Nordlund I, Powlowski J, Shingler V
Unit for Applied Cell and Molecular Biology, University of Umeå, Sweden.
J Bacteriol. 1990 Dec;172(12):6826-33. doi: 10.1128/jb.172.12.6826-6833.1990.
Pseudomonas sp. strain CF600 metabolizes phenol and some of its methylated derivatives via a plasmid-encoded phenol hydroxylase and meta-cleavage pathway. The genes encoding the multicomponent phenol hydroxylase of this strain are located within a 5.5-kb SacI-NruI fragment. We report the nucleotide sequence and the polypeptide products of this 5.5-kb region. A combination of deletion analysis, expression of subfragments in tac expression vectors, and identification of polypeptide products in maxicells was used to demonstrate that the polypeptides observed are produced from the six open reading frames identified in the sequence. Expression of phenol hydroxylase activity in a laboratory Pseudomonas strain allows growth on phenol, owing to expression of this enzyme and the chromosomally encoded ortho-cleavage pathway. This system, in conjunction with six plasmids that each expressed all but one of the polypeptides, was used to demonstrate that all six polypeptides are required for growth on phenol.
假单胞菌属CF600菌株通过质粒编码的苯酚羟化酶和间位裂解途径代谢苯酚及其一些甲基化衍生物。编码该菌株多组分苯酚羟化酶的基因位于一个5.5kb的SacI-NruI片段内。我们报道了这个5.5kb区域的核苷酸序列和多肽产物。通过缺失分析、在tac表达载体中表达亚片段以及在大细胞中鉴定多肽产物相结合的方法，证明了观察到的多肽是由序列中鉴定出的六个开放阅读框产生的。在实验室假单胞菌菌株中表达苯酚羟化酶活性，由于该酶的表达和染色体编码的邻位裂解途径，使得菌株能够在苯酚上生长。该系统与六个质粒（每个质粒表达除一种多肽外的所有多肽）一起，用于证明所有六种多肽都是在苯酚上生长所必需的。
