Effect of different protease on the hydrolysis degree and antigenicity of cow milk protein

Xiaojun Chu; Ling Li; Ruosi Fang; Guipu Li; Lina Xiong; Donghong Liu

doi:10.1515/ijfe-2024-0136

Abstract

Cow milk protein (CMP) has high nutritional value and is a common food additive, but its allergenicity has raised concerns. In this study, we investigated whey protein and casein hydrolysis as a potential method of lowering CMP antigenicity. We tested three endo-type proteases (neutral protease, papain, trypsin) and one exo-type protease (flavourzyme). We applied response surface methodology to obtain optimized conditions for each enzyme, decreasing antigenicity by >75 %. Maximum hydrolysis of whey protein and casein was achieved with a combination of proteases (1:1.5 papain to flavourzyme, 1:2 and 1:1.5 neutral to flavourzyme in whey protein and casein, respectively). Antigenicity rates declined by 85–92 % across all treatments. The proportion of small-molecular-weight components in enzymatic hydrolysate rose 87–90 %, indicating low antigenicity. Experiments using protein hydrolysates and in vitro simulated digestion tracts revealed decreasing antigenicity as digestion proceeded. Our findings have important implications for the development of hypoallergenic milk products.

Keywords: allergens; casein; hydrolysis; molecular distribution; whey protein

Corresponding author: Donghong Liu, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China, E-mail: dhliu@zju.edu.cn

Funding source: National Key Research and Development Program of China

Award Identifier / Grant number: 019YFE0103900

Funding source: Key Research Project for Agriculture and Social Development of Hangzhou

Award Identifier / Grant number: 202203A11

Research ethics: Not applicable.
Informed consent: Informed consent was obtained from all individuals included in this study.
Author contributions: The authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Use of Large Language Models, AI and Machine Learning Tools: None declared.
Conflict of interest: The authors state no conflict of interest.
Research funding: National Key Research and Development Program of China (019YFE0103900) and the Key Research Project for Agriculture and Social Development of Hangzhou (202203A11).
Data availability: The raw data can be obtained on request from the corresponding author.

Abbreviations and nomenclature

BBD: Box–Behnken design
CMP: Cow milk protein
CMPA: Cow milk protein allergy
DH%: Degree of hydrolysis
MW: Molecular weight
RSM: Response surface methodology

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/ijfe-2024-0136).

Received: 2024-06-20

Accepted: 2024-12-10

Published Online: 2025-02-24

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Supplementary Material