論文 - 詳細
| RRC ID | 15467 |
|---|---|
| 著者 | Kakugawa S, Fushinobu S, Wakagi T, Shoun H. |
| タイトル | Characterization of a thermostable carboxylesterase from the hyperthermophilic bacterium Thermotoga maritima. |
| ジャーナル | Appl Microbiol Biotechnol |
| Abstract |
The gene encoding carboxylesterase from the hyperthermophilic bacterium Thermotoga maritima (tm0053) was cloned. The recombinant protein (EST53) was overexpressed in Escherichia coli without its NH(2)-terminal hydrophobic region, and with a C-terminal hexahistidine sequence. The enzyme was purified to homogeneity by heat treatment, followed by Ni(2+) affinity chromatography, and then characterized. Among the p-nitrophenyl esters tested, the best substrate was p-nitrophenyl decanoate with K (m) and k (cat) values of 3.1 muM and 10.8 s(-1), respectively, at 60 degrees C and pH 7.5. The addition of O,O'-bis(2-aminoethyl)ethyleneglycol-N,N,N',N'-tetraacetic acid decreased the esterase activity, indicating that EST53 is dependent on the presence of Ca(2+) ion. In addition, its activity was inhibited by the addition of phenylmethylsulfonyl fluoride and diethyl pyrocarbonate, indicating that it contains serine and histidine residues, which play key roles in the catalytic mechanism. EST53 shows a relatively high degree of similarity to Burkholderia lipases that belong to family I.2 of the lipolytic enzymes, whereas the local sequence around the pentapeptide of EST53 is most similar to those of Bacillus lipases belonging to family I.4. |
| 巻・号 | 74(3) |
| ページ | 585-91 |
| 公開日 | 2007-3-1 |
| DOI | 10.1007/s00253-006-0687-9 |
| PMID | 17106678 |
| MeSH | Amino Acid Sequence Bacillus / genetics Burkholderia / genetics Calcium / pharmacology Carboxylesterase / chemistry Carboxylesterase / genetics Carboxylesterase / isolation & purification Carboxylesterase / metabolism* Chemical Fractionation Chromatography, Affinity Cloning, Molecular Diethyl Pyrocarbonate / pharmacology Enzyme Activators / pharmacology Enzyme Inhibitors / pharmacology Enzyme Stability Escherichia coli Hot Temperature Lipase / genetics Molecular Sequence Data Phenylmethylsulfonyl Fluoride / pharmacology Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Homology, Amino Acid Substrate Specificity Thermotoga maritima / enzymology* Thermotoga maritima / genetics |
| IF | 3.53 |
| 引用数 | 33 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED 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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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 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 | |
| 一般微生物 | JCM 10099 |