Mondragón-Parada M E, Ruiz-Ordaz N, Tafoya-Garnica A, Juárez-Ramírez C, Curiel-Quesada E, Galíndez-Mayer J
Laboratorio de Bioingeniería, Escuela Nacional de Ciencias Biológicas, IPN, Prol. Carpio y Plan de Ayala, Col. Santo Tomás, 11340 Mexico, DF, Mexico.
J Ind Microbiol Biotechnol. 2008 Jul;35(7):767-76. doi: 10.1007/s10295-008-0347-4. Epub 2008 Apr 8.
Using a successive transfer method on mineral salt medium containing simazine, a microbial community enriched with microorganisms able to grow on simazine was obtained. Afterwards, using a continuous enrichment culture procedure, a bacterial community able to degrade simazine from an herbicide formulation was isolated from a chemostat. The continuous selector, fed with a mineral salt medium containing simazine and adjuvants present in the commercial herbicide formulation, was maintained in operation for 42 days. Following the lapse of this time, the cell count increased from 5 x 10(5) to 3 x 10(8) CFU mL(-1), and the simazine removal efficiency reached 96%. The chemostat's bacterial diversity was periodically evaluated by extracting the culture's bacterial DNA, amplifying their 16S rDNA fragments and analyzing them by thermal gradient gel electrophoresis. Finally, a stable bacterial consortium able to degrade simazine was selected. By PCR amplification, sequencing of bacterial 16S rDNA amplicons, and comparison with known sequences of 16S rDNA from the NCBI GenBank, eight bacterial strains were identified. The genera, Ochrobactrum, Mycobacterium, Cellulomonas, Arthrobacter, Microbacterium, Rhizobium and Pseudomonas have been reported as common degraders of triazinic herbicides. On the contrary, we were unable to find reports about the ability of the genus Pseudonocardia to degrade triazinic compounds. The selected bacterial community was attached to a porous support in a concurrently aerated four-stage packed-bed reactor fed with the herbicide. Highest overall simazine removal efficiencies eta (SZ) were obtained at overall dilution rates D below 0.284 h(-1). However, the multistage packed bed reactor could be operated at dilution rates as high as D = 3.58 h(-1) with overall simazine removal volumetric rates R (v,SZ) = 19.6 mg L(-1) h(-1), and overall simazine removal specific rates R (X,SZ) = 13.48 mg (mg cell protein)(-1) h(-1). Finally, the consortium's ability to degrade 2-chloro-4,6-diamino-1,3,5-triazine (CAAT), cyanuric acid and the herbicide atrazine, pure or mixed with simazine, was evaluated in fed batch processes.
在含有西玛津的矿物盐培养基上采用连续转移法,获得了一个富含能在西玛津上生长的微生物的群落。之后,使用连续富集培养程序,从恒化器中分离出一个能够降解除草剂配方中西玛津的细菌群落。连续选择器以含有西玛津和商业除草剂配方中存在的助剂的矿物盐培养基为进料,持续运行42天。这段时间过后,细胞计数从5×10⁵增加到3×10⁸ CFU mL⁻¹,西玛津去除效率达到96%。通过提取培养物中的细菌DNA、扩增其16S rDNA片段并通过热梯度凝胶电泳进行分析,定期评估恒化器中的细菌多样性。最后,选择了一个能够降解西玛津的稳定细菌联合体。通过PCR扩增、细菌16S rDNA扩增子测序以及与NCBI GenBank中已知的16S rDNA序列进行比较,鉴定出了8种细菌菌株。据报道,慢生根瘤菌属、分枝杆菌属、纤维单胞菌属、节杆菌属、微杆菌属、根瘤菌属和假单胞菌属是三嗪类除草剂的常见降解菌。相反,我们未找到关于诺卡氏菌属降解三嗪类化合物能力的报道。所选细菌群落附着在一个多孔载体上,置于一个同时曝气的四级填充床反应器中,该反应器以除草剂为进料。在总稀释率D低于0.284 h⁻¹时,获得了最高的总体西玛津去除效率η(SZ)。然而,多级填充床反应器可以在高达D = 3.58 h⁻¹的稀释率下运行,总体西玛津去除体积速率R(v,SZ) = 19.6 mg L⁻¹ h⁻¹,总体西玛津去除比速率R(X,SZ) = 13.48 mg (mg细胞蛋白)⁻¹ h⁻¹。最后,在补料分批培养过程中评估了该联合体降解2-氯-4,6-二氨基-1,3,5-三嗪(CAAT)、氰尿酸以及纯的或与西玛津混合的除草剂阿特拉津的能力。