Department of Environmental Systems Engineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan.
J Biosci Bioeng. 2012 Sep;114(3):306-11. doi: 10.1016/j.jbiosc.2012.04.010. Epub 2012 May 10.
Tris(2-chloroethyl) phosphate (TCEP), a flame retardant, is recently regarded as a potentially toxic and persistent environmental contaminant. We previously isolated TCEP-degrading bacterium, Sphingobium sp. strain TCM1, which, however, produced a toxic metabolite: 2-chloroethanol (2-CE). This study was undertaken to develop a detoxification technique of TCEP using strain TCM1 with a 2-CE-degrading bacterium: Xanthobacter autotrophicus strain GJ10. TCEP degradation by strain TCM1-resting cells was thermally stable for 30 min at 30 °C. It was optimal at 30 °C and at pH 8.5. In the optimum condition, TCM1 cells up to a final cell density of 0.8 at OD(660) in the reaction mixture were unable to hydrolyze the phosphotriester bonds of 10 μM TCEP completely. The addition of 50 μM Co(2+) to reaction mixture enhanced the hydrolysis and caused the complete hydrolysis at the cell density of 0.8. Strain GJ10 resting cells degraded 2-CE only slightly, which might be attributable to lack of coenzyme regeneration of enzymes involved in the degradation. In contrast, the growing cells degraded approximately 180 μM of 2-CE within 24 h. Based on these results, we designed a two-step TCEP detoxification reaction consisting of TCEP hydrolysis to 2-CE by strain TCM1-resting cells and the following degradation of the resulting 2-CE by strain GJ10-growing cells. The combined reaction completely detoxified 10 μM TCEP, and thus opens a way to microbial detoxification of the potential toxic, persistent organophosphorus compound.
三(2-氯乙基)磷酸酯(TCEP)是一种阻燃剂,最近被认为是一种具有潜在毒性和持久性的环境污染物。我们之前分离到了能够降解 TCEP 的细菌,即鞘氨醇单胞菌(Sphingobium sp.)TCM1 菌株,但该菌株会产生一种有毒的代谢物:2-氯乙醇(2-CE)。本研究旨在利用 TCEP 降解菌鞘氨醇单胞菌(Sphingobium sp.)TCM1 与 2-CE 降解菌黄单胞菌(Xanthobacter autotrophicus)GJ10 开发一种 TCEP 解毒技术。TCM1 菌株的休止细胞对 TCEP 的降解在 30°C 下 30 分钟内具有热稳定性。其最适温度为 30°C,最适 pH 值为 8.5。在最佳条件下,当反应混合物中 TCM1 细胞的最终 OD(660)达到 0.8 时,无法完全水解 10μM TCEP 的膦酸三酯键。向反应混合物中添加 50μM Co(2+)可增强水解作用,并使细胞密度为 0.8 时完全水解。GJ10 菌株的休止细胞仅略微降解 2-CE,这可能是由于参与降解的酶缺乏辅酶再生。相比之下,生长细胞在 24 小时内可降解约 180μM 的 2-CE。基于这些结果,我们设计了两步 TCEP 解毒反应,包括 TCM1 休止细胞将 TCEP 水解为 2-CE 和随后 GJ10 生长细胞降解生成的 2-CE。组合反应可完全解毒 10μM TCEP,从而为潜在有毒、持久性有机磷化合物的微生物解毒开辟了一条道路。