Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity

Planchestainer, Matteo and Padrosa, David Roura and 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

[img]
Preview
PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Available under Licence Creative Commons Attribution.
Download (1MB) | Preview

Abstract

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

Item Type: Article
Keywords: aminotransferase; esterase; alcohol dehydrogenases; biocatalysis; enzyme immobilisation
Schools/Departments: University of Nottingham, UK > Faculty of Science > School of Chemistry
Identification Number: https://doi.org/10.3390/catal8010040
Depositing User: Eprints, Support
Date Deposited: 31 Jan 2018 14:15
Last Modified: 02 Jul 2018 09:18
URI: http://eprints.nottingham.ac.uk/id/eprint/49456

Actions (Archive Staff Only)

Edit View Edit View