Sequence of the gene ( pheA) encoding phenol monooxygenase from Pseudomonas sp. EST1001: expression in Escherichia coli and Pseudomonas putida

• Published in: Gene Vol. 102; no. 1; pp. 13 - 18
• Main Authors: Nurk, Allan, Kasak, Lagle, Kivisaar, Maia
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 15.06.1991; Amsterdam Elsevier; New York, NY
• Subjects: Amino Acid Sequence; amino acid sequence comparison; Base Sequence; Biological and medical sciences; Cloning, Molecular; DNA Transposable Elements - genetics; Escherichia coli - metabolism; flavoprotein aromatic monooxygenases; Flavoproteins - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression - physiology; genes; Genes. Genome; Mixed Function Oxygenases - genetics; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; nucleotide sequence; pheA gene; phenol 2-monooxygenase; phenol degradation; Plasmids - genetics; predictions; Pseudomonas; Pseudomonas - enzymology; Pseudomonas - genetics; Recombinant DNA; Sequence Homology, Nucleic Acid; aa; ORF; RBS; SDS; 2,4-D; nt; 2,4-DCPH; PAGE; Recombinant DNA; flavoprotein aromatic monooxygenases; pheA and pheB; bp; Ap; P; C120; PMO; PolIk; LB; kb; amino acid sequence comparison; phenol degradation; Cb; Pseudomonadales; Nucleotide sequence; Enzyme; Homology; Phenol 2-monooxygenase; Gene expression; Pseudomonas; Plasmid; Enzymatic activity; DNA; Bacteria; Pseudomonadaceae; Molecular cloning
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(91)90531-F

The plasmid pEST1412 contains the genes, pheA and pheB, encoding phenol monooxygenase (PMO) and catechol 1,2-dioxygenase (C120), respectively. These were originally cloned from the plasmid DNA of Pseudomonas sp. EST1001 [Kivisaar et al., Plasmid 24 (1990) 25–36]. Although pheA and pheB are cotranscribed using the promoter sequences derived from Tn4652 and the level of expression of C120 activities from pEST1412 was equal both in Escherichia coli and in Pseudomonas putida, the level of PMO activity measured in the cell-free extracts of E. coli was lower than that in P. putida. The nucleotide sequence of the 2.0-kb PstI- HindIII fragment of pEST1412 carrying pheA was determined. A 1821-bp ORF was found in this DNA. The structural gene ( tfdB) encoding 2,4-dichlorophenol hydroxylase from pJP4 has been sequenced [Perkins et al., J. Bacteriol. 172 (1990) 2351–2359]. Comparison of the deduced amino acid sequences of tfdB and pheA revealed highly conserved regions in the protein products of these genes.