論文 - 詳細
| RRC ID | 5730 |
|---|---|
| 著者 | Furuya T, Nishi T, Shibata D, Suzuki H, Ohta D, Kino K. |
| タイトル | Characterization of orphan monooxygenases by rapid substrate screening using FT-ICR mass spectrometry. |
| ジャーナル | Chem Biol |
| Abstract |
Characterization of orphan enzymes, for which the catalytic functions and actual substrates are still not elucidated, is a significant challenge in the postgenomic era. Here, we describe a general strategy for exploring the catalytic potentials of orphan monooxygenases based on direct infusion analysis by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). Eight cytochromes P450 from Bacillus subtilis were recombinantly expressed in Escherichia coli and subjected to a reconstitution system containing appropriate electron transfer components and many potential substrates. The reaction mixtures were directly analyzed using FT-ICR/MS, and substrates of the putative enzymes were readily identified from the mass spectral data. This allowed identification of previously unreported CYP109B1 substrates and the functional assignment of two putative cytochromes P450, CYP107J1 and CYP134A1. The FT-ICR/MS-based approach can be easily applied to large-scale screening with the aid of the extremely high mass resolution and accuracy. |
| 巻・号 | 15(6) |
| ページ | 563-72 |
| 公開日 | 2008-6-1 |
| DOI | 10.1016/j.chembiol.2008.05.013 |
| PII | S1074-5521(08)00205-6 |
| PMID | 18559267 |
| MeSH | Bacillus subtilis / enzymology Base Sequence Chromatography, High Pressure Liquid Cloning, Molecular Cytochromes / chemistry Cytochromes / genetics Cytochromes / metabolism* DNA Primers Escherichia coli / genetics Fourier Analysis Mass Spectrometry / methods* Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Substrate Specificity |
| IF | 5.915 |
| 引用数 | 27 |
| 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 | 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 | JCM6157 |