Glutaraldehyde modification of lipases immobilized on octyl agarose beads: Roles of the support enzyme loading and chemical amination of the enzyme on the final enzyme features
Lipase B from Candida antarctica (CALB) and lipase from Thermomyces lanuginosus (TLL) have been immobilized on octyl agarose at low loading and at a loading exceeding the maximum support capacity. Then, the enzymes have been treated with glutaraldehyde and inactivated at pH 7.0 in Tris-HCl, sodium p...
| Autores: | , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/356213 |
| Acceso en línea: | http://hdl.handle.net/10261/356213 https://www.scopus.com/inward/record.uri?eid=2-s2.0-85165539924&doi=10.1016%2fj.ijbiomac.2023.125853&partnerID=40&md5=863bd0274ebd7efed524e837d339289d |
| Access Level: | acceso abierto |
| Palabra clave: | Enzyme stabilization Inter and intramolecular enzyme crosslinking Effect of enzyme loading Effect of the inactivating buffer on enzyme stabilityactivating buffer on enzyme stability Amination Candida Enzymes Immobilized Fungal Proteins Glutaral Lipases Sepharose Amines Chemical modification Crosslinking Loading Sodium compounds Agarose Glutaraldehydes Octyl agarose bead triacylglycerol lipase unclassified drug immobilized enzyme sepharose Enzyme crosslinking Enzyme loading biocatalyst Controlled study pH polyacrylamide gel electrophoresis Thermomyces lanuginosus Chemistry Stabilization |
| Sumario: | Lipase B from Candida antarctica (CALB) and lipase from Thermomyces lanuginosus (TLL) have been immobilized on octyl agarose at low loading and at a loading exceeding the maximum support capacity. Then, the enzymes have been treated with glutaraldehyde and inactivated at pH 7.0 in Tris-HCl, sodium phosphate and HEPES, giving different stabilities. Stabilization (depending on the buffer) of the highly loaded biocatalysts was found, very likely as a consequence of the detected intermolecular crosslinkings. This did not occur for the lowly loaded biocatalysts. Next, the enzymes were chemically aminated and then treated with glutaraldehyde. In the case of TLL, the intramolecular crosslinkings (visible by the apparent reduction of the protein size) increased enzyme stability of the lowly loaded biocatalysts, an effect that was further increased for the highly loaded biocatalysts due to intermolecular crosslinkings. Using CALB, the intramolecular crosslinkings were less intense, and the stabilization was lower, even though the intermolecular crosslinkings were quite intense for the highly loaded biocatalyst. The stabilization detected depended on the inactivation buffer. The interactions between enzyme loading and inactivating buffer on the effects of the chemical modifications suggest that the modification and inactivation studies must be performed under the target biocatalysts and conditions. © 2023 The Authors |
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