Study On Isolation And Characterization Of Nicotine Degrading Bacterium Strain Pseudomonas Putida ZB-16A

Nicotine is the principal alkaloid in most of Nicotinana species. It is regarded as one of malignant environmental pollutants because of harmful effects on the healthy of human beings. China is the biggest tobacco production and consumption country. The nicotine-degradation bacteria isolation and nicotine degradation study provide theoretical basis for the application of bacteria on treatment of nicotine pollution.1 .Isolation and screening of nicotine-degradating bacteria58 samples were collected from soils of tobacco planting in areas Zi Bo city (Zi Bo area and Yi Yuan village, Zi Bo city), Shan Dong province and Waste in Wuhan Tobacco Company. A total of 152 pure isolates were obtained from collecting samples. These isolates were all analyzed for their nicotine-degrading activities using a medium containing 0.15% nicotine as the sole carbon and nitrogen source. The strains have different capacity for nicotine degradation. 22 nicotine-degradation strains were obtained from 27 samples collected in Wuhan Tobacco Company. 76 nicotine-degradation strains were gotten from 19 samples that came from ZiBo aera in ZiBo city, ShanDong province. 12 samples collected from ZiBo city YiYuan village ShanDong province contained 54 nicotine-degradation strains. The tobacco planting soil (Zi Bo area and Zi Bo city Yi Yuan village) contained more nicotine-degradation strains than the waste in Wuhan Tobacco Company. Among the isolates, One isolate ZB-16A screened from tobacco planting soils (ZiBo city, Zibo area, ShanDong province) showed good growth state when the nicotine concentration was 4000mg/L.2 Identification of nicotine-degradation isolate ZB-16ABy the morphological observation, physiological and structural characteristics, comparison of its 16S rDNA and phylogenesis tree analysis, the strain ZB-16A was identified as Pseudomonas putida ZB-16A.3 Study on degradation characteristics of P. putida ZB-16AGrowth state and characteristics were detected in this research. P. putida ZB-16A showed very high capacity for nicotine degradation. The optimal temperature for nicotine metabolism was 25℃, although this strain could exhaust 3000mg/L nicotine when temperature ranged from 10℃to 40℃. This means this strain possessed good temperature adaptability. ZB-16A appeared relative fast cell growth and nicotine degradation with temperature of 40℃and pH 6.5-7.5. ZB-16A could tolerate 6000 mg/L nicotine and all the nicotine was consumed after 48h while the temperature and initial pH was 30℃and pH 7.0, respectively. ZB-16A growth and nicotine metabolism was inhibited in nicotine concentration 7000mg/L.