Satapute Praveen, Kaliwal Basappa
Department of Microbiology and Biotechnology, Karnatak University, Dharwad, Karnataka, India.
Department of Microbiology and Biotechnology, Davangere University, Davangere, Karnataka, India.
3 Biotech. 2016 Jun;6(1):110. doi: 10.1007/s13205-016-0429-3. Epub 2016 May 11.
The isolation of propiconazole (PCZ) degrading bacterium BBK_9 strain was done from paddy soil, and it was identified as Burkholderia sp. based on the morphological characteristics and biochemical properties combined with 16S rRNA gene sequencing analysis. It has been seen that the factors such as temperature and pH influence the biodegradation process. The role of plasmid was studied in the degradation process by plasmid curing method. The PCZ acts as the sole carbon source and as energy substrate which can be utilized by the strain for its growth in Mineral salt medium and degraded 8.89 µg ml of PCZ at 30 °C and pH 7 within 4 days. During the bioconversion process of PCZ, three metabolite were formed such as 1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl) ethanone, 1-[2-(4-chlorophenyl) ethyl]-1H-1,2,4-triazole and 1-ethyl-1H-1,2,4-triazole. The LD50 value of BBK_9 strain was determined with acridine orange which resulted in 40 µg ml at cell density of 0.243 at 660 nm. Furthermore, plasmid curing was done using LD50 concentration and from that three plasmids got cured in the sixth generation. It was found that, cured strain was able to degrade 7.37 µg ml of PCZ, indicating the plasmid encoded gene were not responsible for the PCZ degradation. On the source of these outcomes, strain BBK_9 can be used as potential strain for bioremediation of contaminated sites.
从稻田土壤中分离出了降解丙环唑(PCZ)的BBK_9菌株,并根据形态特征、生化特性以及16S rRNA基因测序分析将其鉴定为伯克霍尔德菌属。研究发现温度和pH等因素会影响生物降解过程。通过质粒消除法研究了质粒在降解过程中的作用。丙环唑作为唯一碳源和能量底物,该菌株可在无机盐培养基中利用其生长,并在30℃和pH 7的条件下于4天内降解8.89μg/ml的丙环唑。在丙环唑的生物转化过程中,形成了三种代谢产物,即1-(2,4-二氯苯基)-2-(1H-1,2,4-三唑-1-基)乙酮、1-[2-(4-氯苯基)乙基]-1H-1,2,4-三唑和1-乙基-1H-1,2,4-三唑。用吖啶橙测定BBK_9菌株的LD50值,在660nm处细胞密度为0.243时,结果为40μg/ml。此外,使用LD50浓度进行质粒消除,第六代中有三个质粒被消除。结果发现,消除质粒的菌株能够降解7.37μg/ml的丙环唑,这表明质粒编码的基因与丙环唑降解无关。基于这些结果,BBK_9菌株可作为污染场地生物修复的潜在菌株。