Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression
Following a screening of Antarctic glacier forefield-bacteria for novel cold-active enzymes, a psychrophilic strain Psychrobacter sp. 94-6PB was selected for further characterization of enzymatic activities. The strain produced lipases and proteases in the temperature range of 4–18°C. The coding seq...
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ftpubmed:oai:pubmedcentral.nih.gov:7247812 2023-05-15T14:00:03+02:00 Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression Perfumo, Amedea Freiherr von Sass, Georg Johannes Nordmann, Eva-Lena Budisa, Nediljko Wagner, Dirk 2020-05-13 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7247812/ https://doi.org/10.3389/fmicb.2020.00881 en eng Frontiers Media S.A. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7247812/ http://dx.doi.org/10.3389/fmicb.2020.00881 Copyright © 2020 Perfumo, Freiherr von Sass, Nordmann, Budisa and Wagner. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. CC-BY Front Microbiol Microbiology Text 2020 ftpubmed https://doi.org/10.3389/fmicb.2020.00881 2020-06-14T00:20:23Z Following a screening of Antarctic glacier forefield-bacteria for novel cold-active enzymes, a psychrophilic strain Psychrobacter sp. 94-6PB was selected for further characterization of enzymatic activities. The strain produced lipases and proteases in the temperature range of 4–18°C. The coding sequence of an extracellular serine-protease was then identified via comparative analysis across Psychrobacter sp. genomes, PCR-amplified in our strain 94-6PB and expressed in the heterologous host E. coli. The purified enzyme (80 kDa) resulted to be a cold-active alkaline protease, performing best at temperatures of 20–30°C and pH 7-9. It was stable in presence of common inhibitors [β-mercaptoethanol (β-ME), dithiothreitol (DTT), urea, phenylmethylsulfonyl fluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA)] and compatible with detergents and surfactants (Tween 20, Tween 80, hydrogen peroxide and Triton X-100). Because of these properties, the P94-6PB protease may be suitable for use in a new generation of laundry products for cold washing. Furthermore, we assessed the microdiversity of this enzyme in Psychrobacter organisms from different cold habitats and found several gene clusters that correlated with specific ecological niches. We then discussed the role of habitat specialization in shaping the biodiversity of proteins and enzymes and anticipate far-reaching implications for the search of novel variants of biotechnological products. Text Antarc* Antarctic PubMed Central (PMC) Antarctic Triton ENVELOPE(-55.615,-55.615,49.517,49.517) Frontiers in Microbiology 11 |
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PubMed Central (PMC) |
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ftpubmed |
language |
English |
topic |
Microbiology |
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Microbiology Perfumo, Amedea Freiherr von Sass, Georg Johannes Nordmann, Eva-Lena Budisa, Nediljko Wagner, Dirk Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
topic_facet |
Microbiology |
description |
Following a screening of Antarctic glacier forefield-bacteria for novel cold-active enzymes, a psychrophilic strain Psychrobacter sp. 94-6PB was selected for further characterization of enzymatic activities. The strain produced lipases and proteases in the temperature range of 4–18°C. The coding sequence of an extracellular serine-protease was then identified via comparative analysis across Psychrobacter sp. genomes, PCR-amplified in our strain 94-6PB and expressed in the heterologous host E. coli. The purified enzyme (80 kDa) resulted to be a cold-active alkaline protease, performing best at temperatures of 20–30°C and pH 7-9. It was stable in presence of common inhibitors [β-mercaptoethanol (β-ME), dithiothreitol (DTT), urea, phenylmethylsulfonyl fluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA)] and compatible with detergents and surfactants (Tween 20, Tween 80, hydrogen peroxide and Triton X-100). Because of these properties, the P94-6PB protease may be suitable for use in a new generation of laundry products for cold washing. Furthermore, we assessed the microdiversity of this enzyme in Psychrobacter organisms from different cold habitats and found several gene clusters that correlated with specific ecological niches. We then discussed the role of habitat specialization in shaping the biodiversity of proteins and enzymes and anticipate far-reaching implications for the search of novel variants of biotechnological products. |
format |
Text |
author |
Perfumo, Amedea Freiherr von Sass, Georg Johannes Nordmann, Eva-Lena Budisa, Nediljko Wagner, Dirk |
author_facet |
Perfumo, Amedea Freiherr von Sass, Georg Johannes Nordmann, Eva-Lena Budisa, Nediljko Wagner, Dirk |
author_sort |
Perfumo, Amedea |
title |
Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
title_short |
Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
title_full |
Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
title_fullStr |
Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
title_full_unstemmed |
Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression |
title_sort |
discovery and characterization of a new cold-active protease from an extremophilic bacterium via comparative genome analysis and in vitro expression |
publisher |
Frontiers Media S.A. |
publishDate |
2020 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7247812/ https://doi.org/10.3389/fmicb.2020.00881 |
long_lat |
ENVELOPE(-55.615,-55.615,49.517,49.517) |
geographic |
Antarctic Triton |
geographic_facet |
Antarctic Triton |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Front Microbiol |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7247812/ http://dx.doi.org/10.3389/fmicb.2020.00881 |
op_rights |
Copyright © 2020 Perfumo, Freiherr von Sass, Nordmann, Budisa and Wagner. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fmicb.2020.00881 |
container_title |
Frontiers in Microbiology |
container_volume |
11 |
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