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一种新型嗜烟碱细菌节杆菌属aRF-1的分离、鉴定及其代谢途径

Isolation and characterization of a novel nicotinophilic bacterium, Arthrobacter sp. aRF-1 and its metabolic pathway.

作者信息

Ruan Aidong, Gao Yuan, Fang Chao, Xu Yaofei

机构信息

State Key Laboratory of Hydrology-Water Resources and Hydraulic Engineering, Hohai University, Nanjing, 210098, People's Republic of China.

出版信息

Biotechnol Appl Biochem. 2018 Nov;65(6):848-856. doi: 10.1002/bab.1682. Epub 2018 Aug 10.

Abstract

Nicotine is a potent parasympathomimetic stimulant and an important natural alkaloid mainly found in the Nicotiana genus of plants. It can directly threaten ecological security and human health in tobacco waste, wastewater, and other forms of tobacco production. Therefore, it is the basis of nicotine pollution prevention and of great application value to explore efficient and a wide range of nicotinophilic bacteria for tobacco industry and environmental protection. In this study, one nicotinophilic bacterium was isolated from the soil, which accumulated tobacco waste over 50 years at a Hefei cigarette factory. The strain was named aRF-1, which was identified as Arthrobacter sp. by analysis. The nicotine degradation tests showed that the optimum temperature for cell growth and metabolism of nicotine of Arthrobacter sp. aRF-1 was 30 °C, and the optimum initial pH value was about 7.0. Under the optimum experimented conditions, it can tolerance nicotine concentration as high as 8 g·L . The highest removal rate of nicotine was 93.8% in 72 H in nonsterilization contaminated soil by Arthrobacter sp. aRF-1. LC-MS/MS was used to analyze the nicotine metabolic intermediates of strain Arthrobacter sp. aRF-1. A total of nine major metabolites that were detected were able to metabolize nicotine along a variant pathway of pyridine and pyrrolidine, and there may be more than two nicotine metabolic pathways for Arthrobacter sp. aRF-1 through the analysis of the main intermediate products.

摘要

尼古丁是一种强效拟副交感神经兴奋剂,也是一种重要的天然生物碱,主要存在于烟草属植物中。它可以以烟草废料、废水及其他烟草生产形式直接威胁生态安全和人类健康。因此,探索高效且广泛的嗜烟碱细菌用于烟草工业和环境保护,是预防尼古丁污染的基础且具有重要应用价值。在本研究中,从合肥卷烟厂一处积累了50多年烟草废料的土壤中分离出一株嗜烟碱细菌。该菌株命名为aRF-1,经分析鉴定为节杆菌属。尼古丁降解试验表明,节杆菌属aRF-1细胞生长和尼古丁代谢的最适温度为30℃,最适初始pH值约为7.0。在最佳实验条件下,它能耐受高达8 g·L的尼古丁浓度。在非灭菌污染土壤中,节杆菌属aRF-1在72小时内对尼古丁的最高去除率为93.8%。采用液相色谱-串联质谱法(LC-MS/MS)分析节杆菌属aRF-1菌株的尼古丁代谢中间体。共检测到9种主要代谢产物,它们能够沿着吡啶和吡咯烷的不同途径代谢尼古丁,通过对主要中间产物的分析,节杆菌属aRF-1可能存在两条以上的尼古丁代谢途径。

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