Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids
High quality nucleic acids (with high integrity, purity, and biological activity) have become indispensable products of modern society, both in molecular diagnosis and to be used as biopharmaceuticals. As the current methods available for the extraction and purification of nucleic acids are laboriou...
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| Format: | Article |
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MDPI AG
2021-10-01
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| Online Access: | https://www.mdpi.com/2075-1729/11/10/1090 |
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| author | Patrícia Pereira Augusto Q. Pedro Márcia C. Neves João C. Martins Inês Rodrigues Mara G. Freire Fani Sousa |
| author_facet | Patrícia Pereira Augusto Q. Pedro Márcia C. Neves João C. Martins Inês Rodrigues Mara G. Freire Fani Sousa |
| author_sort | Patrícia Pereira |
| collection | DOAJ |
| description | High quality nucleic acids (with high integrity, purity, and biological activity) have become indispensable products of modern society, both in molecular diagnosis and to be used as biopharmaceuticals. As the current methods available for the extraction and purification of nucleic acids are laborious, time-consuming, and usually rely on the use of hazardous chemicals, there is an unmet need towards the development of more sustainable and cost-effective technologies for nucleic acids purification. Accordingly, this study addresses the preparation and evaluation of silica-based materials chemically modified with chloride-based ionic liquids (supported ionic liquids, SILs) as potential materials to effectively isolate RNAs. The investigated chloride-based SILs comprise the following cations: 1-methyl-3-propylimidazolium, triethylpropylammonium, dimethylbutylpropylammonium, and trioctylpropylammonium. All SILs were synthesized by us and characterized by solid-state <sup>13</sup>C Nuclear Magnetic Resonance (NMR), Scanning Electron Microscopy (SEM), elemental analysis, and zeta potential measurements, confirming the successful covalent attachment of each IL cation with no relevant changes in the morphology of materials. Their innovative application as chromatographic supports for the isolation of recombinant RNA was then evaluated. Adsorption kinetics of transfer RNA (tRNA) on the modified silica-based materials were investigated at 25 °C. Irrespective to the immobilized IL, the adsorption experimental data are better described by a pseudo first-order model, and maximum tRNA binding capacities of circa 16 µmol of tRNA/g of material were achieved with silica modified with 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Furthermore, the multimodal character displayed by SILs was explored towards the purification of tRNA from <i>Escherichia coli</i> lysates, which in addition to tRNA contain ribosomal RNA and genomic DNA. The best performance on the tRNA isolation was achieved with SILs comprising 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Overall, the IL modified silica-based materials represent a more efficient, sustainable, and cost-effective technology for the purification of bacterial RNAs, paving the way for their use in the purification of distinct biomolecules or nucleic acids from other sources. |
| first_indexed | 2024-03-10T06:26:46Z |
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| id | doaj.art-41a10273c87043d692b22f03f87c6e34 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-10T06:26:46Z |
| publishDate | 2021-10-01 |
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| record_format | Article |
| series | Life |
| spelling | doaj.art-41a10273c87043d692b22f03f87c6e342023-11-22T18:53:30ZengMDPI AGLife2075-17292021-10-011110109010.3390/life11101090Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic LiquidsPatrícia Pereira0Augusto Q. Pedro1Márcia C. Neves2João C. Martins3Inês Rodrigues4Mara G. Freire5Fani Sousa6CEMMPRE, Department of Chemical Engineering, University of Coimbra, Rua Sílvio Lima-Pólo II, 3030-790 Coimbra, PortugalCICECO—Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, PortugalCICECO—Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, PortugalCICS-UBI—Health Sciences Research Centre, Universidade da Beira Interior, 6200-506 Covilhã, PortugalCICS-UBI—Health Sciences Research Centre, Universidade da Beira Interior, 6200-506 Covilhã, PortugalCICECO—Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, PortugalCICS-UBI—Health Sciences Research Centre, Universidade da Beira Interior, 6200-506 Covilhã, PortugalHigh quality nucleic acids (with high integrity, purity, and biological activity) have become indispensable products of modern society, both in molecular diagnosis and to be used as biopharmaceuticals. As the current methods available for the extraction and purification of nucleic acids are laborious, time-consuming, and usually rely on the use of hazardous chemicals, there is an unmet need towards the development of more sustainable and cost-effective technologies for nucleic acids purification. Accordingly, this study addresses the preparation and evaluation of silica-based materials chemically modified with chloride-based ionic liquids (supported ionic liquids, SILs) as potential materials to effectively isolate RNAs. The investigated chloride-based SILs comprise the following cations: 1-methyl-3-propylimidazolium, triethylpropylammonium, dimethylbutylpropylammonium, and trioctylpropylammonium. All SILs were synthesized by us and characterized by solid-state <sup>13</sup>C Nuclear Magnetic Resonance (NMR), Scanning Electron Microscopy (SEM), elemental analysis, and zeta potential measurements, confirming the successful covalent attachment of each IL cation with no relevant changes in the morphology of materials. Their innovative application as chromatographic supports for the isolation of recombinant RNA was then evaluated. Adsorption kinetics of transfer RNA (tRNA) on the modified silica-based materials were investigated at 25 °C. Irrespective to the immobilized IL, the adsorption experimental data are better described by a pseudo first-order model, and maximum tRNA binding capacities of circa 16 µmol of tRNA/g of material were achieved with silica modified with 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Furthermore, the multimodal character displayed by SILs was explored towards the purification of tRNA from <i>Escherichia coli</i> lysates, which in addition to tRNA contain ribosomal RNA and genomic DNA. The best performance on the tRNA isolation was achieved with SILs comprising 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Overall, the IL modified silica-based materials represent a more efficient, sustainable, and cost-effective technology for the purification of bacterial RNAs, paving the way for their use in the purification of distinct biomolecules or nucleic acids from other sources.https://www.mdpi.com/2075-1729/11/10/1090adsorption kineticsdownstream processesliquid chromatographyRNAsilicasupported ionic liquids |
| spellingShingle | Patrícia Pereira Augusto Q. Pedro Márcia C. Neves João C. Martins Inês Rodrigues Mara G. Freire Fani Sousa Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids Life adsorption kinetics downstream processes liquid chromatography RNA silica supported ionic liquids |
| title | Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids |
| title_full | Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids |
| title_fullStr | Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids |
| title_full_unstemmed | Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids |
| title_short | Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids |
| title_sort | efficient isolation of bacterial rnas using silica based materials modified with ionic liquids |
| topic | adsorption kinetics downstream processes liquid chromatography RNA silica supported ionic liquids |
| url | https://www.mdpi.com/2075-1729/11/10/1090 |
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