利用正交法分离鉴定苯胺降解菌及对其降解特性的研究

摘要

[ Objective ] The aim was to isolate and identify aniline degrading bacterium with orthogonal test, and study its degradation characteristics. [ Method] Three aniline degrading bacterium were obtained from the soil sample which was collected from the circumambience of drainage ditch in a printing plant by isolation and screening, and the physical and chemical properties were determined, 16s rDNA sequence was analyzed. The effects of 5 factors included temperature, pH value, the anilines concentration, incubation time (hours) and the rotate speed on degradation rate of aniline degrading bacterium were examined. Regarding the degradation rate of aniline as the index, choosing temperature, pH value, the anilines concentration, incubation time (hours) and the rotate speed as factors, the degradating condition were optimized by orthogonal test. [ Result ] Three degrading bacterium were identified as Pseudomonas sp. , Acinetobacter gerneri and Delftia sp. ,named as AN-Y, AN-B and AN-H. The optimum temperature and pH value for growth and aniline degradation of three aniline degrading bacterium were 30 ℃ and 7.0, while they were capable of growing on the aniline substrate with the most consistency of aniline reached to 3 300,2 000 and 3 000 mg/L. The most degrading conditions of temperature was 30 ℃ , pH value was 8.0, the anilines concentration was 2 000 mg/L, rotate speed was 160 r/min and incubation time (hours) was 72 h. The variance analysis showed that the amount of aniline had significant effect on the degradation efficiency of degrading bacterium. [ Conclusion ] The study provides theoretical basis for the bacilliculture of aniline degrading bacterium.%[目的]利用正交法分离鉴定苯胺降解菌,并对其降解特性进行研究.[方法]在某印刷厂排污口周围土壤样品中分离筛选出3株苯胺降解菌,测定其生理生化特性,并对其16s rDNA序列进行分析,考察不同温度、pH、摇床转速、培养时间及苯胺浓度对一株高效苯胺降解菌降解率的影响,并以苯胺降解率为指标,选择温度、pH、摇床转速、培养时间及苯胺浓度等因素进行正交试验设计,优化该菌株对苯胺的降解条件.[结果]3株苯胺降解菌AN-Y、AN-B和AN-H分别鉴定为Pseudomonas sp.、Acinetobacter gerneri和Delftia sp.;生长和降解效果的最适温度为30℃,最适pH为7.0,并可分别在苯胺浓度为3 300、2 000和3 000 mg/L基础培养基上生长;耐受能力最强的降解菌AN-Y其最佳降解条件为温度35 ℃、pH 8.0、苯胺浓度2 000 mg/L、摇床转速160 r/min、培齐时间72 h;方差分析表明,苯胺浓度对降解菌降解苯胺的效果影响最为显著.[结论]研究结果为苯胺降解菌的培养研究提供了参考.