Milk Protein Hydrolysis by Actinidin—Kinetic and Thermodynamic Characterisation and Comparison to Bromelain and Papain

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Kaur, Surjit ORCID: 0000-0001-5855-766X, Vasiljevic, Todor ORCID: 0000-0003-1395-7349 and Huppertz, Thom ORCID: 0000-0003-4142-9359 (2023) Milk Protein Hydrolysis by Actinidin—Kinetic and Thermodynamic Characterisation and Comparison to Bromelain and Papain. Foods, 12 (23). ISSN 2304-8158


Plant proteases, including actinidin, papain and bromelain, have been widely used in the food industry but with limited application in dairy systems. This research aimed to establish and compare operational parameters (kinetics, temperature, enzyme type, time and thermodynamics) relevant to the applications of these enzymes in the hydrolysis of whey protein isolates (WPI), whey protein concentrates (WPC) or milk protein concentrates (MPC). The degree of hydrolysis (DH) increased with the rise in temperature, and the maximum DH was achieved at 60 °C for all three dairy systems. The addition of papain resulted in a greater %DH of whey proteins in comparison to bromelain. The cleavage of proteins was clearly time-dependent (p < 0.05), while the pH did not change significantly (p > 0.05) during this time. PAGE analysis revealed that all three enzymes mainly acted on α-lactalbumin and αs-casein in WPI and MPC, respectively. Kinetic parameters from the Lineweaver–Burk plot at 60 °C using WPC and MPC as a substrate varied widely, establishing that WPC hydrolysis was characterised by a lower KM, higher kcat, kcat/KM and Vmax compared to MPC in the case of all three enzymes. The difference in kcat/KM values amongst all enzymes (actinidin > papain > bromelain) indicated the difference in the strength of substrate binding sites. The thermodynamic parameters of these enzymes with MPC and WPC were also determined at a temperature range of 15–60 °C, and the results indicate the potential application of papain and actinidin in the dairy industry.

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Item type Article
DOI 10.3390/foods12234248
Official URL
Subjects Current > FOR (2020) Classification > 3006 Food sciences
Current > Division/Research > Institute for Sustainable Industries and Liveable Cities
Current > Division/Research > College of Health and Biomedicine
Keywords actinidin; papain; bromelain; proteolysis; milk protein concentrate; whey protein concentrate; whey protein isolate; kinetics; thermodynamic parameters
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