Enzymatic Modification of Rice Glutelin (Oryza sativa L. japonica Group) for the Development of ACE-Inhibitory Bioactive Peptides
DOI:
https://doi.org/10.14738/aivp.1306.19730Keywords:
rice-glutelin protein, degree of hydrolysis, enzymatic hydrolysis, angiotensin I-converting enzyme (ACE)-inhibitory peptides, antioxidant peptidesAbstract
Bioactive peptides derived from plant seeds have been shown to have various biological activities, particularly in nutraceutical and pharmaceutical applications. The fact that glutelin contributes approximately 80% of total protein content indicates that rice has strong potential to generate a wide variety of bioactive peptides through protein hydrolysis, including the ability to inhibit angiotensin I-converting enzyme (ACE), making it a potential candidate for managing hypertension. In this study, rice glutelin protein was hydrolyzed using three main proteases: pepsin, trypsin, alcalase, and a combination of pepsin and trypsin for five hours. Degree of hydrolysis (DH), ABTS•+ radical scavenging and ACE-inhibitory activity of the hydrolysates were measured. The results indicated the highest DH was obtained with pepsin- and alcalase-based hydrolysates with 23%. Additionally, the ABTS•+ radical scavenging activity of the hydrolysate produced by pepsin+trypsin combination (27.64 ± 0.09 µg/mL) the ACE-inhibitory activity of the hydrolysate produced by alcalase was the highest among the other proteases (5.85 ± 0.01 µg/mL).
References
Roy, T., et al., Rice protein-derived bioactive peptides: Production, purification, health promoting benefits and food applications. J. Food Compos. Anal., 2025. 148: p. 108142.
Bidram, M., and M.R. Ganjalikhany, Bioactive peptides from food science to pharmaceutical industries: Their mechanism of action, potential role in cancer treatment and available resources. Heliyon, 2024.
Samtiya, M., et al., Production, Purification, and Potential Health Applications of Edible Seeds’ Bioactive Peptides: A Concise Review. Foods, 2021. 10(11): p. 2696.
Susanti, E.F.A., E. Susilowati, T.A Siswoyo, Effect of germination period on the antioxidant activities and angiotensin-I converting enzyme inhibitory of Indonesian black rice. Food Res., 2022. 6(4): p. 59–67.
Xinkang L, et al., Rice Storage Proteins: Focus on Composition, Distribution, Genetic Improvement and Effects on Rice Quality. Rice Sci, 2023. 30(3): p. 207–221.
Gan L, et al. Unique Glutelin Expression Patterns and Seed Endosperm Structure Facilitate Glutelin Accumulation in Polyploid Rice Seed. Rice, 2021. 14(1): p. 021.
Zhao, M., et al., Enhancing the solubility and foam ability of rice glutelin by heat treatment at pH12 : Insight into protein structure. Food Hydrocoll., 2020. 103: p. 105626.
Khammuang, S., et al., Antioxidant potential of hydrolyzed proteins from Thai rice varieties and docking studies of novel peptides with free radicals. AIMS Agric. Food, 2025. 10(2): p. 371–389.
Liu, B., et al., Rice protein: Recent advances in composition and protential physiological functions. Food Sci. Hum. Wellness, 2025.
Yan, F., et al., Preparation process optimization and evaluation of bioactive peptides from Carya cathayensis Sarg meal. Curr. Res. Food Sci., 2023. 6: p. 100408.
Vogelsang-O’Dwyer, M., et al., Enzymatic Hydrolysis of Pulse Proteins as a Tool to Improve Techno-Functional Properties. Foods, 2022. 11(9): p. 1307.
Lu, X., et al., Dual-enzyme hydrolysis for preparation of ACE-inhibitory peptides from sesame seed protein: Optimization, separation, and identification. J. Food Biochem., 2021. 45(4): p. 1–18.
Shu, G., et al., Effect of different proteases on the degree of hydrolysis and angiotensin I-converting enzyme-inhibitory activity in goat and cow milk. Biomolecules, 2018. 8(4).
Rudolph, S., et al., Identification and quantification of ACE-inhibiting peptides in enzymatic hydrolysates of plant proteins. Food Chem., 2017. 224: p. 19–25.
Zhu, R., et al., Exploration and molecular mechanism of novel ACE inhibitory peptides from goat milk protein: A combined in silico and in vitro study. Int. Dairy J., 2025. 167: p. 106265.
Ju, Z.Y., N.S. Hettiarachchy, N. Rath, Extraction, denaturation and hydrophobic Properties of Rice Flour Proteins. J. Food Sci., 2001. 66(2): p. 229–232.
Vanvi, A., and A. Tsopmo, Pepsin digested oat bran proteins: separation, antioxidant activity, an identification of new peptides. J. Chem., 2016, 2016(1): p. 1–8.
Awuor, O.L., et al., Optimization of alcalase hydrolysis conditions for production of dagaa (Rastrineobola argenta) protein hydrolysate with antioxidative properties. Ind. Chem., 2017. 3(1): p. 1–6.
Adler-Nissen, J., Determination of the degree of hydrolysis of food protein hydrolysate by trinitrobenzensulfonic acid. J. Agric. Food Chem., 1978. 27(06): p. 1256–1262.
Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 1970. 227(5259): p. 680–685.
Karami, Z., et al., Response Surface Methodology to Optimize Hydrolysis Parameters in Production of Antioxidant Peptides from Wheat Germ Protein by Alcalase Digestion and Identification of Antioxidant Peptides by LC-MS/MS. J. Agr. Sci. Tech., 2019. 21(4): p. 829–844.
Muguruma, M., et al., Identification of pro-drug type ACE inhibitory peptide sourced from porcine myosin B: Evaluation of its antihypertensive effects in vivo. Food Chem., 2008. 114(2): p. 516–522.
da Silva Crozatti, T.T., et al., Obtaining of bioactive di‐ and tripeptides from enzymatic hydrolysis of soybean meal and its protein isolate using Alcalase® and Neutrase®. Int. J. Food Sci. Technol., 2023. 58(3): p. 1586–1596. URL: http://dx.doi.org/10.14738/aivp.1306.19730 263
Rivas-Vela, C.I., et al., Protein hydrolysis by subcritical water: A new perspective on obtaining bioactive peptides. Molecules, 2021. 26(21): p. 1–15.
Pantoa, T., et al., Characterization and bioactivities of young rice protein hydrolysates. J. Cereal Sci., 2020. 95: p. 103049.
Sun, S., et al., Preparation and identification of ACE inhibitory peptides from the marine macroalga Ulva intestinalis. Mar. Drugs, 2019. 17(3): p. 1–17.
Scarabattoli, L., et al., ACE-inhibitory activity and antioxidant properties of a low MW rice bran protein hydrolysate. Lwt., 2025. 217: p. 117381.
Zhang, M., et al., Debittering effect of partially purified proteases from soybean seedlings on soybean protein isolate hydrolysate produced by alcalase. Food Chem., 2021. 362: p. 130190.
Xia, N., et al., Characterization and In Vitro digestibility of rice protein prepared by enzyme-assisted microfluidization: Comparison to alkaline extraction. J. Cereal Sci., 2012. 56(2): p. 482–489.
Yamagata, H., et al., Biosynthesis of Storage proteins in Developing Rice Seeds. Plant Physiol., 1982. 70: p. 1094–1100.
Hwang, J., et al., Comparative Exploration of Antioxidant Properties of Alcalase- and Trypsin-Hydrolyzed Porcine By-Products and Their Classification for Industrial Use. Appl. Sci., 2024. 15(1): p. 47.
Nyo, M.K., and T. Nguyen, Value-Addition of Defatted Peanut Cake by Proteolysis: Effects of Proteases and Degree of Hydrolysis on Functional Properties and Antioxidant Capacity of Peptides. Waste and Biomass Valorization, 2019. 10(5): p. 1251–1259.
Durak, M.Z., and N.A Turan, Antihypertensive Peptides in Dairy Products. AJBSR, 2020. 7(2): p. 191–195.
Supriyadi, A., et al., Revealing antioxidant and antidiabetic potency of melinjo (Gnetum gnemon) seed protein hydrolysate at different stages of seed maturation. Curr. Res. Nutr. Food Sci., 2019. 7(2): p. 479–487.
Siswoyo, T.A., E. Noviyanti, and A. Isnainun, Changes in the antioxidant activities and angiotensin-I converting enzyme inhibitory protein during the germination of green coffee beans. Coffee Sci., 2025. 20: p. e202332
Heo, S.Y., et al., Purification and Molecular Docking Study on the Angiotensin I-Converting Enzyme (ACE)-Inhibitory Peptide Isolated from Hydrolysates of the Deep-Sea Mussel Gigantidas vrijenhoeki. Mar. Drugs, 2023. 21(8).
Jayaprakash, G., et al., A Narrative Review on Rice Proteins: Current Scenario and Food Industrial Application. Polymers (Basel), 2022. 14(15): p. 1–20.
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