論文 - 詳細
| RRC ID | 35617 |
|---|---|
| 著者 | Zhou S, Mohan Raj S, Ashok S, Edwardraja S, Lee SG, Park S. |
| タイトル | Cloning, expression and characterization of 3-hydroxyisobutyrate dehydrogenase from Pseudomonas denitrificans ATCC 13867. |
| ジャーナル | PLoS One |
| Abstract |
The gene encoding an NAD(+)-dependent, 3-hydroxyisobutyrate dehydrogenase (3HIBDH-IV) from Pseudomonas denitrificans ATCC 13867 was cloned and expressed in Escherichia coli BL 21 (DE3) and characterized to understand its physiological relevance in the degradation of 3-hydroxypropionic acid (3-HP). The deduced amino acid sequence showed high similarity to other 3-hydroxyisobutyrate dehydrogenase isozymes (3HIBDHs) of P. denitrificans ATCC 13867. A comparison of 3HIBDH-IV with its relevant enzymes along with molecular docking studies suggested that Lys171, Asn175 and Gly123 are important for its catalytic function on 3-hydroxyacids. The recombinant 3HIBDH-IV was purified to homogeneity utilizing a Ni-NTA-HP resin column in high yield. 3HIBDH-IV was very specific to (S)-3-hydroxyisobutyrate, but also catalyzed the oxidation of 3-HP to malonate semialdehyde. The specific activity and half-saturation constant (K m) for 3-HP at 30°C and pH 9.0 were determined to be 17 U/mg protein and 1.0 mM, respectively. Heavy metals, such as Ag(+) and Hg(2+), completely inhibited the 3HIBDH-IV activity, whereas dithiothreitol, 2-mercaptoethanol and ethylenediaminetetraacetic acid increased its activity 1.5-1.8-fold. This paper reports the characteristics of 3HIBDH-IV as well as its probable role in 3-HP degradation. |
| 巻・号 | 8(5) |
| ページ | e62666 |
| 公開日 | 2013-1-1 |
| DOI | 10.1371/journal.pone.0062666 |
| PII | PONE-D-12-33376 |
| PMID | 23658760 |
| PMC | PMC3642240 |
| MeSH | Alcohol Oxidoreductases / chemistry Alcohol Oxidoreductases / genetics Alcohol Oxidoreductases / metabolism* Amino Acid Sequence Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* Catalytic Domain Chelating Agents / chemistry Cloning, Molecular Edetic Acid / chemistry Enzyme Stability Hydrogen-Ion Concentration Kinetics Lactic Acid / analogs & derivatives* Lactic Acid / metabolism Molecular Docking Simulation Molecular Sequence Data Protein Structure, Secondary Pseudomonas / enzymology* Sequence Analysis, DNA Sequence Homology, Amino Acid Structural Homology, Protein Transition Elements / chemistry |
| IF | 2.74 |
| 引用数 | 9 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| 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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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 9 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 原核生物(大腸菌) | |