Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i>
Neuraminidase (NA), as an important protein of influenza virus, represents a promising target for the development of new antiviral agents for the treatment and prevention of influenza A and B. Bacterial host strain <i>Escherichia coli</i> BL21 (DE3)pLysS containing the NA gene of the H1N...
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MDPI AG
2022-02-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/2/331 |
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| author | Sabina Lipničanová Barbora Legerská Daniela Chmelová Miroslav Ondrejovič Stanislav Miertuš |
| author_facet | Sabina Lipničanová Barbora Legerská Daniela Chmelová Miroslav Ondrejovič Stanislav Miertuš |
| author_sort | Sabina Lipničanová |
| collection | DOAJ |
| description | Neuraminidase (NA), as an important protein of influenza virus, represents a promising target for the development of new antiviral agents for the treatment and prevention of influenza A and B. Bacterial host strain <i>Escherichia coli</i> BL21 (DE3)pLysS containing the NA gene of the H1N1 influenza virus produced this overexpressed enzyme in the insoluble fraction of cells in the form of inclusion bodies. The aim of this work was to investigate the effect of independent variables (propagation time, isopropyl <i>β</i>-<span style="font-variant: small-caps;">d</span>-1-thiogalactopyranoside (IPTG) concentration and expression time) on NA accumulation in inclusion bodies and to optimize these conditions by response surface methodology (RSM). The maximum yield of NA (112.97 ± 2.82 U/g) was achieved under optimal conditions, namely, a propagation time of 7.72 h, IPTG concentration of 1.82 mM and gene expression time of 7.35 h. This study demonstrated that bacterially expressed NA was enzymatically active. |
| first_indexed | 2024-03-09T22:29:09Z |
| format | Article |
| id | doaj.art-a49593c09041477d8c60237e697ae783 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T22:29:09Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-a49593c09041477d8c60237e697ae7832023-11-23T19:00:13ZengMDPI AGBiomolecules2218-273X2022-02-0112233110.3390/biom12020331Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i>Sabina Lipničanová0Barbora Legerská1Daniela Chmelová2Miroslav Ondrejovič3Stanislav Miertuš4Department of Biotechnology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, J. Herdu 2, SK-91701 Trnava, SlovakiaDepartment of Biotechnology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, J. Herdu 2, SK-91701 Trnava, SlovakiaDepartment of Biotechnology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, J. Herdu 2, SK-91701 Trnava, SlovakiaDepartment of Biotechnology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, J. Herdu 2, SK-91701 Trnava, SlovakiaDepartment of Biotechnology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, J. Herdu 2, SK-91701 Trnava, SlovakiaNeuraminidase (NA), as an important protein of influenza virus, represents a promising target for the development of new antiviral agents for the treatment and prevention of influenza A and B. Bacterial host strain <i>Escherichia coli</i> BL21 (DE3)pLysS containing the NA gene of the H1N1 influenza virus produced this overexpressed enzyme in the insoluble fraction of cells in the form of inclusion bodies. The aim of this work was to investigate the effect of independent variables (propagation time, isopropyl <i>β</i>-<span style="font-variant: small-caps;">d</span>-1-thiogalactopyranoside (IPTG) concentration and expression time) on NA accumulation in inclusion bodies and to optimize these conditions by response surface methodology (RSM). The maximum yield of NA (112.97 ± 2.82 U/g) was achieved under optimal conditions, namely, a propagation time of 7.72 h, IPTG concentration of 1.82 mM and gene expression time of 7.35 h. This study demonstrated that bacterially expressed NA was enzymatically active.https://www.mdpi.com/2218-273X/12/2/331neuraminidaseinclusion bodyrecombinant proteinexpressionoptimizationresponse surface methodology |
| spellingShingle | Sabina Lipničanová Barbora Legerská Daniela Chmelová Miroslav Ondrejovič Stanislav Miertuš Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> Biomolecules neuraminidase inclusion body recombinant protein expression optimization response surface methodology |
| title | Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> |
| title_full | Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> |
| title_fullStr | Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> |
| title_full_unstemmed | Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> |
| title_short | Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in <i>Escherichia coli</i> |
| title_sort | optimization of an inclusion body based production of the influenza virus neuraminidase in i escherichia coli i |
| topic | neuraminidase inclusion body recombinant protein expression optimization response surface methodology |
| url | https://www.mdpi.com/2218-273X/12/2/331 |
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