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Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency
Enzyme immobilization on adequate carriers is a challenging strategy. Understanding the enzyme‐carrier interactions and their effects on enzyme conformation and bioactivity is critical. In this study, a meso‐macropores silica (MMS) was used to immobilize β‐galactosidase from the yeast Kluyveromyces...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10369856/ https://www.ncbi.nlm.nih.gov/pubmed/36515233 http://dx.doi.org/10.1002/cplu.202200340 |
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author | Videira‐Quintela, Diogo Guillén, Francisco Prazeres, Sofia F. Montalvo, Gemma |
author_facet | Videira‐Quintela, Diogo Guillén, Francisco Prazeres, Sofia F. Montalvo, Gemma |
author_sort | Videira‐Quintela, Diogo |
collection | PubMed |
description | Enzyme immobilization on adequate carriers is a challenging strategy. Understanding the enzyme‐carrier interactions and their effects on enzyme conformation and bioactivity is critical. In this study, a meso‐macropores silica (MMS) was used to immobilize β‐galactosidase from the yeast Kluyveromyces lactis (β‐gal‐KL) by physical adsorption. The bioactivity of the immobilized β‐gal‐KL was altered, evidenced by the increased K(m), decreased V(max) and k(cat), and increased activity at alkaline values. By performing infrared spectroscopy analysis and subsequent secondary structure assessment from the amide I band, the immobilized β‐gal‐KL suffered a β‐sheet (∼31–35 %) to α‐helix (∼15–19 %) transition with increased turns (∼21–22 %) with respect to the free β‐gal‐KL having ∼12 % α‐helix, ∼42 % β‐sheet, and ∼17 % turns. These findings led us to correlate the observed bioactivity performance to structural alterations to a non‐native conformation. The presented line of thought can lead to a better understanding of the reasons causing bioactivity alterations upon enzyme immobilization. |
format | Online Article Text |
id | pubmed-10369856 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-103698562023-07-27 Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency Videira‐Quintela, Diogo Guillén, Francisco Prazeres, Sofia F. Montalvo, Gemma Chempluschem Research Articles Enzyme immobilization on adequate carriers is a challenging strategy. Understanding the enzyme‐carrier interactions and their effects on enzyme conformation and bioactivity is critical. In this study, a meso‐macropores silica (MMS) was used to immobilize β‐galactosidase from the yeast Kluyveromyces lactis (β‐gal‐KL) by physical adsorption. The bioactivity of the immobilized β‐gal‐KL was altered, evidenced by the increased K(m), decreased V(max) and k(cat), and increased activity at alkaline values. By performing infrared spectroscopy analysis and subsequent secondary structure assessment from the amide I band, the immobilized β‐gal‐KL suffered a β‐sheet (∼31–35 %) to α‐helix (∼15–19 %) transition with increased turns (∼21–22 %) with respect to the free β‐gal‐KL having ∼12 % α‐helix, ∼42 % β‐sheet, and ∼17 % turns. These findings led us to correlate the observed bioactivity performance to structural alterations to a non‐native conformation. The presented line of thought can lead to a better understanding of the reasons causing bioactivity alterations upon enzyme immobilization. John Wiley and Sons Inc. 2022-12-14 2022-12 /pmc/articles/PMC10369856/ /pubmed/36515233 http://dx.doi.org/10.1002/cplu.202200340 Text en © 2022 The Authors. ChemPlusChem published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Videira‐Quintela, Diogo Guillén, Francisco Prazeres, Sofia F. Montalvo, Gemma Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title | Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title_full | Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title_fullStr | Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title_full_unstemmed | Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title_short | Immobilization of Kluyveromyces lactis β‐Galactosidase on Meso‐macroporous Silica: Use of Infrared Spectroscopy to Rationalize the Catalytic Efficiency |
title_sort | immobilization of kluyveromyces lactis β‐galactosidase on meso‐macroporous silica: use of infrared spectroscopy to rationalize the catalytic efficiency |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10369856/ https://www.ncbi.nlm.nih.gov/pubmed/36515233 http://dx.doi.org/10.1002/cplu.202200340 |
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