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-Structure paper
| タイトル | Structure-guided engineering of a Rieske-type aromatic dioxygenase for enhanced consumption of 3-phenylpropionic acid in Escherichia coli. |
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| ジャーナル・号・ページ | J Hazard Mater, Vol. 491, Page 137954, Year 2025 |
| 掲載日 | 2025年7月5日 |
著者 | Wenxue Jiang / Miao Wu / Zhou Gong / Linhua Han / Xiaoqi Cheng / Xiaoqin Tang / Xiaolong Yu / Xu Dong / Yibin Cheng / Lixin Ma / Qiong Xing / ![]() |
| PubMed 要旨 | Industrial derived aromatic hydrocarbons are persistent environmental pollutants due to their chemical stability, posing both ecological and health risks. Rieske-type aromatic dioxygenases (RDOs), ...Industrial derived aromatic hydrocarbons are persistent environmental pollutants due to their chemical stability, posing both ecological and health risks. Rieske-type aromatic dioxygenases (RDOs), known for their role in dihydroxylation of aromatic rings, play a pivotal role in microbial consumption and degradation of such compounds. While the industrial application of these enzymes has been impeded by their instability and low biodegradation rate. In this study, we focused on optimization and application of the Rieske-type dioxygenase HcaEF from Escherichia coli (E. coli) K-12, which initializes the degradation of 3-phenylpropionic acid (3-PP) and cinnamic acid (CI). Using cryo-electron microscopy (cryo-EM), we determined the high-resolution structures of the apo-form and 3-PP bound form of HcaEF, revealing key insights into substrate specificity and thermal stability. Leveraging these structural insights, we engineered a Q73I variant of HcaEF. Upon introduction of this mutation, the turnover rate increased from 29.6 % to 43.8 %, showing ∼50 % improvement. Overexpression of this variant in E. coli K-12 significantly enhanced the strain's ability to utilize 3-PP, demonstrating the potential for microbial engineering in environmental bioremediation and industrial applications. Our findings not only deepen the understanding of substrate recognition in RDOs, but also pave the way for developing high-efficiency enzymes for aromatic compound bio-utilization. |
リンク | J Hazard Mater / PubMed:40120277 |
| 手法 | EM (単粒子) |
| 解像度 | 3.1 Å |
| 構造データ | EMDB-62756, PDB-9l1w: |
| 化合物 | ![]() ChemComp-FE2: ![]() ChemComp-FES: ![]() ChemComp-HCI: |
| 由来 |
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キーワード | ELECTRON TRANSPORT / Rieske-type aromatic dioxygenase |
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