质粒pT7降解三氯苯的功能研究

The function study on trichlorobenzene degradation by plasmid pT7

从以1,2,4-三氯苯为唯一碳源的降解菌--施氏假单胞菌(Pseudomonas Stutzeri)T7中提取到一个降解质粒,为研究该质粒的降解功能,将其转化到E.coli JM109中,转化子获得了降解1,2,4-三氯苯的能力.采用色谱/质谱联用(GC/MS)技术检测到转化子E.coli JM109(含质粒pT7)降解1,2,4-三氯苯过程中的部分中间产物分别为2,3,5-三氯-2,4-己二烯二酸、2,5-二氯-2,4-二烯-1,4-内酯-己酸、2,5-二氯-4-氧代-2-烯己二酸;在转化子中分别检测了三氯苯双加氧酶、脱氢酶、邻苯二酚1,2-双加氧酶的酶活性.

A degradation plasmid was drawn in Pseudomonas Stutzeri T7 using 1,2,4-trichlorobenzene （1,2,4-TCB） as the degradation bacteria of only carbon source. For studying the function of this plasmid, it was transformed in E. coli JM109, thus obtaing the ability to degrade 1,2,4-TCB. In the process of E. coli JM109 （containing plasmid pT7） degrading 1,2,4-TCB, the partial intermediate products were inspected and surveyed as 2,3,5-trichloromuconate, 2,5-dichloro-carboxymethylenebut-2-en-4-olide, and 2,5-dichloro-4-oxohex-2-enedioate respectively, adopting gas chromatography-mass spectrometry （GC-MS） technique. The enzyme activities of trichlorobenzene dioxygenase, dihydrodiol dehydrogenase, and catechol 1,2-dioxygenase, the enzymes were inspected and surveyed respectively.