Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf
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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ftfrontimediafig:oai:figshare.com:article/12292217 2023-05-15T13:32:08+02:00 Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf Amedea Perfumo Georg Johannes Freiherr von Sass Eva-Lena Nordmann Nediljko Budisa Dirk Wagner 2020-05-13T04:14:20Z https://doi.org/10.3389/fmicb.2020.00881.s001 https://figshare.com/articles/Data_Sheet_1_Discovery_and_Characterization_of_a_New_Cold-Active_Protease_From_an_Extremophilic_Bacterium_via_Comparative_Genome_Analysis_and_in_vitro_Expression_pdf/12292217 unknown doi:10.3389/fmicb.2020.00881.s001 https://figshare.com/articles/Data_Sheet_1_Discovery_and_Characterization_of_a_New_Cold-Active_Protease_From_an_Extremophilic_Bacterium_via_Comparative_Genome_Analysis_and_in_vitro_Expression_pdf/12292217 CC BY 4.0 CC-BY Microbiology Microbial Genetics Microbial Ecology Mycology bioprospecting extremophilic bacteria cold-active enzymes genome mining heterologous protein expression microdiversity Dataset 2020 ftfrontimediafig https://doi.org/10.3389/fmicb.2020.00881.s001 2020-05-13T22:53:13Z 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. Dataset Antarc* Antarctic Frontiers: Figshare Antarctic Triton ENVELOPE(-55.615,-55.615,49.517,49.517) |
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Open Polar |
collection |
Frontiers: Figshare |
op_collection_id |
ftfrontimediafig |
language |
unknown |
topic |
Microbiology Microbial Genetics Microbial Ecology Mycology bioprospecting extremophilic bacteria cold-active enzymes genome mining heterologous protein expression microdiversity |
spellingShingle |
Microbiology Microbial Genetics Microbial Ecology Mycology bioprospecting extremophilic bacteria cold-active enzymes genome mining heterologous protein expression microdiversity Amedea Perfumo Georg Johannes Freiherr von Sass Eva-Lena Nordmann Nediljko Budisa Dirk Wagner Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
topic_facet |
Microbiology Microbial Genetics Microbial Ecology Mycology bioprospecting extremophilic bacteria cold-active enzymes genome mining heterologous protein expression microdiversity |
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 |
Dataset |
author |
Amedea Perfumo Georg Johannes Freiherr von Sass Eva-Lena Nordmann Nediljko Budisa Dirk Wagner |
author_facet |
Amedea Perfumo Georg Johannes Freiherr von Sass Eva-Lena Nordmann Nediljko Budisa Dirk Wagner |
author_sort |
Amedea Perfumo |
title |
Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
title_short |
Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
title_full |
Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
title_fullStr |
Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
title_full_unstemmed |
Data_Sheet_1_Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression.pdf |
title_sort |
data_sheet_1_discovery and characterization of a new cold-active protease from an extremophilic bacterium via comparative genome analysis and in vitro expression.pdf |
publishDate |
2020 |
url |
https://doi.org/10.3389/fmicb.2020.00881.s001 https://figshare.com/articles/Data_Sheet_1_Discovery_and_Characterization_of_a_New_Cold-Active_Protease_From_an_Extremophilic_Bacterium_via_Comparative_Genome_Analysis_and_in_vitro_Expression_pdf/12292217 |
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_relation |
doi:10.3389/fmicb.2020.00881.s001 https://figshare.com/articles/Data_Sheet_1_Discovery_and_Characterization_of_a_New_Cold-Active_Protease_From_an_Extremophilic_Bacterium_via_Comparative_Genome_Analysis_and_in_vitro_Expression_pdf/12292217 |
op_rights |
CC BY 4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fmicb.2020.00881.s001 |
_version_ |
1766024491525210112 |