The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis)
Inge W. Nilsen et al. Background We have previously isolated a thermolabile nuclease specific for double-stranded DNA from industrial processing water of Northern shrimps (Pandalus borealis) and developed an application of the enzyme in removal of contaminating DNA in PCR-related technologies. Metho...
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ftcsic:oai:digital.csic.es:10261/44040 2024-02-11T10:05:25+01:00 The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) Nilsen, Inge W. Lanes, Olav Lalueza-Fox, Carles 2010-04-22 http://hdl.handle.net/10261/44040 https://doi.org/10.1371/journal.pone.0010295 en eng Public Library of Science Publisher’s version http://dx.doi.org/10.1371/journal.pone.0010295 PLoS ONE 5(4): e10295 (2010) 1932-6203 http://hdl.handle.net/10261/44040 doi:10.1371/journal.pone.0010295 20421970 open artículo http://purl.org/coar/resource_type/c_6501 2010 ftcsic https://doi.org/10.1371/journal.pone.0010295 2024-01-16T09:35:20Z Inge W. Nilsen et al. Background We have previously isolated a thermolabile nuclease specific for double-stranded DNA from industrial processing water of Northern shrimps (Pandalus borealis) and developed an application of the enzyme in removal of contaminating DNA in PCR-related technologies. Methodology/Principal Findings A 43 kDa nuclease with a high specific activity of hydrolysing linear as well as circular forms of DNA was purified from hepatopancreas of Northern shrimp (Pandalus borealis). The enzyme displayed a substrate preference that was shifted from exclusively double-stranded DNA in the presence of magnesium to also encompass significant activity against single-stranded DNA when calcium was added. No activity against RNA was detected. Although originating from a cold-environment animal, the shrimp DNase has only minor low-temperature activity. Still, the enzyme was irreversibly inactivated by moderate heating with a half-life of 1 min at 65°C. The purified protein was partly sequenced and derived oligonucleotides were used to prime amplification of the encoding cDNA. This cDNA sequence revealed an open reading frame encoding a 404 amino acid protein containing a signal peptide. By sequence similarity the enzyme is predicted to belong to a family of DNA/RNA non-specific nucleases even though this shrimp DNase lacks RNase activity and is highly double-strand specific in some respects. These features are in agreement with those previously established for endonucleases classified as similar to the Kamchatka crab duplex-specific nuclease (Par_DSN). Sequence comparisons and phylogenetic analyses confirmed that the Northern shrimp nuclease resembles the Par_DSN-like nucleases and displays a more distant relationship to the Serratia family of nucleases. Conclusions/Significance The shrimp nuclease contains enzyme activity that may be controlled by temperature or buffer compositions. The double-stranded DNA specificity, as well as the thermolabile feature, strengthens its potential for in vitro ... Article in Journal/Newspaper Kamchatka Kamchatka crab northern shrimp Pandalus borealis Digital.CSIC (Spanish National Research Council) PLoS ONE 5 4 e10295 |
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Digital.CSIC (Spanish National Research Council) |
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English |
description |
Inge W. Nilsen et al. Background We have previously isolated a thermolabile nuclease specific for double-stranded DNA from industrial processing water of Northern shrimps (Pandalus borealis) and developed an application of the enzyme in removal of contaminating DNA in PCR-related technologies. Methodology/Principal Findings A 43 kDa nuclease with a high specific activity of hydrolysing linear as well as circular forms of DNA was purified from hepatopancreas of Northern shrimp (Pandalus borealis). The enzyme displayed a substrate preference that was shifted from exclusively double-stranded DNA in the presence of magnesium to also encompass significant activity against single-stranded DNA when calcium was added. No activity against RNA was detected. Although originating from a cold-environment animal, the shrimp DNase has only minor low-temperature activity. Still, the enzyme was irreversibly inactivated by moderate heating with a half-life of 1 min at 65°C. The purified protein was partly sequenced and derived oligonucleotides were used to prime amplification of the encoding cDNA. This cDNA sequence revealed an open reading frame encoding a 404 amino acid protein containing a signal peptide. By sequence similarity the enzyme is predicted to belong to a family of DNA/RNA non-specific nucleases even though this shrimp DNase lacks RNase activity and is highly double-strand specific in some respects. These features are in agreement with those previously established for endonucleases classified as similar to the Kamchatka crab duplex-specific nuclease (Par_DSN). Sequence comparisons and phylogenetic analyses confirmed that the Northern shrimp nuclease resembles the Par_DSN-like nucleases and displays a more distant relationship to the Serratia family of nucleases. Conclusions/Significance The shrimp nuclease contains enzyme activity that may be controlled by temperature or buffer compositions. The double-stranded DNA specificity, as well as the thermolabile feature, strengthens its potential for in vitro ... |
author2 |
Lalueza-Fox, Carles |
format |
Article in Journal/Newspaper |
author |
Nilsen, Inge W. Lanes, Olav |
spellingShingle |
Nilsen, Inge W. Lanes, Olav The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
author_facet |
Nilsen, Inge W. Lanes, Olav |
author_sort |
Nilsen, Inge W. |
title |
The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
title_short |
The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
title_full |
The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
title_fullStr |
The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
title_full_unstemmed |
The Enzyme and the cDNA Sequence of a Thermolabile and Double-Strand Specific DNase from Northern Shrimps (Pandalus borealis) |
title_sort |
enzyme and the cdna sequence of a thermolabile and double-strand specific dnase from northern shrimps (pandalus borealis) |
publisher |
Public Library of Science |
publishDate |
2010 |
url |
http://hdl.handle.net/10261/44040 https://doi.org/10.1371/journal.pone.0010295 |
genre |
Kamchatka Kamchatka crab northern shrimp Pandalus borealis |
genre_facet |
Kamchatka Kamchatka crab northern shrimp Pandalus borealis |
op_relation |
Publisher’s version http://dx.doi.org/10.1371/journal.pone.0010295 PLoS ONE 5(4): e10295 (2010) 1932-6203 http://hdl.handle.net/10261/44040 doi:10.1371/journal.pone.0010295 20421970 |
op_rights |
open |
op_doi |
https://doi.org/10.1371/journal.pone.0010295 |
container_title |
PLoS ONE |
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5 |
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4 |
container_start_page |
e10295 |
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1790602443347722240 |