論文 - 詳細
| RRC ID | 59084 |
|---|---|
| 著者 | Goto H, Kanai Y, Yotsui A, Shimokihara S, Shitara S, Oyobiki R, Fujiwara K, Watanabe T, Einaga Y, Matsumoto Y, Miki N, Doi N. |
| タイトル | Microfluidic screening system based on boron-doped diamond electrodes and dielectrophoretic sorting for directed evolution of NAD(P)-dependent oxidoreductases. |
| ジャーナル | Lab Chip |
| Abstract |
We report the development of a micro total analysis system (μTAS) based on electrochemical measurements and dielectrophoretic sorting for screening of NAD(P)-dependent oxidoreductases. In this system, the activity of enzymes immobilized on microbeads, together with their encoding DNA, can be measured with a boron-doped diamond (BDD) electrode in each compartment (∼30 nL) of the microfluidic system. The 30 nL droplets containing microbead-displayed genes of enzymes with higher activity can then be recovered by dielectrophoretic sorting. Previously, we developed the NAD(P)H-measuring device containing the BDD electrode for high-throughput measurement of the activity of NAD(P)-dependent oxidoreductases. In this study, we fabricated an encapsulating device and a droplet-sorting device for nanoliter-size droplets, for the first time, and then combined these three devices to construct a μTAS for directed evolution of NAD(P)-dependent oxidoreductases. We confirmed that this system works by proof-of-principle experiments and successfully applied this system for screening of randomized libraries of NAD-dependent dehydrogenases. |
| 巻・号 | 20(4) |
| ページ | 852-861 |
| 公開日 | 2020-2-21 |
| DOI | 10.1039/c9lc01263j |
| PMID | 31984406 |
| MeSH | Boron* Diamond* Electrodes Microfluidics NAD Oxidoreductases / genetics |
| IF | 6.914 |
| 引用数 | 0 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 3 |
| 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) | iVEC strain |