Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI)
Lipase B from Candida antarctica (CALB) was immobilized on octyl agarose (OC) and physically modified with polyethyleneimine (PEI) in order to confer a strong ion exchange character to the enzyme and thus enable the immobilization of other enzymes on its surface. The enzyme activity was fully mainta...
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MDPI AG
2016-06-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/21/6/751 |
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| author | Sara Peirce Veymar G. Tacias-Pascacio Maria Elena Russo Antonio Marzocchella José J. Virgen-Ortíz Roberto Fernandez-Lafuente |
| author_facet | Sara Peirce Veymar G. Tacias-Pascacio Maria Elena Russo Antonio Marzocchella José J. Virgen-Ortíz Roberto Fernandez-Lafuente |
| author_sort | Sara Peirce |
| collection | DOAJ |
| description | Lipase B from Candida antarctica (CALB) was immobilized on octyl agarose (OC) and physically modified with polyethyleneimine (PEI) in order to confer a strong ion exchange character to the enzyme and thus enable the immobilization of other enzymes on its surface. The enzyme activity was fully maintained during the coating and the thermal stability was marginally improved. The enzyme release from the support by incubation in the non-ionic detergent Triton X-100 was more difficult after the PEI-coating, suggesting that some intermolecular physical crosslinking had occurred, making this desorption more difficult. Thermal stability was marginally improved, but the stability of the OCCALB-PEI was significantly better than that of OCCALB during inactivation in mixtures of aqueous buffer and organic cosolvents. SDS-PAGE analysis of the inactivated biocatalyst showed the OCCALB released some enzyme to the medium during inactivation, and this was partially prevented by coating with PEI. This effect was obtained without preventing the possibility of reuse of the support by incubation in 2% ionic detergents. That way, this modified CALB not only has a strong anion exchange nature, while maintaining the activity, but it also shows improved stability under diverse reaction conditions without affecting the reversibility of the immobilization. |
| first_indexed | 2024-12-12T16:38:37Z |
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| language | English |
| last_indexed | 2024-12-12T16:38:37Z |
| publishDate | 2016-06-01 |
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| spelling | doaj.art-2e5d3b4aeebc4a7797adfc55fc8861962022-12-22T00:18:36ZengMDPI AGMolecules1420-30492016-06-0121675110.3390/molecules21060751molecules21060751Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI)Sara Peirce0Veymar G. Tacias-Pascacio1Maria Elena Russo2Antonio Marzocchella3José J. Virgen-Ortíz4Roberto Fernandez-Lafuente5Departamento de Biocatálisis, Instituto de Catálisis-CSIC, C/Marie Curie 2, Campus UAM-CSIC Cantoblanco, 28049 Madrid, SpainDepartamento de Biocatálisis, Instituto de Catálisis-CSIC, C/Marie Curie 2, Campus UAM-CSIC Cantoblanco, 28049 Madrid, SpainIstituto di Ricerche sulla Combustione—Consiglio Nazionale delle Ricerche, 80125 Napoli, ItalyDipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale, Universita’ degli Studi di Napoli Federico II, 80125 Napoli, ItalyDepartamento de Biocatálisis, Instituto de Catálisis-CSIC, C/Marie Curie 2, Campus UAM-CSIC Cantoblanco, 28049 Madrid, SpainDepartamento de Biocatálisis, Instituto de Catálisis-CSIC, C/Marie Curie 2, Campus UAM-CSIC Cantoblanco, 28049 Madrid, SpainLipase B from Candida antarctica (CALB) was immobilized on octyl agarose (OC) and physically modified with polyethyleneimine (PEI) in order to confer a strong ion exchange character to the enzyme and thus enable the immobilization of other enzymes on its surface. The enzyme activity was fully maintained during the coating and the thermal stability was marginally improved. The enzyme release from the support by incubation in the non-ionic detergent Triton X-100 was more difficult after the PEI-coating, suggesting that some intermolecular physical crosslinking had occurred, making this desorption more difficult. Thermal stability was marginally improved, but the stability of the OCCALB-PEI was significantly better than that of OCCALB during inactivation in mixtures of aqueous buffer and organic cosolvents. SDS-PAGE analysis of the inactivated biocatalyst showed the OCCALB released some enzyme to the medium during inactivation, and this was partially prevented by coating with PEI. This effect was obtained without preventing the possibility of reuse of the support by incubation in 2% ionic detergents. That way, this modified CALB not only has a strong anion exchange nature, while maintaining the activity, but it also shows improved stability under diverse reaction conditions without affecting the reversibility of the immobilization.http://www.mdpi.com/1420-3049/21/6/751reversible immobilizationinterfacial adsorptionPEI modificationenzyme stabilizationenzyme physical intermolecular crosslinking |
| spellingShingle | Sara Peirce Veymar G. Tacias-Pascacio Maria Elena Russo Antonio Marzocchella José J. Virgen-Ortíz Roberto Fernandez-Lafuente Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) Molecules reversible immobilization interfacial adsorption PEI modification enzyme stabilization enzyme physical intermolecular crosslinking |
| title | Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) |
| title_full | Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) |
| title_fullStr | Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) |
| title_full_unstemmed | Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) |
| title_short | Stabilization of Candida antarctica Lipase B (CALB) Immobilized on Octyl Agarose by Treatment with Polyethyleneimine (PEI) |
| title_sort | stabilization of candida antarctica lipase b calb immobilized on octyl agarose by treatment with polyethyleneimine pei |
| topic | reversible immobilization interfacial adsorption PEI modification enzyme stabilization enzyme physical intermolecular crosslinking |
| url | http://www.mdpi.com/1420-3049/21/6/751 |
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