A new variant activator involved in the degradation of phenolic compounds from a strain of Pseudomonas putida.
A new variant type of regulatory activator and relevant promoters (designated capR, Pr and Po) involved in the metabolism of phenolic compounds were cloned from Pseudomonas putida KCTC1452 by using PCR. The deduced amino acid sequence of CapR revealed a difference in nine amino acids from the effector binding domain of DmpR. To measure effector specificity, plasmids were constructed in such a way that the expression of luc gene for firefly luciferase or lacZ for beta-galactosidase as a reporter was under the control of capR. When Escherichia coli transformed with the plasmids was exposed to phenol, dramatic increases in the activity of luciferase or beta-galactosidase were observed in a range of 0.01-1 mM. Among various phenolic compounds tested, other effective compounds included catechol, 2-methylphenol, 3-methylphenol, 4-methylphenol, 2-chlorophenol, 4-chlorophenol, 2-nitrophenol, resorcinol, and 2, 5-dimethylphenol. The results indicate that CapR has effector specificity different from other related activators, CatR and DmpR. Waste water and soil potentially containing phenolic compounds were also tested by this system and the results were compared with chemical and GC data. The present results indicate that the biosensor consisting of capR and the promoters may be utilized for the development of a phenolic compounds-specific biosensor in monitoring the environmental pollutant.
Park
Seun Mi
SM
Department of Molecular Biology, College of Natural Sciences, Pusan National University, Busan 609-735, South Korea.
Park
Hoo Hwi
HH
Lim
Woon Ki
WK
Shin
Hae Ja
HJ
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D015374:Biosensing Techniques / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism*; D051922:PII Nitrogen Regulatory Proteins; D010636:Phenols / Q000032:analysis* / Q000493:pharmacokinetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015202:Protein Engineering / Q000379:methods; D016958:Pseudomonas putida / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors; D015533:Transcriptional Activation / Q000235:genetics; D001616:beta-Galactosidase / Q000378:metabolism*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C080133:DmpR protein, Pseudomonas; D051922:PII Nitrogen Regulatory Proteins; D010636:Phenols; D015534:Trans-Activators; D014157:Transcription Factors; D008156:Luciferases; D001616:beta-Galactosidase