Aliphatic nitrilase from a soil-isolated comamonas testosteroni sp.: gene cloning and overexpression, purification and primary structure
| Autor: | Fabienne Soubrier, Anne-Marie Crutz-Le Coq, Dominique Petre, Joel Crouzet, Sophie Levy-Schil, Didier Faucher |
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| Přispěvatelé: | Département Biotechnologie, Rhóne-Poulenc Rorer |
| Rok vydání: | 1995 |
| Předmět: |
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
Chaperonins Molecular Sequence Data Molecular cloning medicine.disease_cause Nitrilase 03 medical and health sciences Bacterial Proteins Aminohydrolases Escherichia coli Genetics medicine [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Amino Acid Sequence Comamonas testosteroni Cloning Molecular Soil Microbiology 030304 developmental biology 0303 health sciences Alcaligenes faecalis Base Sequence Gram-Negative Aerobic Bacteria Sequence Homology Amino Acid biology 030306 microbiology Protein primary structure General Medicine Rhodococcus rhodochrous biology.organism_classification Molecular biology Recombinant Proteins Biochemistry Genes Bacterial Electrophoresis Polyacrylamide Gel Aliphatic nitrilase |
| Zdroj: | Gene Gene, Elsevier, 1995, 161 (1), pp.15-20. ⟨10.1016/0378-1119(95)00242-X⟩ |
| ISSN: | 0378-1119 1879-0038 |
| DOI: | 10.1016/0378-1119(95)00242-x |
| Popis: | An aliphatic nitrilase, active on adiponitrile and cyanovaleric acid, was identified and purified from Comamonas testosteroni sp. (Ct). Oligodeoxyribonucleotide probes were designed from limited amino acid (aa) sequence information and used to clone the corresponding gene, named nitA. High homologies were found at the aa level between Ct nitrilase and the sequences of known nitrilases. Multi-alignment of sequenced nitrilases suggests that Cys163 of Ct plays an essential role in the active site. This hypothesis is strengthened by molecular studies on nitrilases from Alcaligenes faecalis JM3, and Rhodococcus rhodochrous J1 and K22 [Kobayashi et al., Proc. Natl. Acad. Sci. USA 90 (1993) 247–251; J. Biol. Chem. 267 (1992) 20746–20751; Biochemistry 31 (1992) 9000–9007]. Large amounts of an active recombinant enzyme could be produced in Escherichia coli when nitA was overexpressed together with the E. coli groESL genes. |
| Databáze: | OpenAIRE |
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