The mmgA gene from Bacillus subtilis encodes a degradative acetoacetyl-CoA thiolase
| Autor: | Jayme Williams, Jason J. Reddick |
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| Rok vydání: | 2008 |
| Předmět: |
Operon
Bioengineering Bacillus subtilis medicine.disease_cause Applied Microbiology and Biotechnology Chromatography Affinity Substrate Specificity chemistry.chemical_compound Biosynthesis Escherichia coli medicine Acetyl-CoA C-Acetyltransferase Cloning Molecular Gene Chromatography High Pressure Liquid Spores Bacterial biology Thiolase General Medicine biology.organism_classification Molecular biology chemistry Biochemistry Genes Bacterial Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Acetyltransferase Chromatography Gel Acetyl-CoA C-acetyltransferase Electrophoresis Polyacrylamide Gel Biotechnology |
| Zdroj: | Biotechnology Letters. 30:1045-1050 |
| ISSN: | 1573-6776 0141-5492 |
| DOI: | 10.1007/s10529-008-9642-4 |
| Popis: | Early in sporulation, the mother cell compartment of Bacillus subtilis transcribes the mother cell metabolic gene (mmg) operon. The gene mmgA was assigned by other workers using sequence homology as an acetyl-CoA acetyltransferase [E.C. 2.3.1.9]. The gene was overexpressed in Escherichia coli, and the protein was purified by Ni(2+)-affinity chromatography. However, the expected MmgA-catalyzed biosynthesis of acetoacetyl-CoA from acetyl-CoA was undetectable by a standard UV assay, HPLC, and mass spectrometry. These methods indicated a preference for the reverse degradative thiolytic reaction, with a k(cat) of 80 s(-1), and a K(m) of 70 and 50 microM for CoA and acetoacetyl-CoA, respectively. |
| Databáze: | OpenAIRE |
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