关键词: 邻苯二酚-2,3-双加氧酶, 菌种筛选, BIOLOG鉴定系统, 阴沟肠杆菌

Abstract: In order to get strains degrading pyrene, which can produce extracellular catechol-2,3-dioxygenase, oil-contaminated soil samples in Karamay of Xinjiang were taken as specimen sources, strains with polycyclic aromatic hydrocarbon were screened using pyrene as the sole carbon and energy source. Testing the extracellular catechol-2,3-dioxygenase the strain produced qualitatively and quantitatively by chromogenic reaction and enzymatic reaction. And the strain was identified according to its morphological characteristics, BIOLOG GENⅢ microplate identification and 16S rRNA gene sequence analysis. Based on single factor and orthogonal experiments, preliminary discussion on the growth characteristics and the best degradation conditions of the strain were carried out. The results showed that strain W39 degrading pyrene could secrete extracellular catechol-2,3-dioxygenase, and after being induced by pyrene, the enzyme producing capacity was increased by 3 times. Strain W39 was identified as Enterobacter cloacae. Its optimum culture conditions was 35℃, pH 7.0, and 50 mg/L pyrene. Therefore, catechol-2,3-dioxygenase is one of the key enzymes in the process of pyrene degradation, and further studies can be conducted on it.

Key words: catechol-2,3-dioxygenase, strain screening, BIOLOG identification system, Enterobacter cloacae