尼古丁降解菌的筛选鉴定及降解特性的研究
摘要
本文利用富集筛选的方法从杭州农药厂的活性污泥中分离得到两株尼古丁高效降解菌,分别命名为HZN1和TW5,经过对这两株菌的生理生化特性、形态特征和16S rDNA序列同源性分析,将其分别鉴定为申氏杆菌属(Shinella sp.)和假单胞属(Pseudomonas sp.)。
研究表明,菌株HZN1和TW5都能以尼古丁为唯一碳源、氮源和能源生长,能在质量浓度为4.0 g/L的尼古丁基础盐培养基中生长,菌株生长和降解尼古丁的最适温度均为30℃,最适pH值为6.5~7.5。菌株HZN1分别能在4 h、9 h、12 h内将初始浓度分别为0.2、0.5、1.0 g/L的尼古丁降解接近完全,而外加碳源葡萄糖会减缓HZN1降解尼古丁的速率;温度和pH值对HZN1的降解效率影响不大;菌株TW5分别能在6 h和9 h内将初始浓度分别为0.2和0.5 g/L的尼古丁降解接近完全,当尼古丁质量浓度上升为1.5 g/L时,降解效率受到明显影响,12 h内只能降解70%的尼古丁,温度和pH值对TW5的降解效率影响较大。
进一步利用气质联用仪(GC-MS)研究了菌株HZN1降解尼古丁的代谢产物,根据检测到的三个产物:麦思明、可铁林和尼古提林,推测了菌株HZN1在降解尼古丁过程中可能存在的降解途径。
菌株HZN1和TW5具有在恶劣环境中处理高浓度尼古丁废水的潜力,其中HZN1所属的申氏杆菌属(Shinella sp.)还未见报道有能够降解尼古丁的菌种,我们的研究为生物降解尼古丁提供了新的依据。
The nicotine-degrading bacterium HZN1 and TW5 were isolated from activated sludge of Hangzhou pesticide factory and identified as Shinella sp. and Pseudomonas sp. based on their physiological characteristics and analysis of 16S rDNA gene, respectively.
Strain HZN1 and TW5 are capable of using nicotine as the sole source of carbon, nitrogen and energy, and can growth on the mineral salts medium with nicotine concentration 4.0 g/L. The optimum temperature and pH for strain HZN1 and TW5 growth and nicotine degradation were 30℃and 7.0, respectively. Strain HZN1 could degrade approximately 100% of 0.2, 0.5 1.0 g/L of nicotine within 4, 9, 12 h, respectively; the degradation of nicotine was slightly slower with the presence of glucose; strain HZN1 could degrade nicotine efficiently in a relatively broad range of temperature (23, 30, 37℃) and pH values (5.5 ~ 9.0). Strain TW5 could degrade approximately 100% of 0.2, 0.5 g/L of nicotine within 6, 9 h, respectively; it could only degrade 70% of 1.5 g/L of nicotine at 12 h; temperature and pH have high effects on nicotine degradation by strain TW5.
Three intermediate metabolites were produced by the stain HZN1 and identified as cotinine, myosmine and nicotyrine using gas chromatography-mass spectrometry (GC-MS). And possible nicotine-degrading pathway of strain HZN1 was presumed.
These results indicate that the strain HZN1 and TW5 could potentially be useful for the remediation of nicotine pollution in mal-condition. This is the first report of nicotine-degrading strain from the genus of Shinella. Our findings would provide a new insight into the biodegradation of nicotine.
研究表明,菌株HZN1和TW5都能以尼古丁为唯一碳源、氮源和能源生长,能在质量浓度为4.0 g/L的尼古丁基础盐培养基中生长,菌株生长和降解尼古丁的最适温度均为30℃,最适pH值为6.5~7.5。菌株HZN1分别能在4 h、9 h、12 h内将初始浓度分别为0.2、0.5、1.0 g/L的尼古丁降解接近完全,而外加碳源葡萄糖会减缓HZN1降解尼古丁的速率;温度和pH值对HZN1的降解效率影响不大;菌株TW5分别能在6 h和9 h内将初始浓度分别为0.2和0.5 g/L的尼古丁降解接近完全,当尼古丁质量浓度上升为1.5 g/L时,降解效率受到明显影响,12 h内只能降解70%的尼古丁,温度和pH值对TW5的降解效率影响较大。
进一步利用气质联用仪(GC-MS)研究了菌株HZN1降解尼古丁的代谢产物,根据检测到的三个产物:麦思明、可铁林和尼古提林,推测了菌株HZN1在降解尼古丁过程中可能存在的降解途径。
菌株HZN1和TW5具有在恶劣环境中处理高浓度尼古丁废水的潜力,其中HZN1所属的申氏杆菌属(Shinella sp.)还未见报道有能够降解尼古丁的菌种,我们的研究为生物降解尼古丁提供了新的依据。
The nicotine-degrading bacterium HZN1 and TW5 were isolated from activated sludge of Hangzhou pesticide factory and identified as Shinella sp. and Pseudomonas sp. based on their physiological characteristics and analysis of 16S rDNA gene, respectively.
Strain HZN1 and TW5 are capable of using nicotine as the sole source of carbon, nitrogen and energy, and can growth on the mineral salts medium with nicotine concentration 4.0 g/L. The optimum temperature and pH for strain HZN1 and TW5 growth and nicotine degradation were 30℃and 7.0, respectively. Strain HZN1 could degrade approximately 100% of 0.2, 0.5 1.0 g/L of nicotine within 4, 9, 12 h, respectively; the degradation of nicotine was slightly slower with the presence of glucose; strain HZN1 could degrade nicotine efficiently in a relatively broad range of temperature (23, 30, 37℃) and pH values (5.5 ~ 9.0). Strain TW5 could degrade approximately 100% of 0.2, 0.5 g/L of nicotine within 6, 9 h, respectively; it could only degrade 70% of 1.5 g/L of nicotine at 12 h; temperature and pH have high effects on nicotine degradation by strain TW5.
Three intermediate metabolites were produced by the stain HZN1 and identified as cotinine, myosmine and nicotyrine using gas chromatography-mass spectrometry (GC-MS). And possible nicotine-degrading pathway of strain HZN1 was presumed.
These results indicate that the strain HZN1 and TW5 could potentially be useful for the remediation of nicotine pollution in mal-condition. This is the first report of nicotine-degrading strain from the genus of Shinella. Our findings would provide a new insight into the biodegradation of nicotine.
引文
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