Department of Food Science and Microbiology, State University of Milan, Italy.
J Appl Microbiol. 2009 Apr;106(4):1111-21. doi: 10.1111/j.1365-2672.2008.04074.x. Epub 2009 Jan 15.
To isolate thiodiglycol (TDG)-degrading bacteria, the mustard gas hydrolysis product, and to characterize the metabolites formed and the enzymes involved in the degradation.
Two strains, identified as Achromobacter xylosoxydans G5 and Paracoccus denitrificans E4, isolated from a petroleum-contaminated soil, utilized TDG as sole carbon and sulfur source. During the degradation of TDG by strain E4 [(2-hydroxyethyl)thio] acetic acid (HETA), thiodiglycolic acid (TDGA) and bis-(2-hydroxyethyl)disulfide (BHEDS) were identified by gas chromatography-mass spectrometry analysis, while HETA and TDGA were identified for strain G5. Two-dimensional isoelectric focussing-gel electrophoresis (2-D IEF/SDS-PAGE) maps of protein extracts of P. denitrificans E4 grown on TDG showed a spot identified as a methanol dehydrogenase. Increased expression of a putative iscS gene, involved in sulfur assimilation, was observed in TDG-grown cells of A. xylosoxydans G5.
TDG degradation by P. denitrificans E4 occurred through two pathways: one involved cleavage of the C-S bond of HETA, yielding BHEDS and the other, oxidation of the alcoholic groups of TDG, yielding TDGA. The cleavage of the C-S bond of TDGA gave mercaptoacetic acid, further oxidized to acetate and sulfate.
Increased knowledge of TDG-degrading bacteria and the possibility of using them in a tailored-two-stage mustard gas destruction process.
分离硫代二甘醇(TDG)-降解菌,即芥子气水解产物,并对其代谢产物和参与降解的酶进行鉴定。
从石油污染土壤中分离到两株菌,鉴定为无色杆菌(Achromobacter xylosoxydans)G5 和脱氮副球菌(Paracoccus denitrificans)E4,它们可以将 TDG 作为唯一的碳源和硫源利用。通过气相色谱-质谱联用分析,发现 E4 菌株在降解 TDG 的过程中生成了[(2-羟乙基)硫]乙酸(HETA)、硫代二甘酸(TDGA)和双-(2-羟乙基)二硫(BHEDS);而 G5 菌株则生成了 HETA 和 TDGA。利用二维等电聚焦-凝胶电泳(2-D IEF/SDS-PAGE)分析 E4 菌株在 TDG 上生长的蛋白提取物图谱,发现其中一个斑点被鉴定为甲醇脱氢酶。在 A. xylosoxydans G5 的 TDG 生长细胞中,观察到参与硫同化的假定 iscS 基因的表达增加。
脱氮副球菌 E4 通过两种途径降解 TDG:一种途径涉及 HETA 的 C-S 键断裂,生成 BHEDS 和另一种途径,即 TDG 的醇基团氧化,生成 TDGA。TDGA 的 C-S 键断裂生成巯基乙酸,进一步氧化为乙酸和硫酸盐。
增加对 TDG 降解菌的了解,并有可能将其用于定制的两段芥子气销毁过程。