Wackett L P, Sadowsky M J, Newman L M, Hur H G, Li S
Department of Biochemistry, Gortner Laboratory, University of Minnesota, St Paul 55108.
Nature. 1994 Apr 14;368(6472):627-9. doi: 10.1038/368627a0.
The decomposition of organic compounds by bacteria has been studied for almost a century, during which time selective enrichment culture has generated microorganisms capable of metabolizing thousands of organic compounds. But attempts to obtain pure cultures of bacteria that can metabolize highly halogenated compounds, a large and important class of pollutants, have been largely unsuccessful. Polyhalogenated compounds are most frequently metabolized by anaerobic bacteria as a result of reductive dehalogenation reactions, the products of which are typically substrates for bacterial oxygenases. Complete metabolism of polyhalogenated compounds therefore necessitates the sequential use of anaerobic and aerobic bacteria. Here we combine seven genes encoding two multi-component oxygenases in a single strain of Pseudomonas which as a result metabolizes polyhalogenated compounds by means of sequential reductive and oxidative reactions to yield non-toxic products. Cytochrome P450cam monooxygenase reduces polyhalogenated compounds, which are bound at the camphor-binding site, under subatmospheric oxygen tensions. We find that these reduction products are oxidizable substrates for toluene dioxygenase. Perhalogenated chlorofluorocarbons also act as substrates for the genetically engineered strain.
细菌对有机化合物的分解已被研究了近一个世纪,在此期间,选择性富集培养产生了能够代谢数千种有机化合物的微生物。但是,试图获得能够代谢高度卤化化合物(一类大量且重要的污染物)的细菌纯培养物的尝试大多没有成功。多卤化化合物最常被厌氧菌通过还原脱卤反应进行代谢,其产物通常是细菌加氧酶的底物。因此,多卤化化合物的完全代谢需要依次使用厌氧菌和好氧菌。在这里,我们将编码两种多组分加氧酶的七个基因组合在一株假单胞菌中,结果该菌株通过依次进行还原和氧化反应来代谢多卤化化合物,从而产生无毒产物。细胞色素P450cam单加氧酶在低于大气压的氧气张力下还原结合在樟脑结合位点的多卤化化合物。我们发现这些还原产物是甲苯双加氧酶的可氧化底物。全卤化氯氟烃也可作为基因工程菌株的底物。