論文 - 詳細
| RRC ID | 29290 |
|---|---|
| 著者 | Fedorov DN, Ekimova GA, Doronina NV, Trotsenko YA. |
| タイトル | 1-Aminocyclopropane-1-carboxylate (ACC) deaminases from Methylobacterium radiotolerans and Methylobacterium nodulans with higher specificity for ACC. |
| ジャーナル | FEMS Microbiol Lett |
| Abstract |
The 1-aminocyclopropane-1-carboxylate (ACC) deaminases (EC 3.4.99.7), the key enzymes of degradation of the precursor of the phytohormone ethylene, have not been well studied despite their great importance for plant-bacterial interactions. Using blast, the open reading frames encoding ACC deaminases were found in the genomes of epiphytic methylotroph Methylobacterium radiotolerans JCM2831 and nodule-forming endosymbiont Methylobacterium nodulans ORS2060. These genes were named acdS and cloned; recombinant proteins were expressed and purified from Escherichia coli. The enzyme from M. nodulans displayed the highest substrate specificity among all of the characterized ACC deaminases (Km 0.80 ± 0.04 mM), whereas the enzyme from M. radiotolerans had Km 1.8 ± 0.3 mM. The kcat values were 111.8 ± 0.2 and 65.8 ± 2.8 min(-1) for the enzymes of M. nodulans and M. radiotolerans, respectively. Both enzymes are homotetramers with a molecular mass of 144 kDa, as was demonstrated by size exclusion chromatography and native PAGE. The purified enzymes displayed the maximum activity at 45-50 °C and pH 8.0. Thus, the priority data have been obtained, extending the knowledge of biochemical properties of bacterial ACC deaminases. |
| 巻・号 | 343(1) |
| ページ | 70-6 |
| 公開日 | 2013-6-1 |
| DOI | 10.1111/1574-6968.12133 |
| PMID | 23517598 |
| MeSH | Carbon-Carbon Lyases / chemistry Carbon-Carbon Lyases / genetics Carbon-Carbon Lyases / isolation & purification Carbon-Carbon Lyases / metabolism* Chromatography, Gel Cloning, Molecular Computational Biology Electrophoresis, Polyacrylamide Gel Enzyme Stability Escherichia coli / genetics Gene Expression Hydrogen-Ion Concentration Kinetics Methylobacterium / enzymology* Methylobacterium / genetics Molecular Weight Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Homology Substrate Specificity Temperature |
| IF | 1.987 |
| 引用数 | 15 |
| WOS 分野 | MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| 一般微生物 | JCM 2831 |