Sharp Jonathan O, Wood Thomas K, Alvarez-Cohen Lisa
Department of Civil and Environmental Engineering, University of California at Berkeley, Berkeley, California 94720-1710, USA.
Biotechnol Bioeng. 2005 Mar 5;89(5):608-18. doi: 10.1002/bit.20405.
The water contaminant N-nitrosodimethylamine (NDMA) is a probable human carcinogen whose appearance in the environment is related to the release of rocket fuel and to chlorine-based disinfection of water and wastewater. Although this compound has been shown to be biodegradable, there is minimal information about the organisms capable of this degradation, and little is understood of the mechanisms or biochemistry involved. This study shows that bacteria expressing monooxygenase enzymes functionally similar to those demonstrated to degrade NDMA in eukaryotes have the capability to degrade NDMA. Specifically, induction of the soluble methane monooxygenase (sMMO) expressed by Methylosinus trichosporium OB3b, the propane monooxygenase (PMO) enzyme of Mycobacterium vaccae JOB-5, and the toluene 4-monooxygenases found in Ralstonia pickettii PKO1 and Pseudomonas mendocina KR1 resulted in NDMA degradation by these strains. In each of these cases, brief exposure to acetylene gas, a suicide substrate for certain monooxygenases, inhibited the degradation of NDMA. Further, Escherichia coli TG1/pBS(Kan) containing recombinant plasmids derived from the toluene monooxygenases found in strains PKO1 and KR1 mimicked the behavior of the parent strains. In contrast, M. trichosporium OB3b expressing the particulate form of MMO, Burkholderia cepacia G4 expressing the toluene 2-monooxygenase, and Pseudomonas putida mt-2 expressing the toluene sidechain monooxygenase were not capable of NDMA degradation. In addition, bacteria expressing aromatic dioxygenases were not capable of NDMA degradation. Finally, Rhodococcus sp. RR1 exhibited the ability to degrade NDMA by an unidentified, constitutively expressed enzyme that, unlike the confirmed monooxygenases, was not inhibited by acetylene exposure.
水中污染物N-亚硝基二甲胺(NDMA)是一种可能的人类致癌物,其在环境中的出现与火箭燃料的释放以及水和废水的氯基消毒有关。尽管已证明这种化合物可生物降解,但关于能够进行这种降解的生物体的信息极少,对其涉及的机制或生物化学也知之甚少。本研究表明,表达与真核生物中已证明可降解NDMA的单加氧酶功能相似的单加氧酶的细菌具有降解NDMA的能力。具体而言,诱导甲基弯曲菌OB3b表达的可溶性甲烷单加氧酶(sMMO)、母牛分枝杆菌JOB-5的丙烷单加氧酶(PMO)以及皮氏罗尔斯通氏菌PKO1和门多萨假单胞菌KR1中发现的甲苯4-单加氧酶,会导致这些菌株降解NDMA。在每种情况下,短暂暴露于乙炔气体(某些单加氧酶的自杀底物)会抑制NDMA的降解。此外,含有源自PKO1和KR1菌株中发现的甲苯单加氧酶的重组质粒的大肠杆菌TG1/pBS(Kan)模仿了亲本菌株的行为。相比之下,表达MMO颗粒形式的甲基弯曲菌OB3b、表达甲苯2-单加氧酶的洋葱伯克霍尔德菌G4以及表达甲苯侧链单加氧酶的恶臭假单胞菌mt-2均不能降解NDMA。此外,表达芳香族双加氧酶的细菌也不能降解NDMA。最后,红球菌RR1表现出通过一种未鉴定的组成型表达酶降解NDMA的能力,与已确认的单加氧酶不同,该酶不受乙炔暴露的抑制。