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Title: Proteolysis of micellar β-casein by trypsin: secondary structure characterization and kinetic modeling at different enzyme concentrations
Authors: Vorob’ev, Mikhail M.
Açıkgöz, Burçin Dersu
Güler, Günnur
Golovanov, Andrey V.
Sinitsyna, Olga V.
Keywords: Atomic force microscopy
Fourier-transform infrared spectroscopy
Proteolysis kinetics
Publisher: MDPI
Abstract: Tryptic proteolysis of protein micelles was studied using β-casein (β-CN) as an example. Hydrolysis of specific peptide bonds in β-CN leads to the degradation and rearrangement of the original micelles and the formation of new nanoparticles from their fragments. Samples of these nanoparticles dried on a mica surface were characterized by atomic force microscopy (AFM) when the proteolytic reaction had been stopped by tryptic inhibitor or by heating. The changes in the content of β-sheets, α-helices, and hydrolysis products during proteolysis were estimated by using Fourier-transform infrared (FTIR) spectroscopy. In the current study, a simple kinetic model with three successive stages is proposed to predict the rearrangement of nanoparticles and the formation of proteolysis products, as well as changes in the secondary structure during proteolysis at various enzyme concentrations. The model determines for which steps the rate constants are proportional to the enzyme concentration, and in which intermediate nano-components the protein secondary structure is retained and in which it is reduced. The model predictions were in agreement with the FTIR results for tryptic hydrolysis of β-CN at different concentrations of the enzyme.
Description: This work was supported by both RFBR (№ 20-53-46006 to M.M. Vorobev) and TÜBİTAK-2532 (№ 119N423 to G. Güler). The work of the Russian team was supported by the Ministry of Science and Higher Education of the Russian Federation (Contract No. 075-03-2023-642).
ISSN: 1661-6596
Appears in Collections:Physics / Fizik
PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

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