Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes
Trypsins from Atlantic cod (Gadus morhua), consisting of several isoenzymes, are highly active cold-adapted serine proteases. These trypsins are isolated for biomedical use in an eco-friendly manner from underutilized seafood by-products. Our group has explored the biochemical properties of trypsins...
Published in: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics |
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Online Access: | https://hdl.handle.net/20.500.11815/153 https://doi.org/10.1016/j.bbapap.2016.10.005 |
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ftopinvisindi:oai:opinvisindi.is:20.500.11815/153 2024-09-15T17:55:22+00:00 Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes Stefánsson, Bjarki Sandholt, Gunnar Birgir Guðmundsdóttir, Ágústa Matvæla- og næringarfræðideild (HÍ) Faculty of Food Science and Nutrition (UI) Heilbrigðisvísindasvið (HÍ) School of Health Sciences (UI) Háskóli Íslands University of Iceland 2017-01 11-19 https://hdl.handle.net/20.500.11815/153 https://doi.org/10.1016/j.bbapap.2016.10.005 en eng Elsevier BV Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics;1865(1) Stefansson, B., Sandholt, G. B., & Gudmundsdottir, Á. (2017). Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 1865(1), 11–19. doi:10.1016/j.bbapap.2016.10.005 1570-9639 https://hdl.handle.net/20.500.11815/153 Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics doi:10.1016/j.bbapap.2016.10.005 info:eu-repo/semantics/openAccess Biophysics Analytical Chemistry Biochemistry Molecular Biology Lífefnafræði Efnagreining Lífeðlisfræði Sameindalíffræði Biomedicine Lífvísindi Atlantic cod Atlantshafslax info:eu-repo/semantics/article 2017 ftopinvisindi https://doi.org/20.500.11815/15310.1016/j.bbapap.2016.10.005 2024-07-09T03:01:56Z Trypsins from Atlantic cod (Gadus morhua), consisting of several isoenzymes, are highly active cold-adapted serine proteases. These trypsins are isolated for biomedical use in an eco-friendly manner from underutilized seafood by-products. Our group has explored the biochemical properties of trypsins and their high potential in biomedicine. For broader utilization of cod trypsins, further characterization of biochemical properties of the individual cod trypsin isoenzymes is of importance. For that purpose, a benzamidine purified trypsin isolate from Atlantic cod was analyzed. Anion exchange chromatography revealed eight peaks containing proteins around 24 kDa with tryptic activity. Based on mass spectrometric analysis, one isoenzyme gave the best match to cod trypsin I and six isoenzymes gave the best match to cod trypsin X. Amino terminal sequencing of two of these six trypsin isoenzymes showed identity to cod trypsin X. Three sequence variants of trypsin X were identified by cDNA analysis demonstrating that various forms of this enzyme exist. One trypsin X isoenzyme was selected for further characterization based on abundance and stability. Stepwise increase in catalytic efficiency (kcat/Km) of this trypsin X isoenzyme was obtained with substrates containing one to three amino acid residues. The study demonstrates that the catalytic efficiency of this trypsin X isoenzyme is comparable to that of cod trypsin I, the most abundant and highly active isoenzyme in the benzamidine cod trypsin isolate. Differences in pH stability and sensitivity to inhibitors of the trypsin X isoenzyme compared to cod trypsin I were detected that may be important for practical use. Research funded by AVS R&D Fund of Ministry of Fisheries and Agriculture in Iceland (R069-08, R11 028-11, R14 044-14) | Science and Technology Development Fund (120852-0611, 131804-0611) Ritrýnt tímarit Peer Reviewed Pre-Print Article in Journal/Newspaper atlantic cod Gadus morhua Iceland Opin vísindi (Iceland) Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 1865 1 11 19 |
institution |
Open Polar |
collection |
Opin vísindi (Iceland) |
op_collection_id |
ftopinvisindi |
language |
English |
topic |
Biophysics Analytical Chemistry Biochemistry Molecular Biology Lífefnafræði Efnagreining Lífeðlisfræði Sameindalíffræði Biomedicine Lífvísindi Atlantic cod Atlantshafslax |
spellingShingle |
Biophysics Analytical Chemistry Biochemistry Molecular Biology Lífefnafræði Efnagreining Lífeðlisfræði Sameindalíffræði Biomedicine Lífvísindi Atlantic cod Atlantshafslax Stefánsson, Bjarki Sandholt, Gunnar Birgir Guðmundsdóttir, Ágústa Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
topic_facet |
Biophysics Analytical Chemistry Biochemistry Molecular Biology Lífefnafræði Efnagreining Lífeðlisfræði Sameindalíffræði Biomedicine Lífvísindi Atlantic cod Atlantshafslax |
description |
Trypsins from Atlantic cod (Gadus morhua), consisting of several isoenzymes, are highly active cold-adapted serine proteases. These trypsins are isolated for biomedical use in an eco-friendly manner from underutilized seafood by-products. Our group has explored the biochemical properties of trypsins and their high potential in biomedicine. For broader utilization of cod trypsins, further characterization of biochemical properties of the individual cod trypsin isoenzymes is of importance. For that purpose, a benzamidine purified trypsin isolate from Atlantic cod was analyzed. Anion exchange chromatography revealed eight peaks containing proteins around 24 kDa with tryptic activity. Based on mass spectrometric analysis, one isoenzyme gave the best match to cod trypsin I and six isoenzymes gave the best match to cod trypsin X. Amino terminal sequencing of two of these six trypsin isoenzymes showed identity to cod trypsin X. Three sequence variants of trypsin X were identified by cDNA analysis demonstrating that various forms of this enzyme exist. One trypsin X isoenzyme was selected for further characterization based on abundance and stability. Stepwise increase in catalytic efficiency (kcat/Km) of this trypsin X isoenzyme was obtained with substrates containing one to three amino acid residues. The study demonstrates that the catalytic efficiency of this trypsin X isoenzyme is comparable to that of cod trypsin I, the most abundant and highly active isoenzyme in the benzamidine cod trypsin isolate. Differences in pH stability and sensitivity to inhibitors of the trypsin X isoenzyme compared to cod trypsin I were detected that may be important for practical use. Research funded by AVS R&D Fund of Ministry of Fisheries and Agriculture in Iceland (R069-08, R11 028-11, R14 044-14) | Science and Technology Development Fund (120852-0611, 131804-0611) Ritrýnt tímarit Peer Reviewed Pre-Print |
author2 |
Matvæla- og næringarfræðideild (HÍ) Faculty of Food Science and Nutrition (UI) Heilbrigðisvísindasvið (HÍ) School of Health Sciences (UI) Háskóli Íslands University of Iceland |
format |
Article in Journal/Newspaper |
author |
Stefánsson, Bjarki Sandholt, Gunnar Birgir Guðmundsdóttir, Ágústa |
author_facet |
Stefánsson, Bjarki Sandholt, Gunnar Birgir Guðmundsdóttir, Ágústa |
author_sort |
Stefánsson, Bjarki |
title |
Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
title_short |
Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
title_full |
Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
title_fullStr |
Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
title_full_unstemmed |
Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes |
title_sort |
elucidation of different cold-adapted atlantic cod (gadus morhua) trypsin x isoenzymes |
publisher |
Elsevier BV |
publishDate |
2017 |
url |
https://hdl.handle.net/20.500.11815/153 https://doi.org/10.1016/j.bbapap.2016.10.005 |
genre |
atlantic cod Gadus morhua Iceland |
genre_facet |
atlantic cod Gadus morhua Iceland |
op_relation |
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics;1865(1) Stefansson, B., Sandholt, G. B., & Gudmundsdottir, Á. (2017). Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 1865(1), 11–19. doi:10.1016/j.bbapap.2016.10.005 1570-9639 https://hdl.handle.net/20.500.11815/153 Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics doi:10.1016/j.bbapap.2016.10.005 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/20.500.11815/15310.1016/j.bbapap.2016.10.005 |
container_title |
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics |
container_volume |
1865 |
container_issue |
1 |
container_start_page |
11 |
op_container_end_page |
19 |
_version_ |
1810431663457435648 |