The final acylation step in aromatic dithiolopyrrolone biosyntheses: identification and characterization of the first bacterium N-benzoyltransferase from Saccharothrix algeriensis NRRL B-24137
| Autor: | Safwan Saker, Florence Mathieu, Stéphanie Chacar |
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| Přispěvatelé: | Centre National de la Recherche Scientifique - CNRS (FRANCE), Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE), Institut National de la Recherche Agronomique - INRA (FRANCE), Université Toulouse III - Paul Sabatier - UT3 (FRANCE), Université de Lorraine (FRANCE), Laboratoire de génie chimique [ancien site de Basso-Cambo] (LGC), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Dynamique des Génomes et Adaptation Microbienne (DynAMic), Université de Lorraine (UL)-Institut National de la Recherche Agronomique (INRA), Institut National Polytechnique de Toulouse - INPT (FRANCE), Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL) |
| Rok vydání: | 2014 |
| Předmět: |
Stereochemistry
Acylation Benzoyl-holothin Bioengineering Saccharothrix algeriensis medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry Substrate Specificity chemistry.chemical_compound [CHIM.GENI]Chemical Sciences/Chemical engineering Benzoyl-pyrrothine Biosynthesis Bacterial Proteins N-benzoyltransferase medicine Génie chimique [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering Génie des procédés Escherichia coli Pyrrothine chemistry.chemical_classification biology Chemistry Temperature Hydrogen-Ion Concentration biology.organism_classification Pyrrolidinones Recombinant Proteins Anti-Bacterial Agents Actinobacteria Kinetics Enzyme Acyltransferases Genes Bacterial Acetyltransferase bacteria Holothin Bacteria Acyl group Biotechnology |
| Zdroj: | Enzyme and Microbial Technology Enzyme and Microbial Technology, Elsevier, 2015, 72, pp.35-41. ⟨10.1016/j.enzmictec.2015.02.005⟩ |
| ISSN: | 1879-0909 0141-0229 |
| Popis: | International audience; The last step in the biosynthesis of dithiolopyrrolone antibiotics was thought to involve the transfer of acyl group from acyl-CoA to pyrrothine/holothin core. In Saccharothrix algeriensis NRRL B-24137, two acyltransferases, an acetyltransferase and a benzoyltransferase were proposed to catalyze this step. We have previously identified, in Sa. algeriensis genome, two open read frames, actA and actB patiently encoded these enzymes. This study focuses primarily on the characterization of the protein encoded by actA. After cloning and expressing of actA in Escherichia coli BL21, the recombinant protein encoded by actA was purified. Selectivity of ActA for pyrrothine/holothin as substrate and different acyl-CoA as co-substrate was evaluated using two acyls-groups, linear and aromatic. The enzyme was shown to prefer aromatic groups over linear groups as donor group; further neither product nor transfer was observed for linear groups. Therefore ActA has been determined to be a pyrrothine/holothin N-benzoyltransferase which can either pyrrothine (Km of 72 μM) or holothin (Km of 129.5 μM) as substrates and benzoyl-CoA (Km of 348.65 and 395.28 μM) as co-substrates for pyrrothine and holothin, respectively. The optimum pH and temperature has been shown to be 8, 40 °C, respectively. ActA is the first enzyme characterized as N-benzoyltransferase in bacteria. |
| Databáze: | OpenAIRE |
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