Tryptic hydrolysis of β-lactoglobulin: A generic approach to describe the hydrolysis kinetic and release of peptides

Leeb, Elena; Stefan, Thorsten; Letzel, Thomas; Hinrichs, Jörg; Kulozik, Ulrich

doi:10.1016/j.idairyj.2020.104666

Cited by 6 publications

(1 citation statement)

References 30 publications

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“…These studies made it possible to identify the main intermediate and final peptide fragments for proteolysis of β-LG and β-CN by trypsin. The peptide fragments of β-LG analyzed at various times of proteolysis were used to establish the sequence of fragmentation steps [ 24 , 33 , 34 ], and a system of differential equations was derived for describing these steps [ 33 ]. The relatively low concentrations of the protein substrates, such as 0.25 g/L in this work, guarantee the absence of the reverse reaction of proteolysis, the plastein reaction [ 35 ], leading to the formation of new peptide bonds.…”

Section: Introductionmentioning

confidence: 99%

Modeling of Proteolysis of β-Lactoglobulin and β-Casein by Trypsin with Consideration of Secondary Masking of Intermediate Polypeptides

Vorob’ev

2022

IJMS

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The opening of protein substrates during degradation by proteases and the corresponding exposure of their internal peptide bonds for a successful enzymatic attack, the so-called demasking effect, was studied for β-lactoglobulin (β-LG) and β-casein (β-CN) hydrolyzed by trypsin. Demasking was estimated by monitoring the redshift in intrinsic tryptophan fluorescence, characterizing the accessibility of polypeptide chains to aqueous medium. The secondary masking of intermediate polypeptides, giving an inverse effect to demasking, caused a restriction of the substrate opening. This led to the limitations in the red shift of fluorescence and the degree of hydrolysis with a long time of hydrolysis of β-LG and β-CN at a constant substrate concentration and reduced trypsin concentrations. The proposed proteolysis model included demasking of initially masked bonds in the protein globule or micelle, secondary masking of intermediate polypeptides, and their subsequent slow demasking. The hydrolysis of peptide bonds was modeled taking into account different hydrolysis rate constants for different peptide bonds. It was demonstrated that demasking competes with secondary masking, which is less noticeable at high trypsin concentrations. Modeling of proteolysis taking into account two demasking processes and secondary masking made it possible to simulate kinetic curves consistent with the experimental data.

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Section: Introductionmentioning

confidence: 99%

Modeling of Proteolysis of β-Lactoglobulin and β-Casein by Trypsin with Consideration of Secondary Masking of Intermediate Polypeptides

Vorob’ev

2022

IJMS

View full text Add to dashboard Cite

show abstract

Tryptic hydrolysis of β-lactoglobulin: A generic approach to describe the hydrolysis kinetic and release of peptides

Cited by 6 publications

References 30 publications

Modeling of Proteolysis of β-Lactoglobulin and β-Casein by Trypsin with Consideration of Secondary Masking of Intermediate Polypeptides

Modeling of Proteolysis of β-Lactoglobulin and β-Casein by Trypsin with Consideration of Secondary Masking of Intermediate Polypeptides

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