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Application of high-resolution ultrasonic spectroscopy for real-time monitoring of trypsin activity in ß-casein solution.
Melikishvili, Sopio; Dizon, Mark; Hianik, Tibor.
Afiliação
  • Melikishvili S; Department of Nuclear Physics and Biophysics, Comenius University, Mlynska dolina F1, 842 48 Bratislava, Slovakia.
  • Dizon M; School of Chemistry and Chemical Biology, University College of Dublin, Belfield, Dublin 4, Ireland.
  • Hianik T; Department of Nuclear Physics and Biophysics, Comenius University, Mlynska dolina F1, 842 48 Bratislava, Slovakia. Electronic address: tibor.hianik@fmph.uniba.sk.
Food Chem ; 337: 127759, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32777568
ABSTRACT
High-resolution ultrasonic spectroscopy (HR-US) was applied for real-time monitoring of ß-casein hydrolysis by trypsin at various conditions for the first time. The technique is based on the precision measurement of hydration changes proportional to the number of peptide bond hydrolyzed. As HR-US exhibits ultrasonic transparency for most solution, the analysis did not require optical transparency like for 2,4,6-trinitrobenzenesulfonic acid (TNBS) assay. Appropriate enzymatic models were fitted with degree of hydrolysis (dh) profiles to provide kinetic and mechanistic description of proteolysis in terms of initial hydrolysis rate, r0, and rate constant of hydrolysis, kh, and enzyme inactivation, kd. Maximal r0 and dh were obtained at 45 °C and pH 8. The exponential dependence of kinetic parameters allowed determination of the activation (EA = 50.3 ± 7 kJ/mol) and deactivation (ED = 62.23 ± 3 kJ/mol) energies of hydrolysis. The ultrasonic assay provided rapid detection of trypsin activity even at sub-nanomolar concentration.
Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Idioma: Inglês Revista: Food Chem Ano de publicação: 2021 Tipo de documento: Artigo País de afiliação: Eslováquia

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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: MEDLINE Idioma: Inglês Revista: Food Chem Ano de publicação: 2021 Tipo de documento: Artigo País de afiliação: Eslováquia