Kivisaar M A, Habicht J K, Heinaru A L
Laboratory of Plasmid Biology, Estonian Biocenter, Tartu, USSR.
J Bacteriol. 1989 Sep;171(9):5111-6. doi: 10.1128/jb.171.9.5111-5116.1989.
The utilization of phenol, m-toluate, and salicylate (Phe+, mTol+, and Sal+ characters, respectively) in Pseudomonas sp. strain EST1001 is determined by the coordinated expression of genes placed in different plasmids, i.e., by a multiplasmid system. The natural multiplasmid strain EST1001 is phenotypically unstable. In its Phe-, mTol-, and Sal- segregants, the plasmid DNA underwent structural rearrangements without a marked loss of plasmid DNA, and the majority of segregants gave revertants. The genes specifying the degradation of phenol and m-toluate were transferable to P. putida PaW340, and in this strain a new multiplasmid system with definite structural changes was formed. The 17-kilobase transposable element, a part of the TOL plasmid pWWO present in the chromosome of PaW340, was inserted into the plasmid DNA in transconjugants. In addition, transconjugant EST1020 shared pWWO-like structures. Enzyme assays demonstrated that ortho-fission reactions were used by bacteria that grew on phenol, whereas m-toluate was catabolized by a meta-fission reaction. Salicylate was a functional inducer of the enzymes of both pathways. The expression of silent metabolic pathways of phenol or m-toluate degradation has been observed in EST1001 Phe- mTol+ and Phe+ mTol- transconjugants. The switchover of phenol degradation from the ortho- to the meta-pathway in EST1033 also showed the flexibility of genetic material in EST1001 transconjugants.
假单胞菌属菌株EST1001中对苯酚、间甲苯酸盐和水杨酸盐(分别为Phe +、mTol +和Sal +特性)的利用是由位于不同质粒中的基因的协调表达决定的,即由多质粒系统决定。天然多质粒菌株EST1001在表型上不稳定。在其Phe -、mTol -和Sal -分离株中,质粒DNA发生了结构重排,但质粒DNA没有明显损失,并且大多数分离株产生了回复突变体。指定苯酚和间甲苯酸盐降解的基因可转移到恶臭假单胞菌PaW340中,并且在该菌株中形成了具有确定结构变化的新多质粒系统。17千碱基的转座元件,即存在于PaW340染色体中的TOL质粒pWWO的一部分,在转接合子中插入到质粒DNA中。此外,转接合子EST1020具有pWWO样结构。酶分析表明,在苯酚上生长的细菌使用邻位裂解反应,而间甲苯酸盐通过间位裂解反应进行分解代谢。水杨酸盐是这两种途径中酶的功能性诱导剂。在EST1001 Phe - mTol +和Phe + mTol -转接合子中观察到了苯酚或间甲苯酸盐降解的沉默代谢途径的表达。EST1033中苯酚降解从邻位途径到间位途径的转换也显示了EST1001转接合子中遗传物质的灵活性。