Cloning, nucleotide sequence and expression of a new --carbamoylase gene from DSM 3747 in
| Autor: | Anja Wiese, Christoph Syldatk, Josef Altenbuchner, Ralf Mattes, Markus Pietzsch, Burkhard Wilms |
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| Rok vydání: | 1999 |
| Předmět: |
chemistry.chemical_classification
Amidohydrolase Nucleic acid sequence Bioengineering General Medicine Molecular cloning Biology medicine.disease_cause Applied Microbiology and Biotechnology Amino acid Enzyme Isoelectric point Biochemistry chemistry medicine Escherichia coli Peptide sequence Biotechnology |
| Zdroj: | Journal of Biotechnology. 68:101-113 |
| ISSN: | 0168-1656 |
| Popis: | An l -N-carbamoyl amino acid amidohydrolase ( l -N-carbamoylase) from Arthrobacter aurescens DSM 3747 was cloned in E. coli and the nucleotide sequence was determined. After expression of the gene in E. coli the enzyme was purified to homogeneity and characterized. The enzyme was shown to be strictly l -specific and exhibited the highest activity in the hydrolysis of β-aryl substituted Nα-carbamoyl-alanines as e.g. N-carbamoyl-tryptophan. Carbamoyl derivatives of β-alanine and charged aliphatic amino acids were not accepted as substrates. The N-carbamoylase of A. aurescens DSM 3747 differs from all known enzymes with respect to its substrate specificity although amino acid sequence identity scores of 35–38% to other N-carbamoylases have been detected. The enzyme consists of two subunits of 44.000 Da, and has an isoelectric point of 4.3. The optima of temperature and pH were determined to be 50°C and pH 8.5 respectively. At 37°C the enzyme was completely stable for several days. |
| Databáze: | OpenAIRE |
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