1,2-二氯乙烷降解菌群的富集及关键降解菌Ancylobacter sp. BL0的分离鉴定  

Enrichment of 1,2-dichloroethane Degrading Bacterial Consortium,and Isolation and Identification of Ancylobacter sp.BL0 of a Key Degrading Bacterial Strain

作　　者：张志梅 张彦猛 谢东明 杨秀云 王浪 左梓涵 吴志国[3] ZHANG Zhi-mei;ZHANG Yan-meng;XIE Dong-ming;YANG Xiu-yun;WANG Lang;ZUO Zi-han;WU Zhi-guo(Tianjin Lejin Bohai Chemical Co.,Ltd.,Tianjin 300452;Tianjin Dansheng Huan Safety Technology Co.,Ltd.,Tianjin 300457;Tianjin University of Science and Technology,Tianjin 300457)

机构地区：[1]天津乐金渤海化学有限公司,天津300452 [2]天津市丹晟环安科技有限公司,天津300457 [3]天津科技大学,天津300457

出　　处：《生物技术通报》2024年第8期288-298,共11页Biotechnology Bulletin

基　　金：校企合作技术开发项目(2023120021000787)。

摘　　要：【目的】1,2-二氯乙烷(1,2-DCA)的污染会危害到人体的生命健康和环境生态安全。为获得高效的1,2-DCA生物降解种质资源,研究菌群特征并探索从中分离筛选高效1,2-DCA降解菌株的规律和方法。【方法】从受1,2-DCA污染的土壤中富集培养能以1,2-DCA为唯一碳源和能源的降解菌群,通过气相色谱、紫外分光光度仪考察了不同批次菌群的生长和对1,2-DCA的降解情况,利用高通量测序分析不同批次富集液的物种多样性和相对丰度。通过16S rDNA基因序列分析鉴定菌株,利用气相色谱-质谱联用测定降解产物,分析菌株的降解途径。【结果】得到了降解菌群BG1,且实验结果显示第6-11批和15批富集菌群在24 h内对12.5 mg/L 1,2-DCA的降解率呈先增加后降低趋势,菌群生物量(OD_(600))可从0.03增长至0.095,但菌群中Ancylobacter sp.的相对丰度随着持续富集而降低。从第9批富集液中筛选出一株菌命名为BL0,鉴定为Ancylobacter sp.,能在6 h内降解120 mg/L 1,2-DCA,推测BL0降解1,2-DCA的途径为1,2-DCA水解为2-氯乙醇,2-氯乙醇再被氧化为氯乙酸,然后被彻底降解利用。【结论】本研究得到了高效降解1,2-DCA的菌群BG1和菌株Ancylobacter sp.BL0,同时发现1,2-DCA降解菌株的筛选分离需要的样品富集过程周期较长且微生物生长量较低,不适于过度富集。菌株Ancylobacter sp.BL0与富集菌群中的其他菌株之间以竞争关系为主。【Objective】The pollution of 1,2-dichloroethane(1,2-DCA)will seriously harm human health and environmental ecological safety.In order to obtain efficient 1,2-DCA biodegrading microorganism resources,the characteristics of bacterial flora were studied,and then the rules and methods for separating and screening efficient 1,2-DCA degrading strains were explored.【Method】The bacteria utilizing 1,2-DCA as the sole carbon source and energy were enriched and cultured from soil contaminated by 1,2-DCA.The growth of bacteria and degradation of 1,2-DCA in different batches were investigated by ultraviolet spectrophotometry and gas chromatography respectively.The species diversity and relative abundance of different enriched solution were analyzed by high-throughput sequencing.The strain was identified by 16S rDNA gene sequence analysis.The degraded products were determined by gas chromatography-mass spectrometry,and the degradation pathway was analyzed.【Result】Bacterial consortium BG1 for degradation was gained,and the experimental results showed that the degrading rate of 12.5 mg/L 1,2-DCA by enriched bacterial consortium in batch 6-11 and 15 increased at the beginning and then decreased within 24 h,and the microbial biomass(OD_(600))increased from 0.03 to 0.095,but the relative abundance of Ancylobacter sp.in the flora decreased with continuous enrichment process.A strain named BL0 was screend from the batch 9 of enrichment solution and identified as Ancylobacter sp.,which degraded 120 mg/L 1,2-DCA within 6 h.It was inferred that the metabolic pathway of degradation of 1,2-DCA by BL0 was hydrolysis of 1,2-DCA to 2-chloroethanol at first,and 2-chloroethanol was oxidized to chloroacetic acid,which was then completely degraded and utilized.【Conclusion】Highly efficient 1,2-DCA degrading bacterial consortium BG1 and strain Ancylobacter sp.BL0 are obtained,and it is found that a long period of sample enrichment is required for screening and separation of 1,2-DCA degrading strains during the process of sample enri

关 键 词：1 2-二氯乙烷 生物降解 菌群富集 降解菌群 Ancylobacter 群落互作 代谢途径 

分 类 号：X172[环境科学与工程—环境科学]