Characterization of cold-adapted Atlantic cod (Gadus morhua) trypsin I — Kinetic parameters, autolysis and thermal stability

Stefansson, Bjarki; Helgadóttir, Linda; Ólafsdóttir, Sigríður; Guðmundsdóttir, Ágústa; Bjarnason, Jón B.

doi:10.1016/j.cbpb.2009.11.004

Cited by 34 publications

(48 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Multiple numbers of these amino acids are frequently found within proteins that can explain the deactivation of a wide spectrum of viruses by ColdZyme. Trypsin from Atlantic cod (Gadus morhua) has been shown to have higher catalytic efficiency than comparable enzymes [11][12][13][14][15]. Furthermore, native proteins, such as those found on the surface of viruses, appear to be more readily hydrolyzed by cod trypsin [15].…”

Section: Discussionmentioning

confidence: 99%

A medical device forming a protective barrier that deactivates four major common cold viruses

Stefansson¹,

Guðmundsdóttir²,

Clarsund³

2017

Virol Res Rev

Self Cite

View full text Add to dashboard Cite

The medical device ColdZyme is a mouth spray that forms a barrier in the throat against common cold viruses. The barrier solution of the device is composed of glycerol and Atlantic cod trypsin. The aim of this study was to evaluate the virus deactivating ability of ColdZyme against four major common cold viruses. A virucidal efficacy suspension test was conducted using ColdZyme against each of the challenge viruses in suspension. ColdZyme deactivated rhinovirus type 1A by 91.7% (1.08 log 10 ), rhinovirus type 42 by 92.8% (1.14 log 10 ), human influenza A virus H3N2 by 96.9% (1.51 log 10 ), respiratory syncytial virus (RSV) by 99.9% (2.94 log 10 ) and adenovirus type 2 by 64.5% (0.45 log 10 ). Based on the results, ColdZyme showed an effective broad-spectrum impact against common cold viruses. Thus, ColdZyme might represent a device with clinical benefits in prevention and treatment of respiratory viral infections by deactivating viruses within the respiratory tract.

show abstract

Section: Discussionmentioning

confidence: 99%

A medical device forming a protective barrier that deactivates four major common cold viruses

Stefansson¹,

Guðmundsdóttir²,

Clarsund³

2017

Virol Res Rev

Self Cite

View full text Add to dashboard Cite

show abstract

“…Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was done according to Laemmli as previously described [3]. Silver staining was performed as previously described [17].…”

Section: Electrophoresismentioning

confidence: 99%

“…Two Atlantic cod trypsins termed I and X differ by eight amino acid residues as deduced from their cDNA sequences [5,6]. Of the different cod trypsin isoenzymes trypsin I is most abundant and is the only thoroughly characterized cod trypsin whereas trypsin X has not yet been well characterized [3,4]. For unknown reasons, cod trypsin I has proven to be difficult to produce recombinantly in bacteria [7] and yeast (unpublished results).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Elucidation of different cold-adapted Atlantic cod ( Gadus morhua ) trypsin X isoenzymes

Stefansson¹,

Sandholt

Guðmundsdóttir

2017

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

Self Cite

View full text Add to dashboard Cite

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 (k cat /K m ) 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.

show abstract

“…Especially, fish trypsin displays substantially higher kcat/Km value at low temperatures than their mammalian counterparts. For example, the catalytic efficiency of cod trypsin was found to be higher than that of bovine trypsin [14,15], and that of salmon anionic trypsin was reported to be as much as 40-fold higher [16]. In addition, fish trypsin is more sensitive to inactivation by heat, low pH and autolysis than that of mesophilic analogues [14,17].…”

Section: Introductionmentioning

confidence: 99%

Structural properties of trypsin from cold-adapted fish, arabesque greenling (Pleurogrammus azonus)

Kanno

Kishimura

Ando

et al. 2010

Eur Food Res Technol

View full text Add to dashboard Cite

A cDNA clone encoding trypsin (AG-T) was isolated from the pyloric ceca of cold-adapted fish, arabesque greenling (Pleurogrammus azonus). The cDNA was composed of 892 bp with an open reading frame of 729 bp at nucleotide positions 25-753. Similar to all the known trypsin, the AG-T seemed to be synthesized as preproenzyme that contains a hydrophobic signal peptide, an activation pentapeptide and a mature trypsin of 222 amino acid residues. The AG-T also completely conserved the major structural features common to trypsin such as the catalytic triad (His57, Asp102 and Ser195), the obligatory Asp189 and twelve Cys residues. On the other hand, the AG-T possessed the deletion of Tyr151 and substitution of Pro152 for Gly in the autolysis loop when aligned with the sequence of tropical-zone fish and bovine trypsins. In addition, Val75 concerned in a combination with calcium ion was exchanged for Ala in the AG-T and the content of positively charged amino acid residues at the calcium-binding site of the AG-T was three times higher than those of tropical-zone fish trypsins. Moreover, the ratio between charged and hydrophobic amino acid residues in the N-terminal region of the AG-T was also higher than those of temperate-zone fish and tropical-zone fish trypsins. Such structural properties of the AG-T would contribute to its low thermostability.

show abstract

Characterization of cold-adapted Atlantic cod (Gadus morhua) trypsin I — Kinetic parameters, autolysis and thermal stability

Cited by 34 publications

References 42 publications

A medical device forming a protective barrier that deactivates four major common cold viruses

A medical device forming a protective barrier that deactivates four major common cold viruses

Elucidation of different cold-adapted Atlantic cod ( Gadus morhua ) trypsin X isoenzymes

Structural properties of trypsin from cold-adapted fish, arabesque greenling (Pleurogrammus azonus)

Contact Info

Product

Resources

About