Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods
Thermal stability of lactase (β-galactosidase) enzyme has been studied by a variety of physico-chemical methods. β-galactosidase is the main active ingredient of medications for lactose intolerance. It is typically produced industrially by the Aspergillus oryzae filamentous fungus. Lactase was used...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | Biotechnology Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X21000539 |
| _version_ | 1818864760508121088 |
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| author | Márton Király Borbála Dalmadi Kiss Péter Horváth László Drahos Arash Mirzahosseini Gyula Pálfy István Antal Krisztina Ludányi |
| author_facet | Márton Király Borbála Dalmadi Kiss Péter Horváth László Drahos Arash Mirzahosseini Gyula Pálfy István Antal Krisztina Ludányi |
| author_sort | Márton Király |
| collection | DOAJ |
| description | Thermal stability of lactase (β-galactosidase) enzyme has been studied by a variety of physico-chemical methods. β-galactosidase is the main active ingredient of medications for lactose intolerance. It is typically produced industrially by the Aspergillus oryzae filamentous fungus. Lactase was used as a model to help understand thermal stability of enzyme-type biopharmaceuticals. Enzyme activity (hydrolyzation of lactose) of β-galactosidase was determined after storing the solid enzyme substance at various temperatures. For a better understanding of the relationship between structure and activity changes we determined the mass and size of the molecules with gel electrophoresis and dynamic light scattering and detected aggregation processes. A bottom-up proteomic procedure was used to determine the primary amino acid sequence and to investigate changes in the N-glycosylation pattern of the protein. NMR and CD spectroscopic methods were used to observe changes in higher order structures and to reveal relationships between structural and functional changes. |
| first_indexed | 2024-12-19T10:36:47Z |
| format | Article |
| id | doaj.art-41ae5c5dae78436196a873ec46b7a4bf |
| institution | Directory Open Access Journal |
| issn | 2215-017X |
| language | English |
| last_indexed | 2024-12-19T10:36:47Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj.art-41ae5c5dae78436196a873ec46b7a4bf2022-12-21T20:25:35ZengElsevierBiotechnology Reports2215-017X2021-06-0130e00637Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methodsMárton Király0Borbála Dalmadi Kiss1Péter Horváth2László Drahos3Arash Mirzahosseini4Gyula Pálfy5István Antal6Krisztina Ludányi7Department of Pharmaceutics, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, HungaryDepartment of Pharmaceutics, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, HungaryDepartment of Pharmaceutical Chemistry, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, HungaryMS Proteomics Research Group, Research Centre for Natural Sciences, Magyar Tudósok körútja 2., H-1117, Budapest, HungaryDepartment of Pharmaceutical Chemistry, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, HungaryLaboratory of Structural Chemistry and Biology, Institute of Chemistry, Eötvös Loránd University, Pázmány P. sétány 1/A, 1117, Budapest, Hungary; Protein Modeling Group HAS-ELTE, Institute of Chemistry, Eötvös Loránd University, 1538, Budapest, P.O.B. 32, HungaryDepartment of Pharmaceutics, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, HungaryDepartment of Pharmaceutics, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, Hungary; Corresponding author.Thermal stability of lactase (β-galactosidase) enzyme has been studied by a variety of physico-chemical methods. β-galactosidase is the main active ingredient of medications for lactose intolerance. It is typically produced industrially by the Aspergillus oryzae filamentous fungus. Lactase was used as a model to help understand thermal stability of enzyme-type biopharmaceuticals. Enzyme activity (hydrolyzation of lactose) of β-galactosidase was determined after storing the solid enzyme substance at various temperatures. For a better understanding of the relationship between structure and activity changes we determined the mass and size of the molecules with gel electrophoresis and dynamic light scattering and detected aggregation processes. A bottom-up proteomic procedure was used to determine the primary amino acid sequence and to investigate changes in the N-glycosylation pattern of the protein. NMR and CD spectroscopic methods were used to observe changes in higher order structures and to reveal relationships between structural and functional changes.http://www.sciencedirect.com/science/article/pii/S2215017X21000539NMRProteomicsβ-GalactosidaseAggregationMass spectrometry |
| spellingShingle | Márton Király Borbála Dalmadi Kiss Péter Horváth László Drahos Arash Mirzahosseini Gyula Pálfy István Antal Krisztina Ludányi Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods Biotechnology Reports NMR Proteomics β-Galactosidase Aggregation Mass spectrometry |
| title | Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| title_full | Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| title_fullStr | Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| title_full_unstemmed | Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| title_short | Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| title_sort | investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods |
| topic | NMR Proteomics β-Galactosidase Aggregation Mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X21000539 |
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