Evaluation inhibitory activity of catechins on trypsin by capillary electrophoresis-based immobilized enzyme microreactor with chromogenic substrate

© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim..

In this study, a capillary electrophoresis-based online immobilized enzyme microreactor was developed for evaluating the inhibitory activity of green tea catechins and tea polyphenol extracts on trypsin. The immobilized trypsin activity and other kinetic parameters were evaluated by measuring the peak area of the hydrolyzate of chromogenic substrate S-2765. The results indicated that the activity of the immobilized trypsin remained approximately 90.0% of the initial immobilized enzyme activity after 30 runs. The value of Michaelis-Menten constant (Km ) was (0.47 ± 0.08) mM, and the half-maximal inhibitory concentration (IC50 ) and inhibition constant (Ki ) of benzamidine were measured as 3.34 and 3.00 mM, respectively. Then, the inhibitory activity of four main catechins (epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate) and three tea polyphenol extracts (green tea, white tea, and black tea) on trypsin were investigated. The results showed that four catechins and three tea polyphenol extracts had potential trypsin inhibitory activity. In addition, molecular docking results illustrated that epigallocatechin gallate, epicatechin gallate, epicatechin, and epigallocatechin were all located not only in the catalytic cavity, but also in the substrate-binding pocket of trypsin. These results indicated that the developed method is an effective tool for evaluating inhibitory activity of catechins on trypsin.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:43

Enthalten in:

Journal of separation science - 43(2020), 15 vom: 28. Aug., Seite 3136-3145

Sprache:

Englisch

Beteiligte Personen:

Zhang, Hao [VerfasserIn]
Lu, Min [VerfasserIn]
Jiang, Hui [VerfasserIn]
Wang, Xu [VerfasserIn]
Yang, Feng-Qing [VerfasserIn]

Links:

Volltext

Themen:

60457-00-3
8R1V1STN48
Benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide
Catechin
Catechins
Chromogenic substrates
EC 3.4.21.4
Enzyme Inhibitors
Enzymes, Immobilized
Immobilized enzyme microreactor
Journal Article
Oligopeptides
Plant Extracts
Polyphenols
Tea
Trypsin

Anmerkungen:

Date Completed 23.06.2021

Date Revised 23.06.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1002/jssc.202000132

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM310953162