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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

Authors

Matteo Planchestainer matteo.planchestainer@nottingham.ac.uk

David Roura Padrosa pcxdr1@exmail.nottingham.ac.uk

Martina Letizia Contente



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

Citation

Planchestainer, M., Padrosa, D. R., Contente, M. L., & Paradisi, F. (2018). Genetically fused T4L acts as a shield in covalent enzyme immobilisation enhancing the rescued activity. Catalysts, 8(1), https://doi.org/10.3390/catal8010040

Journal Article Type Article
Acceptance Date Jan 16, 2018
Publication Date Jan 20, 2018
Deposit Date Jan 31, 2018
Publicly Available Date Jan 31, 2018
Journal Catalysts
Electronic ISSN 2073-4344
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 8
Issue 1
DOI https://doi.org/10.3390/catal8010040
Keywords aminotransferase; esterase; alcohol dehydrogenases; biocatalysis; enzyme immobilisation
Public URL http://eprints.nottingham.ac.uk/id/eprint/49456
Publisher URL http://www.mdpi.com/2073-4344/8/1/40
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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