一株尼古丁降解菌株Arthrobacter sp.D4的分离鉴定及其降解代谢途径  

Isolation,identification,degradation pathway prediction of a nicotine-degrading strain Arthrobacter sp.D4

作　　者：邓帅军 张鸽 刘德水 貊志杰 史素娟 李志豪 马莉[4] 刘晓晖 张海波[3] 刘好宝[1] 王倩[1] DENG Shuaijun;ZHANG Ge;LIU Deshui;MO Zhijie;SHI Sujuan;LI Zhihao;MA Li;LIU Xiaohui;ZHANG Haibo;LIU Haobao;WANG Qian(Tobacco Research Institute of Chinese Academy of Agricultural Sciences,Qingdao 266101,Shandong,China;Graduate School of Chinese Academy of Agricultural Sciences,Beijing 100081,China;Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,Qingdao 266101,Shandong,China;Beijing Branch,Technology Center,Shanghai Tobacco Group Limited Company,Beijing 101121,China;Yichang Branch of Hubei Tobacco Company,Yichang 443000,Hubei,China;Shenzhen Tobacco Industry Limited Company,Shenzhen 518000,Guangdong,China)

机构地区：[1]中国农业科学院烟草研究所,山东青岛266101 [2]中国农业科学院研究生院,北京100081 [3]中国科学院青岛生物能源与过程研究所,山东青岛266101 [4]上海烟草集团有限责任公司技术中心北京工作站,北京101121 [5]湖北省烟草公司宜昌市公司,湖北宜昌443000 [6]深圳烟草工业有限责任公司,广东深圳518000

出　　处：《微生物学通报》2022年第9期3644-3656,共13页Microbiology China

基　　金：中国农业科学院烟草研究所横向项目(K2022-2-002P);国家自然科学基金(32170387)。

摘　　要：【背景】烟草在生产和加工中会产生高浓度的尼古丁废弃物,对环境造成较大的污染。【目的】筛选降解尼古丁的微生物菌种并解析其降解尼古丁的代谢途径,理解微生物如何降解尼古丁。【方法】用常规分离筛选方法、结合形态学观察和分子鉴定手段分离和鉴定菌株类别,进而利用单因素试验方法,通过设置不同的尼古丁浓度、温度和pH确定菌株降解尼古丁的最适发酵条件和降解率,利用气相色谱-质谱联用技术检测菌株在尼古丁降解过程中的主要代谢产物。【结果】获得一株以尼古丁为唯一碳源和氮源的节杆菌属(Arthrobacter)菌株,编号为D4;该菌株降解尼古丁的最适温度和pH分别为30.0°C和7.0;在1 g/L的尼古丁浓度下具备较快的尼古丁降解速率,培养18 h时尼古丁降解率可达到90%以上;尼古丁浓度≥4 g/L时菌株生长受到明显抑制;与目前报道的节杆菌属降解途径不同,该菌株降解尼古丁过程中产生了新的终产物N-甲基吡咯烷酮、可替宁及中间产物麦斯明。【结论】本研究分离鉴定到一株具有较快尼古丁降解速率的节杆菌,该菌株很可能存在新的尼古丁降解途径。[Background]High concentrations of nicotine waste are generated during the production and processing of tobacco,causing serious environmental pollution.[Objective]The purpose of this study is to screen out the microbial strain capable of degrading nicotine,and analyze the metabolic pathways of the strain in nicotine degradation.[Methods]We employed the conventional method for strain isolation and identified the target strain based on morphological characteristics and molecular evidence.The fermentation conditions including nicotine concentration,temperature,and pH were optimized by single factor tests with degradation rate as the indicator.The main metabolites of the strain in nicotine degradation were detected by gas chromatography-mass spectrometry.[Results]Arthrobacter sp.D4 was isolated,which could utilize nicotine as the sole carbon and nitrogen source.The optimal conditions of the strain for degrading nicotine were 30.0°C,pH 7.0,and nicotine concentration of 1 g/L,under which the degradation rate reached 90%within 18 h.The growth of the strain was significantly inhibited when the concentration of nicotine in the culture medium was greater than or equal to 4 g/L.Subsequent experiments revealed that the strain produced new end-products N-methylpyrrolidone and cotinine and the intermediate product myosmine,which were different from the reported degradation pathway of Arthrobacter sp.[Conclusion]In this study,an Arthrobacter sp.strain characterized by fast nicotine degradation was isolated and identified.Moreover,this strain may have a new nicotine degradation pathway.

关 键 词：尼古丁降解 节杆菌属 代谢产物 降解途径 

分 类 号：X172[环境科学与工程—环境科学] X795