Amine-synthesizing enzyme N-substituted formamide deformylase: screening, purification, characterization, and gene cloning
| Autor: | Hiroshi Fukatsu, Hiroki Higashibata, Masahiko Goda, Michihiko Kobayashi, Yoshiteru Hashimoto |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Formamide
Molecular Sequence Data Biology Protein Structure Secondary Amidohydrolases chemistry.chemical_compound Benzylamine Formate Amino Acid Sequence Amines Cloning Molecular Peptide sequence Conserved Sequence chemistry.chemical_classification Multidisciplinary Amidohydrolase Molecular mass Sequence Homology Amino Acid Substrate (chemistry) Biological Sciences Chromatography Ion Exchange Recombinant Proteins Molecular Weight Kinetics Enzyme chemistry Biochemistry Sequence Alignment |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 101(38) |
| ISSN: | 0027-8424 |
| Popis: | N-substituted formamide was produced through the hydration of an isonitrile by isonitrile hydratase in the isonitrile metabolism. The former compound was further degraded by a microorganism, strain F164, which was isolated from soil through an acclimatization culture. The N-substituted formamide-degrading microorganism was identified as Arthrobacter pascens . The microbial degradation was found to proceed through an enzymatic reaction, the N-substituted formamide being hydrolyzed to yield the corresponding amine and formate. The enzyme, designated as N-substituted formamide deformylase (NfdA), was purified and characterized. The native enzyme had a molecular mass of ≈61 kDa and consisted of two identical subunits. It stoichiometrically catalyzed the hydrolysis of N -benzylformamide (an N-substituted formamide) to benzylamine and formate. Of all of the N-substituted formamides tested, N -benzylformamide was the most suitable substrate for the enzyme. However, no amides were accepted as substrates. The gene ( nfdA ) encoding this enzyme was also cloned. The deduced amino acid sequence of nfdA exhibited the highest overall sequence identity (28%) with those of regulatory proteins among known proteins. Only the N-terminal region (residues 58–72) of NfdA also showed significant sequence identity (27–73%) to that of each member of the amidohydrolase superfamily, although there was no similarity in the overall sequence except in the above limited region. |
| Databáze: | OpenAIRE |
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