| Autor: |
Krieg, L., Slusarczyk, H., Verseck, S., Kula, M.-R. |
| Předmět: |
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| Zdroj: |
Applied Microbiology & Biotechnology; Mar2005, Vol. 66 Issue 5, p542-550, 9p, 5 Diagrams, 1 Chart |
| Abstrakt: |
The gene for the newly describedd-amidase fromVariovorax paradoxus(Krieg et al. 2002) was cloned and functionally expressed inEscherichia coli. Since native enzyme was available in minute amounts only, we determined the N-terminal sequence of the enzyme and utilized the Universal GenomeWalker Approach to make use of the common internal sequence of the amidase signature family. The high GC content of the gene made it necessary to employ an appropriate DNA polymerase in the amplification reactions. Thus, the sequence of the complete gene and the flanking regions was established. In independent experiments, the gene was then amplified from genomic DNA ofV. paradoxus, expressed inE. coli, and characterized. The recombinant enzyme has a specific activity of 1.7 units/mg with racemictert-leucine amide as substrate and is a homodimer of 49.6-kDa monomers. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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