

机构地区：[1]中石化安全工程研究院有限公司,化学品安全全国重点实验室,山东青岛266100 [2]应急管理部化学品登记中心,山东青岛266100 [3]中石化国家石化项目风险评估技术中心有限公司,山东青岛266100

出　　处：《环境科学与技术》2025年第2期88-95,共8页Environmental Science & Technology

基　　金：中国石化科技部项目(323092)。

摘　　要：传统人工驯化选育手段筛选效率低,为满足场地甲基叔丁基醚(MTBE)污染生物修复对高性能菌株的工业化需求,该文对出发菌株拉乌尔菌(Raoultella ornithinolytica QLM-1)进行紫外-常压室温等离子体(ARTP)复合诱变,为高效MTBE降解突变菌株的研发提供技术参考。结果表明,复合诱变最佳条件为:17 W紫外灯照射60 s,120 W ARTP辐照50 s,此条件下正突变率最高。通过高通量初筛和摇瓶培养复筛,得到遗传稳定的复合诱变优势菌株UAQ-39,相比出发菌株其降解率、比降解速率分别提高了52.65%、44.13%。菌株烷烃羟化酶、细胞色素P450酶活力增加,提示降解关键酶活性与MTBE生物降解进程强相关。该研究采用的紫外-ARTP复合诱变条件可显著提高菌株的MTBE降解性能,可为异生型污染物降解菌的开发提供技术应用参考。The screening efficiency of traditional artificial domestication breeding method is low.The original strain Raoultella ornithinolytica QLM-1 was subjected to UV-ARTP compound mutagenesis to obtain an efficient methyl tert-butyl ether(MTBE)degrading mutant strain,aiming at providing technical reference for the development of highly efficient MTBE degrading strains.Results showed that the optimal conditions of compound mutagenesis were 17 W ultraviolet lamp irradia-tion for 60 s,and 120 W ARTP irradiation for 50 s,and the positive mutation rate was the highest under this condition.Through high-throughput primary screening and shake flask re-screening,the genetically stable compound mutagenesis domi-nant strain UAQ-39 was obtained.Compared with the original strain,the degradation rate and specific degradation rate of UAQ-39 were increased by 52.65%and 44.13%,respectively.The activity of alkane hydroxylase and cytochrome P450 enzymes of UAQ-39 were significantly increased,indicating that the activities of key degradation enzymes were closely relat-ed to the biodegradation of MTBE.Research shows that the UV-ARTP compound mutagenesis conditions used in this study can significantly improve the MTBE degradation performance of the strain,and can provide technical reference for the devel-opment of heterogenic contaminant degrading bacteria.

关 键 词：甲基叔丁基醚 紫外诱变 常压室温等离子体诱变 复合诱变 生物修复 

分 类 号：X172[环境科学与工程—环境科学]