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A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application
L-asparaginase (L-ASNase) is a vital enzyme with a broad range of applications in medicine and food industry. Drawbacks of current commercial L-ASNases stimulate the search for better-producing sources of the enzyme, and extremophiles are especially attractive in this view. In this study, a novel L-...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470970/ https://www.ncbi.nlm.nih.gov/pubmed/34576056 http://dx.doi.org/10.3390/ijms22189894 |
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author | Dumina, Maria Zhgun, Alexander Pokrovskaya, Marina Aleksandrova, Svetlana Zhdanov, Dmitry Sokolov, Nikolay El’darov, Michael |
author_facet | Dumina, Maria Zhgun, Alexander Pokrovskaya, Marina Aleksandrova, Svetlana Zhdanov, Dmitry Sokolov, Nikolay El’darov, Michael |
author_sort | Dumina, Maria |
collection | PubMed |
description | L-asparaginase (L-ASNase) is a vital enzyme with a broad range of applications in medicine and food industry. Drawbacks of current commercial L-ASNases stimulate the search for better-producing sources of the enzyme, and extremophiles are especially attractive in this view. In this study, a novel L-asparaginase originating from the hyperthermophilic archaeon Thermococcus sibiricus (TsA) was expressed in Escherichia coli, purified and characterized. The enzyme is optimally active at 90 °C and pH 9.0 with a specific activity of 2164 U/mg towards L-asparagine. Kinetic parameters K(M) and V(max) for the enzyme are 2.8 mM and 1200 µM/min, respectively. TsA is stable in urea solutions 0–6 M and displays no significant changes of the activity in the presence of metal ions Ni(2+), Cu(2+), Mg(2+), Zn(2+) and Ca(2+) and EDTA added in concentrations 1 and 10 mmol/L except for Fe(3+). The enzyme retains 86% of its initial activity after 20 min incubation at 90 °C, which should be enough to reduce acrylamide formation in foods processed at elevated temperatures. TsA displays strong cytotoxic activity toward cancer cell lines K562, A549 and Sk-Br-3, while normal human fibroblasts WI-38 are almost unsensitive to it. The enzyme seems to be a promising candidate for further investigation and biotechnology application. |
format | Online Article Text |
id | pubmed-8470970 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-84709702021-09-27 A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application Dumina, Maria Zhgun, Alexander Pokrovskaya, Marina Aleksandrova, Svetlana Zhdanov, Dmitry Sokolov, Nikolay El’darov, Michael Int J Mol Sci Article L-asparaginase (L-ASNase) is a vital enzyme with a broad range of applications in medicine and food industry. Drawbacks of current commercial L-ASNases stimulate the search for better-producing sources of the enzyme, and extremophiles are especially attractive in this view. In this study, a novel L-asparaginase originating from the hyperthermophilic archaeon Thermococcus sibiricus (TsA) was expressed in Escherichia coli, purified and characterized. The enzyme is optimally active at 90 °C and pH 9.0 with a specific activity of 2164 U/mg towards L-asparagine. Kinetic parameters K(M) and V(max) for the enzyme are 2.8 mM and 1200 µM/min, respectively. TsA is stable in urea solutions 0–6 M and displays no significant changes of the activity in the presence of metal ions Ni(2+), Cu(2+), Mg(2+), Zn(2+) and Ca(2+) and EDTA added in concentrations 1 and 10 mmol/L except for Fe(3+). The enzyme retains 86% of its initial activity after 20 min incubation at 90 °C, which should be enough to reduce acrylamide formation in foods processed at elevated temperatures. TsA displays strong cytotoxic activity toward cancer cell lines K562, A549 and Sk-Br-3, while normal human fibroblasts WI-38 are almost unsensitive to it. The enzyme seems to be a promising candidate for further investigation and biotechnology application. MDPI 2021-09-13 /pmc/articles/PMC8470970/ /pubmed/34576056 http://dx.doi.org/10.3390/ijms22189894 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Dumina, Maria Zhgun, Alexander Pokrovskaya, Marina Aleksandrova, Svetlana Zhdanov, Dmitry Sokolov, Nikolay El’darov, Michael A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title | A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title_full | A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title_fullStr | A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title_full_unstemmed | A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title_short | A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application |
title_sort | novel l-asparaginase from hyperthermophilic archaeon thermococcus sibiricus: heterologous expression and characterization for biotechnology application |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470970/ https://www.ncbi.nlm.nih.gov/pubmed/34576056 http://dx.doi.org/10.3390/ijms22189894 |
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