The enzyme and the cDNA sequence of a thermolabile and double-strand specific DNase from Northern shrimps (Pandalus borealis).
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 FIND...
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ftdoajarticles:oai:doaj.org/article:525c647dc26041eea40a8b3fb69ceaa5 2023-05-15T16:59:26+02:00 The enzyme and the cDNA sequence of a thermolabile and double-strand specific DNase from Northern shrimps (Pandalus borealis). Inge W Nilsen Kersti Øverbø Linda Jensen Havdalen Morten Elde Dag Rune Gjellesvik Olav Lanes 2010-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0010295 https://doaj.org/article/525c647dc26041eea40a8b3fb69ceaa5 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC2858651?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0010295 https://doaj.org/article/525c647dc26041eea40a8b3fb69ceaa5 PLoS ONE, Vol 5, Iss 4, p e10295 (2010) Medicine R Science Q article 2010 ftdoajarticles https://doi.org/10.1371/journal.pone.0010295 2022-12-31T07:03:44Z 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 degrees 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 applications. Article in Journal/Newspaper Kamchatka Kamchatka crab northern shrimp Pandalus borealis Directory of Open Access Journals: DOAJ Articles PLoS ONE 5 4 e10295 |
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Medicine R Science Q Inge W Nilsen Kersti Øverbø Linda Jensen Havdalen Morten Elde Dag Rune Gjellesvik Olav Lanes The enzyme and the cDNA sequence of a thermolabile and double-strand specific DNase from Northern shrimps (Pandalus borealis). |
topic_facet |
Medicine R Science Q |
description |
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 degrees 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 applications. |
format |
Article in Journal/Newspaper |
author |
Inge W Nilsen Kersti Øverbø Linda Jensen Havdalen Morten Elde Dag Rune Gjellesvik Olav Lanes |
author_facet |
Inge W Nilsen Kersti Øverbø Linda Jensen Havdalen Morten Elde Dag Rune Gjellesvik Olav Lanes |
author_sort |
Inge W Nilsen |
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 (PLoS) |
publishDate |
2010 |
url |
https://doi.org/10.1371/journal.pone.0010295 https://doaj.org/article/525c647dc26041eea40a8b3fb69ceaa5 |
genre |
Kamchatka Kamchatka crab northern shrimp Pandalus borealis |
genre_facet |
Kamchatka Kamchatka crab northern shrimp Pandalus borealis |
op_source |
PLoS ONE, Vol 5, Iss 4, p e10295 (2010) |
op_relation |
http://europepmc.org/articles/PMC2858651?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0010295 https://doaj.org/article/525c647dc26041eea40a8b3fb69ceaa5 |
op_doi |
https://doi.org/10.1371/journal.pone.0010295 |
container_title |
PLoS ONE |
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5 |
container_issue |
4 |
container_start_page |
e10295 |
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1766051688500690944 |