Production of whey protein ingredients with improved bioactivity based on single and sequential enzymatic hydrolysis reactions

Abstract

•	•WPC-80 hydrolysates (WPHs) were obtained using different proteases.
•	•Bioactivity and cytoprotection of WPHs were assessed.
•	•Calcium-chelating capacity of the hydrolysates was studied by a RSM approach.
•	•WPHs presented interesting bioactive properties which could be used as food ingredients.

References

[1.] RochaJM, GuerraA. On the valorization of lactose and its derivatives from cheese whey as a dairy industry by–product: an overview. Eur Food Res Technol, 2020, 246: 2161-74

[2.] BrandelliA, DaroitDJ, CorrêaAPF. Whey as a source of peptides with remarkable biological activities. Food Res Int, 2015, 73: 149-61

[3.] PatelS. Functional food relevance of whey protein: a review of recent findings and scopes ahead. J Funct Food, 2015, 19: 308-19

[4.] MezaBE, De Piante VicínDA, MarinoF, SihufeGA, PeraltaJM, ZorrillaSE. Characterisation of soluble aggregates from commercial whey protein concentrate suspensions: Effect of protein concentration, pH, and heat treatment conditions. Int J Dairy Technol, 2020, 73(2): 429-36

[5.] DulliusA, GoettertMI, de SouzaCFV. Whey protein hydrolysates as a source of bioactive peptides for functional foods–biotechnological facilitation of industrial scale-up. J Funct Food, 2018, 42: 58-74

[6.] ZhaoL, CaiX, HuangS, WangS, HuangY, HongJ, RaoP. Isolation and identification of a whey protein-sourced calcium-binding tripeptide tyr-asp-thr. Int Dairy J, 2015, 40: 16-23

[7.] JustusA, PereiraDG, IdaEI, KurozawaLE. Combined uses of an endo-and exopeptidase in okara improve the hydrolysates via formation of aglycone isoflavones and antioxidant capacity. LWT Food Sci Technol, 2019, 115: 108467

[8.] XuY, GalanopoulosM, SismourE, RenS, MershaZ, LynchP, AlmutaimiA. Effect of enzymatic hydrolysis using endo-and exo-proteases on secondary structure, functional, and antioxidant properties of chickpea protein hydrolysates. J Food Meas Charact, 2020, 14(1): 343-52

[9.] KimatuB, ZhaoL, BiaoY, MaG, YangW, PeiF, HuQ. Antioxidant potential of edible mushroom (Agaricus Bisporus) protein hydrolysates and their ultrafiltration fractions. Food Chem, 2017, 230: 58-67

[10.] EberhardtA, LópezEC, CerutiRJ, MarinoF, MammarellaEJ, ManzoRM, SihufeGA. Influence of the degree of hydrolysis on the bioactive properties of whey protein hydrolysates using Alcalase^®. Int J Dairy Technol, 2019, 72(4): 573-84

[11.] CaiX, ZhaoL, WangS, RaoP. Fabrication and characterization of the nano-composite of whey protein hydrolysate chelated with calcium. Food Function, 2015, 6(3): 816-23

[12.] HouH, WangS, ZhuX, LiQ, FanY, ChengD, LiB. A novel calcium-binding peptide from Antarctic krill protein hydrolysates and identification of binding sites of calcium-peptide complex. Food Chem, 2018, 243: 389-95

[13.] HayakariM, KondoY, IzumiH. A rapid and simple spectrophotometric assay of angiotensin-converting enzyme. Anal Biochem, 1978, 84(2): 361-9

[14.] AshaoluTJ, LeTD, SuttikhanaI. Stability and bioactivity of peptides in food matrices based on processing conditions. Food Res Int, 2023, 168: 112786

[15.] ReR, PellegriniN, ProteggenteA, PannalaA, YangM, Rice-EvansC. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Bio Med, 1999, 26(9–10): 1231-7

[16.] BallatoreMB, BettiolMR, Vanden BraberNL, AminahuelCA, RossiYE, PetroselliG, Erra-BalsellsR, CavaglieriLR, MontenegroMA. Antioxidant and cytoprotective effect of peptides produced by hydrolysis of whey protein concentrate with trypsin. Food Chem, 2020, 319: 126472

[17.] NchienziaHA, MorawickiRO, GadangVP. Enzymatic hydrolysis of poultry meal with endo-and exopeptidases. Poult Sci, 2010, 89(10): 2273-80

[18.] EberhardtA, LópezEC, MarinoF, MammarellaEJ, ManzoRM, SihufeGA. Whey protein hydrolysis with microbial proteases: determination of kinetic parameters and bioactive properties for different reaction conditions. Int J Dairy Technol, 2021, 74(3): 489-504

[19.] AmbigaipalanP, Al-KhalifaAS, ShahidiF. Antioxidant and angiotensin I converting enzyme (ACE) inhibitory activities of date seed protein hydrolysates prepared using Alcalase, Flavourzyme and Thermolysin. J Funct Food, 2015, 18: 1125-37

[20.] ChirinosR, OchoaK, Aguilar-GalvezA, CarpentierS, PedreschiR, CamposD. Obtaining of peptides with in vitro antioxidant and angiotensin I converting enzyme inhibitory activities from cañihua protein (Chenopodium pallidicaule Aellen). J Cereal Sci, 2018, 83: 139-46

[21.] BuT, ZhouM, ZhengJ, YangP, SongH, LiS, WuJ. Preparation and characterization of a low-phenylalanine whey hydrolysate using two-step enzymatic hydrolysis and macroporous resin adsorption. LWT, 2020, 132: 109753

[22.] SunN, WuH, DuM, TangY, LiuH, FuY, ZhuB. Food protein-derived calcium chelating peptides: a review. Trends Food Sci Technol, 2016, 58: 140-8

[23.] WangL, MaoX, ChengX, XiongX, RenF. Effect of enzyme type and hydrolysis conditions on the in vitro angiotensin I-converting enzyme inhibitory activity and ash content of hydrolysed whey protein isolate. Int J Food Sci Technol, 2010, 45(4): 807-12

[24.] XueL, YinR, HowellK, ZhangP. Activity and bioavailability of food protein-derived angiotensin‐I‐converting enzyme–inhibitory peptides. Comp Rev Food Sci F, 2021, 20(2): 1150-87

[25.] WuJ, DingX. Characterization of inhibition and stability of soy-protein-derived angiotensin I-converting enzyme inhibitory peptides. Food Res Int, 2002, 35(4): 367-75

[26.] EscuderoE, MoraL, ToldráF. Stability of ACE inhibitory ham peptides against heat treatment and in vitro digestion. Food Chem, 2014, 161: 305-11

[27.] DryákováA, PihlantoA, MarnilaP, ČurdaL, KorhonenHJ. Antioxidant properties of whey protein hydrolysates as measured by three methods. Eur Food Res Technol, 2010, 230: 865-74

[28.] Mann B, Kumari A, Kumar R, Sharma R, Prajapati K, Mahboob S, Athira S. (2015). Antioxidant activity of whey protein hydrolysates in milk beverage system. J Food Sci Technol. 2015;52:3235–3241. https://doi.org/10.1007/s13197-014-1361-3

[29.] VaštagŽ, PopovićL, PopovićS, KrimerV, PeričinD. Production of enzymatic hydrolysates with antioxidant and angiotensin-I converting enzyme inhibitory activity from pumpkin oil cake protein isolate. Food Chem, 2011, 124(4): 1316-21

[30.] AlahyaribeikS, SharifiSD, TabandehF, HonarbakhshS, GhazanfariS. Stability and cytotoxicity of DPPH inhibitory peptides derived from biodegradation of chicken feather. Prot Expr Purif, 2021, 177: 105748

[31.] CorrochanoAR, ArranzE, De NoniI, StuknytėM, FerrarettoA, KellyPM, BuckinV, GiblinL. Intestinal health benefits of bovine whey proteins after simulated gastrointestinal digestion. J Funct Foods, 2018, 49: 526-35

[32.] O’KeeffeMB, FitzGeraldRJ. Antioxidant effects of enzymatic hydrolysates of whey protein concentrate on cultured human endothelial cells. Int Dairy J, 2014, 36: 128e135

Funding

Universidad Nacional del Litoral(project CAI+D: 506 201901 00043 LI); Consejo Nacional de Investigaciones Científicas y Técnicas(project CONICET: 11220200100592CO)