Purification and biochemical characterization of the carbamate hydrolase from Pseudomonas sp. 50432
| Autor: | A. Mateen, G. Rasul Chaudhry, Sheikh Riazuddin, M. Bloda, B. Kaskar |
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| Rok vydání: | 2002 |
| Předmět: |
Biomedical Engineering
Bioengineering Applied Microbiology and Biotechnology Sensitivity and Specificity chemistry.chemical_compound Hydrolysis Carbofuran Carbaryl Pseudomonas Drug Discovery Enzyme Stability Mercaptoethanol chemistry.chemical_classification Chromatography biology Molecular mass Process Chemistry and Technology Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Enzyme assay Molecular Weight Enzyme chemistry Biochemistry Models Chemical biology.protein Molecular Medicine Carbamates Carboxylic Ester Hydrolases Biotechnology Pseudomonadaceae |
| Zdroj: | Biotechnology and applied biochemistry. 36(1) |
| ISSN: | 0885-4513 |
| Popis: | A soluble carbamate hydrolase that had a wide specificity was purified 2032-fold from Pseudomonas sp. 50432. This was achieved using a combination of anion-exchange, gel-filtration and hydrophobic-interaction- chromatography techniques. Carbamate hydrolase cleaved the ester linkage of the N-methylcarbamates. The native enzyme was a monomer with a molecular mass of 88 kDa. The optimum pH and temperature of the enzyme activity were 8.5 and 37 degrees C respectively. The tested cations or EDTA did not affect the enzyme activity. However, 2-mercaptoethanol reversibly inhibited the enzyme activity. The enzyme showed the K(m) values of 16 and 12 microM for carbofuran and carbaryl respectively. The purified enzyme did not hydrolyse o-nitrophenyl dimethylcarbamate but hydrolysed several N-methylcarbamates and 1-naphthyl acetate. |
| Databáze: | OpenAIRE |
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