Investigating thermal stability based on the structural changes of lactase enzyme by several orthogonal methods

Biotechnology Reports

卷:30

István Antal¹  Arash Mirzahosseini²  Péter Horváth²  Krisztina Ludányi³  Márton Király³  Borbála Dalmadi Kiss³  Gyula Pálfy⁴  László Drahos⁵ 

[1] Protein Modeling Group HAS-ELTE, Institute of Chemistry, Eötvös Loránd University, 1538, Budapest, P.O.B. 32, Hungary;

[2] Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, Hungary;

[3] Department of Pharmaceutics, Faculty of Pharmacy, Semmelweis University, Hőgyes Endre u. 7., 1092, Budapest, Hungary;

[4] Laboratory of Structural Chemistry and Biology, Institute of Chemistry, Eötvös Loránd University, Pázmány P. sétány 1/A, 1117, Budapest, Hungary;

[5] MS Proteomics Research Group, Research Centre for Natural Sciences, Magyar Tudósok körútja 2., H-1117, Budapest, Hungary;

关键词: NMR;    Proteomics;    β-Galactosidase;    Aggregation;    Mass spectrometry;   

DOI  :  

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown