Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing
Transglutaminases are a family of enzymes that catalyse the cross‐linking of proteins by forming covalent bonds between lysine and glutamine residues in various polypeptides. Cross‐linking reactions are involved in blood clots, skin formation, embryogenesis and apoptosis. Clinically, these enzymes a...
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FEBS Press and John Wiley & Sons Ltd
2020
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Online Access: | https://hdl.handle.net/11250/2731324 https://doi.org/10.1002/2211-5463.12826 |
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ftunivstavanger:oai:uis.brage.unit.no:11250/2731324 2023-06-11T04:10:10+02:00 Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing Álvarez, Rebeca García Karki, Pralav Langleite, Ida Elise Bakksjø, Ragna-Johanne Eichacker, Lutz Andreas Furnes, Clemens 2020-09-23T13:53:26Z application/pdf https://hdl.handle.net/11250/2731324 https://doi.org/10.1002/2211-5463.12826 eng eng FEBS Press and John Wiley & Sons Ltd Universitetet i Stavanger: 8790 Garcia Alvarez, R., Karki, P., Langli, E. et al. (2020) Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing. FEBS Open Bio, 10(4), 495-506. urn:issn:2211-5463 https://hdl.handle.net/11250/2731324 https://doi.org/10.1002/2211-5463.12826 cristin:1832586 Navngivelse 4.0 Internasjonal http://creativecommons.org/licenses/by/4.0/deed.no © 2020 The Authors. 495-506 10 FEBS Open Bio 4 VDP::Matematikk og Naturvitenskap: 400::Basale biofag: 470 Peer reviewed Journal article 2020 ftunivstavanger https://doi.org/10.1002/2211-5463.12826 2023-05-29T16:03:35Z Transglutaminases are a family of enzymes that catalyse the cross‐linking of proteins by forming covalent bonds between lysine and glutamine residues in various polypeptides. Cross‐linking reactions are involved in blood clots, skin formation, embryogenesis and apoptosis. Clinically, these enzymes appear to be implicated in neurodegenerative diseases, tumours and coeliac diseases. Transglutaminases have great potential for use in the food industry because of their ability to cross‐link proteins that are not normally linked. Here, a gene coding for transglutaminase from Atlantic cod was cloned into a bacterial expression vector and used to transform protein expression in a strain of Escherichia coli. The successful expression of recombinant transglutaminase protein from Atlantic cod (AcTG‐1) as a soluble protein upon induction at low temperature was confirmed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis, immunoblotting and mass spectrometry analysis. Biochemical characterisation demonstrated that the transglutaminase was active between 0 and 65 °C, but was completely inactivated after 20‐min incubation at 70 °C. Interestingly, the enzyme displayed cold‐adapted features, such as temperature instability combined with high catalytic efficiency at low temperatures (8–16 °C). In addition, the enzyme had optimal activity at 50 °C, a new feature for a cold‐adapted enzyme. AcTG‐1 was active in the pH range from 6 to 9, with an optimum at pH 8, and required 5 mm calcium for maximum activity. Potential calcium‐binding sites in the enzyme were predictable, making the enzyme an appropriate model for studying structure–function relationships in the calcium‐dependent transglutaminase family. In vitro gel analysis revealed that transglutaminase cross‐linked casein, collagen and gelatin. The binding of fish fillets in the presence of recombinant AcTG‐1 provided further macroscopic proof for the potential application of AcTG‐1 as a biological cross‐linker in the food industry. Once binding occurred, fish fillets ... Article in Journal/Newspaper atlantic cod University of Stavanger: UiS Brage FEBS Open Bio 10 4 495 506 |
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Open Polar |
collection |
University of Stavanger: UiS Brage |
op_collection_id |
ftunivstavanger |
language |
English |
topic |
VDP::Matematikk og Naturvitenskap: 400::Basale biofag: 470 |
spellingShingle |
VDP::Matematikk og Naturvitenskap: 400::Basale biofag: 470 Álvarez, Rebeca García Karki, Pralav Langleite, Ida Elise Bakksjø, Ragna-Johanne Eichacker, Lutz Andreas Furnes, Clemens Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
topic_facet |
VDP::Matematikk og Naturvitenskap: 400::Basale biofag: 470 |
description |
Transglutaminases are a family of enzymes that catalyse the cross‐linking of proteins by forming covalent bonds between lysine and glutamine residues in various polypeptides. Cross‐linking reactions are involved in blood clots, skin formation, embryogenesis and apoptosis. Clinically, these enzymes appear to be implicated in neurodegenerative diseases, tumours and coeliac diseases. Transglutaminases have great potential for use in the food industry because of their ability to cross‐link proteins that are not normally linked. Here, a gene coding for transglutaminase from Atlantic cod was cloned into a bacterial expression vector and used to transform protein expression in a strain of Escherichia coli. The successful expression of recombinant transglutaminase protein from Atlantic cod (AcTG‐1) as a soluble protein upon induction at low temperature was confirmed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis, immunoblotting and mass spectrometry analysis. Biochemical characterisation demonstrated that the transglutaminase was active between 0 and 65 °C, but was completely inactivated after 20‐min incubation at 70 °C. Interestingly, the enzyme displayed cold‐adapted features, such as temperature instability combined with high catalytic efficiency at low temperatures (8–16 °C). In addition, the enzyme had optimal activity at 50 °C, a new feature for a cold‐adapted enzyme. AcTG‐1 was active in the pH range from 6 to 9, with an optimum at pH 8, and required 5 mm calcium for maximum activity. Potential calcium‐binding sites in the enzyme were predictable, making the enzyme an appropriate model for studying structure–function relationships in the calcium‐dependent transglutaminase family. In vitro gel analysis revealed that transglutaminase cross‐linked casein, collagen and gelatin. The binding of fish fillets in the presence of recombinant AcTG‐1 provided further macroscopic proof for the potential application of AcTG‐1 as a biological cross‐linker in the food industry. Once binding occurred, fish fillets ... |
format |
Article in Journal/Newspaper |
author |
Álvarez, Rebeca García Karki, Pralav Langleite, Ida Elise Bakksjø, Ragna-Johanne Eichacker, Lutz Andreas Furnes, Clemens |
author_facet |
Álvarez, Rebeca García Karki, Pralav Langleite, Ida Elise Bakksjø, Ragna-Johanne Eichacker, Lutz Andreas Furnes, Clemens |
author_sort |
Álvarez, Rebeca García |
title |
Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
title_short |
Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
title_full |
Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
title_fullStr |
Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
title_full_unstemmed |
Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
title_sort |
characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing |
publisher |
FEBS Press and John Wiley & Sons Ltd |
publishDate |
2020 |
url |
https://hdl.handle.net/11250/2731324 https://doi.org/10.1002/2211-5463.12826 |
genre |
atlantic cod |
genre_facet |
atlantic cod |
op_source |
495-506 10 FEBS Open Bio 4 |
op_relation |
Universitetet i Stavanger: 8790 Garcia Alvarez, R., Karki, P., Langli, E. et al. (2020) Characterisation of a novel cold-adapted calcium- activated transglutaminase: implications for medicine and food processing. FEBS Open Bio, 10(4), 495-506. urn:issn:2211-5463 https://hdl.handle.net/11250/2731324 https://doi.org/10.1002/2211-5463.12826 cristin:1832586 |
op_rights |
Navngivelse 4.0 Internasjonal http://creativecommons.org/licenses/by/4.0/deed.no © 2020 The Authors. |
op_doi |
https://doi.org/10.1002/2211-5463.12826 |
container_title |
FEBS Open Bio |
container_volume |
10 |
container_issue |
4 |
container_start_page |
495 |
op_container_end_page |
506 |
_version_ |
1768384420528521216 |