Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity
Enzyme immobilisation is a common strategy to increase enzymes resistance and reusability in a variety of excellent ‘green’ applications. However, the interaction with the solid support often leads to diminished specific activity, especially when non-specific covalent binding to the carrier takes pl...
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| Format: | Article |
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MDPI
2018
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| Online Access: | https://eprints.nottingham.ac.uk/49456/ |
| _version_ | 1848798000206315520 |
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| author | Planchestainer, Matteo Padrosa, David Roura Contente, Martina Letizia Paradisi, Francesca |
| author_facet | Planchestainer, Matteo Padrosa, David Roura Contente, Martina Letizia Paradisi, Francesca |
| author_sort | Planchestainer, Matteo |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Enzyme immobilisation is a common strategy to increase enzymes resistance and reusability in a variety of excellent ‘green’ applications. However, the interaction with the solid support often leads to diminished specific activity, especially when non-specific covalent binding to the carrier takes place which affects the delicate architecture of the enzyme. Here we developed a broadly applicable strategy where the T4-lysozyme (T4L) is genetically fused at the N-terminus of different enzymes and used as inert protein spacer which directly attaches to the carrier preventing shape distortion of the catalyst. Halomonas elongata aminotransferase (HEWT), Bacillus subtilis engineered esterase (BS2m), and horse liver alcohol dehydrogenase (HLADH) were used as model enzymes to elucidate the benefits of the spacer. While HEWT and HLADH activity and expression were diminished by the fused T4L, both enzymes retained almost quantitative activity after immobilisation. In the case of BS2m, the protective effect of the T4L effectively was important and led to up to 10-fold improvement in the rescued activity |
| first_indexed | 2025-11-14T20:12:48Z |
| format | Article |
| id | nottingham-49456 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:12:48Z |
| publishDate | 2018 |
| publisher | MDPI |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-494562020-05-04T19:27:43Z https://eprints.nottingham.ac.uk/49456/ Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity Planchestainer, Matteo Padrosa, David Roura Contente, Martina Letizia Paradisi, Francesca Enzyme immobilisation is a common strategy to increase enzymes resistance and reusability in a variety of excellent ‘green’ applications. However, the interaction with the solid support often leads to diminished specific activity, especially when non-specific covalent binding to the carrier takes place which affects the delicate architecture of the enzyme. Here we developed a broadly applicable strategy where the T4-lysozyme (T4L) is genetically fused at the N-terminus of different enzymes and used as inert protein spacer which directly attaches to the carrier preventing shape distortion of the catalyst. Halomonas elongata aminotransferase (HEWT), Bacillus subtilis engineered esterase (BS2m), and horse liver alcohol dehydrogenase (HLADH) were used as model enzymes to elucidate the benefits of the spacer. While HEWT and HLADH activity and expression were diminished by the fused T4L, both enzymes retained almost quantitative activity after immobilisation. In the case of BS2m, the protective effect of the T4L effectively was important and led to up to 10-fold improvement in the rescued activity MDPI 2018-01-20 Article PeerReviewed Planchestainer, Matteo, Padrosa, David Roura, Contente, Martina Letizia and Paradisi, Francesca (2018) Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity. Catalysts, 8 (1). p. 40. ISSN 2073-4344 aminotransferase; esterase; alcohol dehydrogenases; biocatalysis; enzyme immobilisation http://www.mdpi.com/2073-4344/8/1/40 doi:10.3390/catal8010040 doi:10.3390/catal8010040 |
| spellingShingle | aminotransferase; esterase; alcohol dehydrogenases; biocatalysis; enzyme immobilisation Planchestainer, Matteo Padrosa, David Roura Contente, Martina Letizia Paradisi, Francesca Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title | Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title_full | Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title_fullStr | Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title_full_unstemmed | Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title_short | Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| title_sort | genetically fused t4l acts as a shield in covalent enzyme immobilisation enhancing the rescued activity |
| topic | aminotransferase; esterase; alcohol dehydrogenases; biocatalysis; enzyme immobilisation |
| url | https://eprints.nottingham.ac.uk/49456/ https://eprints.nottingham.ac.uk/49456/ https://eprints.nottingham.ac.uk/49456/ |