Lactobacillus reuteri 2′-Deoxyribosyltransferase, a Novel Biocatalyst for Tailoring of Nucleosides
| Autor: | Miguel Arroyo, Jesús Fernández-Lucas, Isabel de la Mata, José V. Sinisterra, Carmen Acebal |
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| Rok vydání: | 2010 |
| Předmět: |
Limosilactobacillus reuteri
Circular dichroism Hot Temperature Gene Expression medicine.disease_cause Applied Microbiology and Biotechnology Protein Structure Secondary Bacterial Proteins Enzyme Stability Escherichia coli medicine Pentosyltransferases Cloning Molecular Enzymology and Protein Engineering Arabinosyltransferase activity Protein secondary structure chemistry.chemical_classification Ecology biology Circular Dichroism Nucleosides Hydrogen-Ion Concentration biology.organism_classification Enzymes Lactobacillus reuteri Molecular Weight Kinetics Enzyme chemistry Biochemistry Sedimentation equilibrium Protein Multimerization Nucleoside Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 76:1462-1470 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.01685-09 |
| Popis: | A novel type II nucleoside 2′-deoxyribosyltransferase from Lactobacillus reuteri ( Lr NDT) has been cloned and overexpressed in Escherichia coli . The recombinant Lr NDT has been structural and functionally characterized. Sedimentation equilibrium analysis revealed a homohexameric molecule of 114 kDa. Circular dichroism studies have showed a secondary structure containing 55% α-helix, 10% β-strand, 16% β-sheet, and 19% random coil. Lr NDT was thermostable with a melting temperature ( T m ) of 64°C determined by fluorescence, circular dichroism, and differential scanning calorimetric studies. The enzyme showed high activity in a broad pH range (4.6 to 7.9) and was also very stable between pH 4 and 7.9. The optimal temperature for activity was 40°C. The recombinant Lr NDT was able to synthesize natural and nonnatural nucleoside analogues, improving activities described in the literature, and remarkably, exhibited unexpected new arabinosyltransferase activity, which had not been described so far in this kind of enzyme. Furthermore, synthesis of new arabinonucleosides and 2′-fluorodeoxyribonucleosides was carried out. |
| Databáze: | OpenAIRE |
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