This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chapalmadugu, S Articles by Chaudhry, G R Search for Related Content PubMed PubMed Citation Articles by Chapalmadugu, S Articles by Chaudhry, G R

 Previous Article  |  Next Article 

J Bacteriol. 1993 October; 175(20): 6711-6716

research-article

Isolation of a constitutively expressed enzyme for hydrolysis of carbaryl in Pseudomonas aeruginosa.

Department of Biological Sciences, Oakland University, Rochester, Michigan 48309-4401.

ABSTRACT

A hydrolase constitutively expressed in Pseudomonas aeruginosa which converts carbaryl to 1-naphthol was purified 1,767-fold by using a combination of anion-exchange, hydroxylapatite, gel filtration, and hydrophobic interaction chromatography techniques. The presence of Triton X-100 in buffers was necessary for deaggregation and purification of the hydrolase. This is the first membrane-bound hydrolase involved in the hydrolysis of any methylcarbamate pesticide purified from a bacterial source to date. The enzyme exhibited a unique specificity of hydrolyzing only carbaryl (1-naphthyl N-methylcarbamate) but not any other methylcarbamates. The purified enzyme was a monomer with a molecular mass of 65,000 Da. The pH and temperature optima for the enzyme activity were 8.5 and 45 degrees C, respectively. No cofactor requirement for the hydrolase activity could be demonstrated, and none of the divalent cations studied affected the activity of the enzyme. Also, the enzyme activity was not affected by the thiols: dithioerythritol, dithiothreitol, and 2-mercaptoethanol. The Km and Vmax values for carbaryl were 9 microM and 7.9 mumol/min/mg of protein, respectively.

This article has been cited by other articles:

• Swetha, V. P., Phale, P. S. (2005). Metabolism of Carbaryl via 1,2-Dihydroxynaphthalene by Soil Isolates Pseudomonas sp. Strains C4, C5, and C6. Appl. Environ. Microbiol. 71: 5951-5956 [Abstract] [Full Text]  
• Hayatsu, M., Hirano, M., Nagata, T. (1999). Involvement of Two Plasmids in the Degradation of Carbaryl by Arthrobacter sp. Strain RC100. Appl. Environ. Microbiol. 65: 1015-1019 [Abstract] [Full Text]