Shingler V, Franklin F C, Tsuda M, Holroyd D, Bagdasarian M
Unit for Applied Cell and Molecular Biology, University of Umeå, Sweden.
J Gen Microbiol. 1989 May;135(5):1083-92. doi: 10.1099/00221287-135-5-1083.
Pseudomonas strain CF600 is able to utilize phenol and 3,4-dimethylphenol as sole carbon and energy source. We demonstrate that growth on these substrates is by virtue of plasmid-encoded phenol hydroxylase and a meta-cleavage pathway. Screening of a genomic bank, with DNA from the previously cloned catechol 2,3-dioxygenase gene of the TOL plasmid pWW0, was used in the identification of a clone which could complement a phenol-hydroxylase-deficient transposon insertion mutant. Deletion mapping and polypeptide production analysis identified a 1.2 kb region of DNA encoding a 39.5 kDa polypeptide which mediated this complementation. Enzyme activities and growth properties of Pseudomonas strains harbouring this fragment on a broad-host-range expression vector indicate that phenol hydroxylase is a multicomponent enzyme containing the 39.5 kDa polypeptide as one component.
假单胞菌CF600菌株能够利用苯酚和3,4 - 二甲基苯酚作为唯一的碳源和能源。我们证明，在这些底物上的生长是借助于质粒编码的苯酚羟化酶和间位裂解途径。用来自先前克隆的TOL质粒pWW0的儿茶酚2,3 - 双加氧酶基因的DNA筛选基因组文库，用于鉴定一个可以互补苯酚羟化酶缺陷型转座子插入突变体的克隆。缺失定位和多肽产生分析确定了一个1.2 kb的DNA区域，该区域编码一种39.5 kDa的多肽，介导了这种互补作用。携带该片段的假单胞菌菌株在广宿主范围表达载体上的酶活性和生长特性表明，苯酚羟化酶是一种多组分酶，其中39.5 kDa的多肽是其一个组分。
