Over-expression in Escherichia coli of a thermally stable and regio-selective nitrile hydratase from Comamonas testosteroni 5-MGAM-4D
| Autor: | Eugenia Costa Hann, Kelly L. Petrillo, Mark S. Payne, John E. Gavagan, Frederick B. Cooling, Robert DiCosimo, Arie Ben-Bassat, Shijun Wu |
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| Rok vydání: | 2005 |
| Předmět: |
DNA
Bacterial Nitrile Molecular Sequence Data Gene Expression Biology medicine.disease_cause Applied Microbiology and Biotechnology Amidohydrolases Substrate Specificity Amidase Open Reading Frames chemistry.chemical_compound Bacterial Proteins Nitrile hydratase Enzyme Stability Escherichia coli medicine Amino Acid Sequence Comamonas testosteroni Cloning Molecular Hydro-Lyases chemistry.chemical_classification Base Sequence Temperature Sequence Analysis DNA General Medicine equipment and supplies biology.organism_classification Enterobacteriaceae Recombinant Proteins Protein Subunits Open reading frame Enzyme Biochemistry chemistry Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology. 67:664-670 |
| ISSN: | 1432-0614 0175-7598 |
| Popis: | The genes encoding a thermally stable and regio-selective nitrile hydratase (NHase) and an amidase from Comamonas testosteroni 5-MGAM-4D have been cloned and sequenced, and active NHase has been over-produced in Escherichia coli. Maximal activity requires co-expression of a small open reading frame immediately downstream from the NHase beta subunit gene. Compared to the native organism, the E. coli biocatalyst has nearly threefold more NHase activity on a dry cell weight basis, and this activity is significantly more thermally stable. In addition, this biocatalyst converts a wide spectrum of nitrile substrates to the corresponding amides. Such versatility and robustness are desirable attributes of a biocatalyst intended for use in commercial applications. |
| Databáze: | OpenAIRE |
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