Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins
Different materials containing carboxylic groups have been functionalized with geranyl-amine molecules by using an EDC/NHS strategy. Chemical modification of the support was confirmed by XRD, UV-spectrophotometer, and FT-IR. This geranyl-functionalized material was successfully applied for four diff...
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| Format: | Article |
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MDPI AG
2021-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/10/3028 |
| _version_ | 1797533467974565888 |
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| author | Jana Brabcova Alicia Andreu David Aguilera Zaida Cabrera Blanca de las Rivas Rosario Muñoz Jose M. Palomo |
| author_facet | Jana Brabcova Alicia Andreu David Aguilera Zaida Cabrera Blanca de las Rivas Rosario Muñoz Jose M. Palomo |
| author_sort | Jana Brabcova |
| collection | DOAJ |
| description | Different materials containing carboxylic groups have been functionalized with geranyl-amine molecules by using an EDC/NHS strategy. Chemical modification of the support was confirmed by XRD, UV-spectrophotometer, and FT-IR. This geranyl-functionalized material was successfully applied for four different strategies of site-selective immobilization of proteins at room temperature and aqueous media. A reversible hydrophobic immobilization of proteins (lipases, phosphoglucosidases, or tyrosinase) was performed in neutral pH in yields from 40 to >99%. An increase of the activity in the case of lipases was observed from a range of 2 to 4 times with respect to the initial activity in solution. When chemically or genetically functionalized cysteine enzymes were used, the covalent immobilization, via a selective thiol-alkene reaction, was observed in the presence of geranyl support at pH 8 in lipases in the presence of detergent (to avoid the previous hydrophobic interactions). Covalent attachment was confirmed with no release of protein after immobilization by incubation with hydrophobic molecules. In the case of a selenium-containing enzyme produced by the selenomethionine pathway, the selective immobilization was successfully yielded at acidic pH (pH 5) (89%) much better than at pH 8. In addition, when an azido-enzyme was produced by the azide–homoalanine pathway, the selective immobilization was successful at pH 6 and in the presence of CuI for the click chemistry reaction. |
| first_indexed | 2024-03-10T11:15:54Z |
| format | Article |
| id | doaj.art-2d1d31cf5cac472aa66ca0fbf5afbe6f |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T11:15:54Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-2d1d31cf5cac472aa66ca0fbf5afbe6f2023-11-21T20:25:05ZengMDPI AGMolecules1420-30492021-05-012610302810.3390/molecules26103028Geranyl Functionalized Materials for Site-Specific Co-Immobilization of ProteinsJana Brabcova0Alicia Andreu1David Aguilera2Zaida Cabrera3Blanca de las Rivas4Rosario Muñoz5Jose M. Palomo6Department of Biocatalysis, Institute of Catalysis (CSIC), Marie Curie 2, 28049 Madrid, SpainDepartment of Biocatalysis, Institute of Catalysis (CSIC), Marie Curie 2, 28049 Madrid, SpainDepartment of Biocatalysis, Institute of Catalysis (CSIC), Marie Curie 2, 28049 Madrid, SpainSchool of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Avda, Brasil, 2085 Valparaíso, ChileBacterial Biotechnology, Institute of Food Science, Technology and Nutrition (ICTAN-CSIC), Juan de la Cierva 3, 28006 Madrid, SpainBacterial Biotechnology, Institute of Food Science, Technology and Nutrition (ICTAN-CSIC), Juan de la Cierva 3, 28006 Madrid, SpainDepartment of Biocatalysis, Institute of Catalysis (CSIC), Marie Curie 2, 28049 Madrid, SpainDifferent materials containing carboxylic groups have been functionalized with geranyl-amine molecules by using an EDC/NHS strategy. Chemical modification of the support was confirmed by XRD, UV-spectrophotometer, and FT-IR. This geranyl-functionalized material was successfully applied for four different strategies of site-selective immobilization of proteins at room temperature and aqueous media. A reversible hydrophobic immobilization of proteins (lipases, phosphoglucosidases, or tyrosinase) was performed in neutral pH in yields from 40 to >99%. An increase of the activity in the case of lipases was observed from a range of 2 to 4 times with respect to the initial activity in solution. When chemically or genetically functionalized cysteine enzymes were used, the covalent immobilization, via a selective thiol-alkene reaction, was observed in the presence of geranyl support at pH 8 in lipases in the presence of detergent (to avoid the previous hydrophobic interactions). Covalent attachment was confirmed with no release of protein after immobilization by incubation with hydrophobic molecules. In the case of a selenium-containing enzyme produced by the selenomethionine pathway, the selective immobilization was successfully yielded at acidic pH (pH 5) (89%) much better than at pH 8. In addition, when an azido-enzyme was produced by the azide–homoalanine pathway, the selective immobilization was successful at pH 6 and in the presence of CuI for the click chemistry reaction.https://www.mdpi.com/1420-3049/26/10/3028geranylfunctionalized materialshydrophobic interactionsthiol-alkene reactionseleno-alkene reactionazide cycloaddition |
| spellingShingle | Jana Brabcova Alicia Andreu David Aguilera Zaida Cabrera Blanca de las Rivas Rosario Muñoz Jose M. Palomo Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins Molecules geranyl functionalized materials hydrophobic interactions thiol-alkene reaction seleno-alkene reaction azide cycloaddition |
| title | Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins |
| title_full | Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins |
| title_fullStr | Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins |
| title_full_unstemmed | Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins |
| title_short | Geranyl Functionalized Materials for Site-Specific Co-Immobilization of Proteins |
| title_sort | geranyl functionalized materials for site specific co immobilization of proteins |
| topic | geranyl functionalized materials hydrophobic interactions thiol-alkene reaction seleno-alkene reaction azide cycloaddition |
| url | https://www.mdpi.com/1420-3049/26/10/3028 |
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