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MOLECULAR CLONING, CHARACTERIZATION, AND REGULATION OF A PSEUDOMONAS PICKETTII PK01 GENE IN ENCODING PHENOL HYDROXYLASE AND EXPRESSION OF THE GENE IN PSEUDOMONAS AERUGINOSA PA01C
Citation:
Kukor, J. AND R. Olsen. MOLECULAR CLONING, CHARACTERIZATION, AND REGULATION OF A PSEUDOMONAS PICKETTII PK01 GENE IN ENCODING PHENOL HYDROXYLASE AND EXPRESSION OF THE GENE IN PSEUDOMONAS AERUGINOSA PA01C. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-90/379 (NTIS PB91163923), 1990.
Description:
A 26 kilobase BamHI restriction endonuclease DNA fragment has been cloned from Pseudomonas pickettii PK01, a strain isolated from a soil microcosm that had been amended with benzene, toluene, and xylene. This DNA fragment, cloned into vector plasmid pR0l727 and designated pR0l957, allowed P. aeruginosa PA01c to grow on phenol as sole source of carbon. hysical and functional restriction endonuclease maps have ben derived for the cloned DNA fragment. wo DNA fragments carried in trans and derived from subclones of pR0l957 show phenol hydroxylase activity in cell-free extracts of P. aeruginosa. eletion and subcloning analyses of these fragments indicated that the gene encoding phenol hydroxylase is positively regulated. henol and m-cresol were shown to be inducers of the enzyme. -Cresol and p-cresol did not induce enzymatic activity, but could be metabolized by cells that had been previously exposed to phenol or m-cresol, moreover the enzyme exhibited a rather broad substrate specificity and was sensitive to thiol-inhibiting reagents. ovel polypeptide with an estimated molecular mass of 80,000 daltons was detected in extracts of phenol-induced cells of P. aeruginosa carrying plasmid pR0l959.
