Hinteregger C, Leitner R, Loidl M, Ferschl A, Streichsbier F
Abteilung für Umweltmikrobiologie, Technische Universität Wien, Vienna, Austria.
Appl Microbiol Biotechnol. 1992 May;37(2):252-9. doi: 10.1007/BF00178180.
The phenol-degrading strain Pseudomonas putida EKII was isolated from a soil enrichment culture and utilized phenol up to 10.6 mM (1.0 g.l-1) as the sole source of carbon and energy. Furthermore, cresols, chlorophenols, 3,4-dimethylphenol, and 4-chloro-m-cresol were metabolized as sole substrates by phenol-grown resting cells of strain EKII. Under conditions of cell growth, degradation of these xenobiotics was achieved only in co-metabolism with phenol. Phenol hydroxylase activity was detectable in whole cells but not in cell-free extracts. The specificity of the hydroxylating enzyme was found during transformation of cresols and chlorophenols: ortho- and meta-substituted phenols were degraded via 3-substituted catechols, while degradation of para-substituted phenols proceeded via 4-substituted catechols. In cell-free extracts of phenol-grown cells a high level of catechol 2,3-dioxygenase as well as smaller amounts of 2-hydroxymuconic semialdehyde hydrolyase and catechol 1,2-dioxygenase were detected. The ring-cleaving enzymes were characterized after partial purification by DEAE-cellulose chromatography.
从土壤富集培养物中分离出了能降解苯酚的恶臭假单胞菌EKII菌株,该菌株能将高达10.6 mM(1.0 g·l-1)的苯酚作为唯一碳源和能源加以利用。此外,甲酚、氯酚、3,4-二甲基苯酚和4-氯间甲酚可被菌株EKII以苯酚为生长底物的静息细胞作为唯一底物进行代谢。在细胞生长条件下,这些异生物质只有在与苯酚共代谢时才能实现降解。在全细胞中可检测到苯酚羟化酶活性,但在无细胞提取物中未检测到。在甲酚和氯酚转化过程中发现了羟化酶的特异性:邻位和间位取代的苯酚通过3-取代儿茶酚进行降解,而对位取代的苯酚则通过4-取代儿茶酚进行降解。在以苯酚为生长底物的细胞的无细胞提取物中,检测到高水平的儿茶酚2,3-双加氧酶以及少量的2-羟基粘康酸半醛水解酶和儿茶酚1,2-双加氧酶。通过DEAE-纤维素色谱法部分纯化后,对这些开环酶进行了表征。