論文 - 詳細
| RRC ID | 78969 |
|---|---|
| 著者 | Shunsuke Kato, Koki Takeuchi, Motonao Iwaki, Kentaro Miyazaki, Kohsuke Honda, Takashi Hayashi |
| タイトル | Chitin‐ and Streptavidin‐Mediated Affinity Purification Systems: A Screening Platform for Enzyme Discovery |
| ジャーナル | Angewandte Chemie International Edition |
| Abstract |
Affinity purification of recombinant proteins is an essential technique in biotechnology. However, current affinity purification methods are very cost‐intensive, and this imposes limits on versatile use of affinity purification for obtaining purified proteins for a variety of applications. To overcome this problem, we developed a new affinity purification system which we call CSAP (chitin‐ and streptavidin‐mediated affinity purification) for low‐cost purification of Strep‐tag II fusion proteins. The CSAP system is designed to utilize commercially available chitin powder as a chromatography matrix, thereby significantly improving the cost‐efficiency of protein affinity purification. We investigated the CSAP system for protein screening in 96‐well format as a demonstration. Through the screening of 96 types of purified hemoproteins, several proteins capable of the catalytic diastereodivergent synthesis of cyclopropanes were identified as candidates for an abiotic carbene transfer reaction. |
| 巻・号 | 62 |
| 公開日 | 2023-8-1 |
| DOI | 10.1002/anie.202303764 |
| PMID | 37278513 |
| MeSH | Chitin* / chemistry Chromatography, Affinity / methods Escherichia coli* / metabolism Recombinant Fusion Proteins / chemistry Recombinant Proteins / chemistry Streptavidin / chemistry |
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 11 |
| 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) | |