Academic Journal
Mutations Closer to the Active Site Improve the Promiscuous Aldolase Activity of 4-Oxalocrotonate Tautomerase More Effectively than Distant Mutations
| العنوان: | Mutations Closer to the Active Site Improve the Promiscuous Aldolase Activity of 4-Oxalocrotonate Tautomerase More Effectively than Distant Mutations |
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| المؤلفون: | Rahimi, Mehran, van der Meer, Jan-Ytzen, Geertsema, Edzard M, Poddar, Harshwardhan, Baas, Bert-Jan, Poelarends, Gerrit J |
| المصدر: | Rahimi , M , van der Meer , J-Y , Geertsema , E M , Poddar , H , Baas , B-J & Poelarends , G J 2016 , ' Mutations Closer to the Active Site Improve the Promiscuous Aldolase Activity of 4-Oxalocrotonate Tautomerase More Effectively than Distant Mutations ' , ChemBioChem , vol. 17 , no. 13 , pp. 1225-1228 . https://doi.org/10.1002/cbic.201600149 |
| سنة النشر: | 2016 |
| المجموعة: | University of Groningen research database |
| الوصف: | The enzyme 4-oxalocrotonate tautomerase (4-OT), which catalyzes enol-keto tautomerization as part of a degradative pathway for aromatic hydrocarbons, promiscuously catalyzes various carbon-carbon bond-forming reactions. These include the aldol condensation of acetaldehyde with benzaldehyde to yield cinnamaldehyde. Here, we demonstrate that 4-OT can be engineered into a more efficient aldolase for this condensation reaction, with a >5000-fold improvement in catalytic efficiency (kcat /Km ) and a >10(7) -fold change in reaction specificity, by exploring small libraries in which only "hotspots" are varied. The hotspots were identified by systematic mutagenesis (covering each residue), followed by a screen for single mutations that give a strong improvement in the desired aldolase activity. All beneficial mutations were near the active site of 4-OT, thus underpinning the notion that new catalytic activities of a promiscuous enzyme are more effectively enhanced by mutations close to the active site. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | https://research.rug.nl/en/publications/7761884a-961d-476e-bbdb-2485fbfa87c3 |
| DOI: | 10.1002/cbic.201600149 |
| الاتاحة: | https://hdl.handle.net/11370/7761884a-961d-476e-bbdb-2485fbfa87c3 https://research.rug.nl/en/publications/7761884a-961d-476e-bbdb-2485fbfa87c3 https://doi.org/10.1002/cbic.201600149 https://pure.rug.nl/ws/files/79156693/Mutations_Closer_to_the_Active_Site_Improve_the_Promiscuous_Aldolase_Activity.pdf |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.9324ADB7 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1002/cbic.201600149 |
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