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新分离的假单胞菌 CS3 及其代谢物对尼古丁的生物降解作用。

Biodegradation of nicotine by newly isolated Pseudomonas sp. CS3 and its metabolites.

机构信息

School of Life Sciences, Hunan University of Science and Technology, Xiangtan, China.

出版信息

J Appl Microbiol. 2012 Feb;112(2):258-68. doi: 10.1111/j.1365-2672.2011.05208.x. Epub 2012 Jan 3.

Abstract

AIMS

Isolation and characterization of nicotine-degrading bacteria with advantages suitable for the treatment of nicotine-contaminated water and soil and detection of their metabolites.

METHODS AND RESULTS

A novel nicotine-degrading bacterial strain was isolated from tobacco field soil. Based on morphological and physiochemical properties and sequence of 16S rDNA, the isolate was identified as Pseudomonas sp., designated as CS3. The optimal culture conditions of strain CS3 for nicotine degradation were 30°C and pH 7·0. However, the strain showed broad pH adaptability with high nicotine-degrading activity between pH 6·0 and 10·0. Strain CS3 could decompose nicotine nearly completely within 24 h in liquid culture (1000 mg L(-1) nicotine) or within 72 h in soil (1000-2500 mg kg(-1) nicotine) and could endure up to 4000 mg L(-1) nicotine in liquid media and 5000 mg kg(-1) nicotine in soil. Degradation tests in flask revealed that the strain had excellent stability and high degradation activity during the repetitive degradation processes. Additionally, three intermediates, 3-(3,4-dihydro-2H-pyrrol-5-yl) pyridine, 1-methyl-5-(3-pyridyl) pyrrolidine-2-ol and cotinine, were identified by GC/MS and NMR analyses.

CONCLUSIONS

The isolate CS3 showed outstanding nicotine-degrading characteristics such as high degradation efficiency, strong substrate endurance, broad pH adaptability, and stability and persistence in repetitive degradation processes and may serve as an excellent candidate for applications in the bioaugmentation process to treat nicotine-contaminated water and soil. Also, detection of nicotine metabolites suggests that strain CS3 might decompose nicotine via a unique nicotine-degradation pathway.

SIGNIFICANCE AND IMPACT OF THE STUDY

The advantage of applying the isolated strain lies in broad pH adaptability and stability and persistence in repetitive use, the properties previously less focused in other nicotine-degrading micro-organisms. The strain might decompose nicotine via a nicotine-degradation pathway different from those of other nicotine-utilizing Pseudomonas bacteria reported earlier, another highlight in this study.

摘要

目的

分离和鉴定具有适合处理尼古丁污染水和土壤的优点的尼古丁降解细菌,并检测其代谢物。

方法与结果

从烟草地土壤中分离到一株新型尼古丁降解细菌。根据形态学、生理生化特性和 16S rDNA 序列分析,该分离菌被鉴定为假单胞菌,命名为 CS3。菌株 CS3 降解尼古丁的最佳培养条件为 30℃和 pH7.0。然而,该菌株在 pH6.0-10.0 之间具有广泛的 pH 适应性和较高的尼古丁降解活性。在液体培养(1000mg/L 尼古丁)或土壤中(1000-2500mg/kg 尼古丁),菌株 CS3 可在 24h 内几乎完全降解尼古丁,在液体培养基中可耐受高达 4000mg/L 尼古丁,在土壤中可耐受高达 5000mg/kg 尼古丁。在摇瓶降解试验中发现,该菌株在重复降解过程中具有良好的稳定性和较高的降解活性。此外,通过 GC/MS 和 NMR 分析鉴定出 3 种中间产物,分别为 3-(3,4-二氢-2H-吡咯-5-基)吡啶、1-甲基-5-(3-吡啶基)吡咯烷-2-醇和可替宁。

结论

分离株 CS3 具有高效降解尼古丁、较强的底物耐受能力、广泛的 pH 适应性、在重复降解过程中的稳定性和持久性等突出的降解特性,可能是用于生物强化处理尼古丁污染水和土壤的优良候选菌株。同时,尼古丁代谢产物的检测表明,该菌株可能通过一种独特的尼古丁降解途径来降解尼古丁。

研究的意义和影响

该分离菌株的优势在于其在重复使用中的广泛 pH 适应性和稳定性,这是其他尼古丁降解微生物较少关注的特性。该菌株可能通过与之前报道的其他利用尼古丁的假单胞菌不同的尼古丁降解途径来降解尼古丁,这也是本研究的另一个亮点。

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