Abstract
Background: Food proteins have benefits for human health, which justifies their production and use. In this context, the use of seeds and byproducts that would be otherwise discarded is highlighted in the present work, to produce protein extracts and hydrolyzed proteins, generating opportunities to reduce environmental impacts.
Objective: This work aimed to use different extraction methods to obtain protein extracts from seeds (corn, sorghum, and sunflower) and sunflower byproduct to determine their antioxidant activity, and apply different proteolytic enzymes in the hydrolysis of sunflower byproduct.
Methods: The seeds of corn, sorghum, and sunflower, and sunflower byproduct were ground to produce flour and the protein extracts were prepared using five different methods. The bioactivity of fractions was analyzed by different methods (ABTS, DPPH, and FRAP) to evaluate the antioxidant activity.
Results: The most effective methods, which resulted in higher protein extraction and antioxidant activity, were those in which NH4HCO3 (5 mM, pH 8.0) and H2O/C2H6O (2:3) were used. The highest protein contents were 797.9, 303.8, and 11296.5 μg/g, and the highest antioxidant activity was 34417.5, 9732.6, and 47473.1 μg TE/g from Soxhlet and Bligh and Dyer defatted extractions for sunflower seed, and sunflower byproduct, respectively. Regarding enzymatic hydrolysis, sunflower byproduct was the substrate that presented the highest degree of hydrolysis (11.06%) when Neutrase ® enzyme was used. Enzymatic hydrolysis increased antioxidant activity in the hydrolyzed proteins, approximately by 20.0%, using Neutrase® and 22.3% using Flavourzyme® treatments.
Conclusion: The protein extracts and the hydrolyzed proteins exhibited high antioxidant activity, demonstrating great potential for use as natural antioxidants in food systems.
Keywords: Extraction methods, protein extraction, hydrolysis, proteolytic enzymes, byproducts, natural antioxidants.
Graphical Abstract
[1]
Barlow S, Schlatter J. Risk assessment of carcinogens in food. Toxicol Appl Pharmacol 2010; 243(2): 180-90.
[http://dx.doi.org/10.1016/j.taap.2009.11.004] [PMID: 19909764]
[http://dx.doi.org/10.1016/j.taap.2009.11.004] [PMID: 19909764]
[2]
Suja KP, Abraham JT, Thamizh SN, Jayalekshmy A, Arumughan C. Antioxidant efficacy of sesame cake extract in vegetable oil protection. Food Chem 2004; 84(3): 393-400.
[http://dx.doi.org/10.1016/S0308-8146(03)00248-6]
[http://dx.doi.org/10.1016/S0308-8146(03)00248-6]
[3]
Oroian M, Escriche I. Antioxidants: characterization, natural sources, extraction and analysis. Food Res Int 2015; 74: 10-36.
[http://dx.doi.org/10.1016/j.foodres.2015.04.018] [PMID: 28411973]
[http://dx.doi.org/10.1016/j.foodres.2015.04.018] [PMID: 28411973]
[4]
Shen S, Chahal B, Majumder K, You SJ, Wu J. Identification of novel antioxidative peptides derived from a thermolytic hydrolysate of ovotransferrin by LC-MS/MS. J Agric Food Chem 2010; 58(13): 7664-72.
[http://dx.doi.org/10.1021/jf101323y] [PMID: 20568771]
[http://dx.doi.org/10.1021/jf101323y] [PMID: 20568771]
[5]
Asimi OA, Sahu NP, Pal AK. Antioxidant activity and antimicrobial property of some Indian spices. Int J Sci Res 2013; 3(3): 1-8.
[6]
Lemes AC, Sala L, Ores JdaC, Braga AR, Egea MB, Fernandes KF. A review of the latest advances in encrypted bioactive peptides from protein-rich waste. Int J Mol Sci 2016; 17(6): 1-24.
[http://dx.doi.org/10.3390/ijms17060950] [PMID: 27322241]
[http://dx.doi.org/10.3390/ijms17060950] [PMID: 27322241]
[7]
de Oliveira Filho JG, Rodrigues JM, Valadares ACF, et al. Bioactive Properties of Protein Hydrolysate of Cottonseed Byproduct: Antioxidant, Antimicrobial, and Angiotensin-Converting Enzyme (ACE) Inhibitory Activities. Waste Biomass Valoriz 2020; 1-10.
[8]
Xue Z, Yu W, Liu Z, Wu M, Kou X, Wang J. Preparation and antioxidative properties of a rapeseed (Brassica napus) protein hydrolysate and three peptide fractions. J Agric Food Chem 2009; 57(12): 5287-93.
[http://dx.doi.org/10.1021/jf900860v] [PMID: 19432452]
[http://dx.doi.org/10.1021/jf900860v] [PMID: 19432452]
[9]
Zhang L, Li J, Zhou K. Chelating and radical scavenging activities of soy protein hydrolysates prepared from microbial proteases and their effect on meat lipid peroxidation. Bioresour Technol 2010; 101(7): 2084-9.
[http://dx.doi.org/10.1016/j.biortech.2009.11.078] [PMID: 20015640]
[http://dx.doi.org/10.1016/j.biortech.2009.11.078] [PMID: 20015640]
[10]
Sarmadi BH, Ismail A. Antioxidative peptides from food proteins: a review. Peptides 2010; 31(10): 1949-56.
[http://dx.doi.org/10.1016/j.peptides.2010.06.020] [PMID: 20600423]
[http://dx.doi.org/10.1016/j.peptides.2010.06.020] [PMID: 20600423]
[11]
Lemes AC, Paula LC, Oliveira Filho JG, Prado DMF, Medronha GA, Egea MB. Bioactive peptides with antihypertensive property obtained from agro-industrial by products-Mini review. Austin J Nutr Metab 2020.
[12]
Mahatmanto T, Poth AG, Mylne JS, Craik DJ. A comparative study of extraction methods reveals preferred solvents for cystine knot peptide isolation from Momordica cochinchinensis seeds. Fitoterapia 2014; 95: 22-33.
[http://dx.doi.org/10.1016/j.fitote.2014.02.016] [PMID: 24613804]
[http://dx.doi.org/10.1016/j.fitote.2014.02.016] [PMID: 24613804]
[13]
Smet K, Raes K, De Block J, Herman L, Dewettinck K, Coudijzer K. A change in antioxidative capacity as a measure of onset to oxidation in pasteurized milk. Int Dairy J 2008; 18(5): 520-30.
[http://dx.doi.org/10.1016/j.idairyj.2007.11.012]
[http://dx.doi.org/10.1016/j.idairyj.2007.11.012]
[15]
Oseguera-Toledo ME, de Mejia EG, Dia VP, Amaya-Llano SL. Common bean (Phaseolus vulgaris L.) hydrolysates inhibit inflammation in LPS-induced macrophages through suppression of NF-κB pathways. Food Chem 2011; 127(3): 1175-85.
[http://dx.doi.org/10.1016/j.foodchem.2011.01.121] [PMID: 25214111]
[http://dx.doi.org/10.1016/j.foodchem.2011.01.121] [PMID: 25214111]
[16]
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193(1): 265-75.
[PMID: 14907713]
[PMID: 14907713]
[17]
Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. Lebensm Wiss Technol 1995; 28(1): 25-30.
[http://dx.doi.org/10.1016/S0023-6438(95)80008-5]
[http://dx.doi.org/10.1016/S0023-6438(95)80008-5]
[18]
He R, Girgih AT, Malomo SA, Ju X, Aluko RE. Antioxidant activities of enzymatic rapeseed protein hydrolysates and the membrane ultrafiltration fractions. J Funct Foods 2013; 5(1): 219-27.
[http://dx.doi.org/10.1016/j.jff.2012.10.008]
[http://dx.doi.org/10.1016/j.jff.2012.10.008]
[19]
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 1999; 26(9-10): 1231-7.
[http://dx.doi.org/10.1016/S0891-5849(98)00315-3] [PMID: 10381194]
[http://dx.doi.org/10.1016/S0891-5849(98)00315-3] [PMID: 10381194]
[20]
Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 1996; 239(1): 70-6.
[http://dx.doi.org/10.1006/abio.1996.0292] [PMID: 8660627]
[http://dx.doi.org/10.1006/abio.1996.0292] [PMID: 8660627]
[21]
Hernández-Ledesma B, Amigo L, Martín-Álvarez PJ, Recio I. Production of antioxidant hydrolyzates from a whey protein concentrate with thermolysin: optimization by response surface methodology. Lebensm Wiss Technol 2011; 44(1): 9-15.
[http://dx.doi.org/10.1016/j.lwt.2010.06.017]
[http://dx.doi.org/10.1016/j.lwt.2010.06.017]
[22]
Zhong F, Liu J, Ma J, Shoemaker CF. Preparation of hypocholesterol peptides from soy protein and their hypocholesterolemic effect in mice. Food Res Int 2007; 40(6): 661-7.
[http://dx.doi.org/10.1016/j.foodres.2006.11.011]
[http://dx.doi.org/10.1016/j.foodres.2006.11.011]
[23]
Branlard G, Bancel E. Protein extraction from cereal seeds. Methods Mol Biol 2007; 355: 15-25.
[PMID: 17093298]
[PMID: 17093298]
[24]
Carrao-Panizzi MC, Mandarino JMG. 1994; Available at: https://www.embrapa.br/soja/busca-de-publicacoes/-/publicacao/445734/girassol-derivados-proteicos
[26]
Kinsella JE, Melachouris N. Functional properties of proteins in foods: A survey. CRC Crit Rev Food Sci Nutr 1976; 7(3): 219-80.
[http://dx.doi.org/10.1080/10408397609527208]
[http://dx.doi.org/10.1080/10408397609527208]
[27]
Hettiarachchy NS, Ziegler GR. Structure-function relationship of food proteins. Boca Raton: CRC Press In: 1994.
[28]
Emmambux MN, Taylor JR. Properties of heat-treated sorghum and maize meal and their prolamin proteins. J Agric Food Chem 2009; 57(3): 1045-50.
[http://dx.doi.org/10.1021/jf802672e] [PMID: 19143536]
[http://dx.doi.org/10.1021/jf802672e] [PMID: 19143536]
[29]
Argos P, Pedersen K, Marks MD, Larkins BA. A structural model for maize zein proteins. J Biol Chem 1982; 257(17): 9984-90.
[PMID: 7107620]
[PMID: 7107620]
[30]
Anderson TJ, Lamsal BP. Review: zein extraction from corn, corn products, and coproducts and modifications for various applications: a review. Cereal Chem 2011; 88(2): 159-73.
[http://dx.doi.org/10.1094/CCHEM-06-10-0091]
[http://dx.doi.org/10.1094/CCHEM-06-10-0091]
[31]
Espinosa-Ramírez J, Serna-Saldívar SO. Functionality and characterization of kafirin-rich protein extracts from different whole and decorticated sorghum genotypes. J Cereal Sci 2016; 70: 57-65.
[http://dx.doi.org/10.1016/j.jcs.2016.05.023]
[http://dx.doi.org/10.1016/j.jcs.2016.05.023]
[32]
Souza RSC, Balbuenac TS, Arruda P. Structure, organization, and expression of the alpha prolamin multigenic family bring new insights into the evolutionary relationships among grasses. Plant Genome Abstract 2015; 8(1): 1-11.
[33]
Elias RJ, Kellerby SS, Decker EA. Antioxidant activity of proteins and peptides. Crit Rev Food Sci Nutr 2008; 48(5): 430-41.
[http://dx.doi.org/10.1080/10408390701425615] [PMID: 18464032]
[http://dx.doi.org/10.1080/10408390701425615] [PMID: 18464032]
[34]
Carrasco-Castilla J, Hernández-Álvarez AJ, Jiménez-Martínez C, et al. Antioxidant and metal chelating activities of Phaseolus vulgaris L. var. Jamapa protein isolates, phaseolin and lectin hydrolysates. Food Chem 2012; 131(4): 1157-64.
[http://dx.doi.org/10.1016/j.foodchem.2011.09.084]
[http://dx.doi.org/10.1016/j.foodchem.2011.09.084]
[35]
MacDonald-Wicks LK, Wood LG, Garg ML. Methodology for the determination of biological antioxidant capacity in vitro: a review. J Sci Food Agric 2006; 86(13): 2046-56.
[http://dx.doi.org/10.1002/jsfa.2603]
[http://dx.doi.org/10.1002/jsfa.2603]
[36]
Lemes AC, Álvares GT, Egea MB, Brandelli A, Kalil SJ. Simultaneous production of proteases and antioxidant compounds from agro-industrial by-products. Bioresour Technol 2016; 222: 210-6.
[http://dx.doi.org/10.1016/j.biortech.2016.10.001] [PMID: 27718403]
[http://dx.doi.org/10.1016/j.biortech.2016.10.001] [PMID: 27718403]
[37]
Zhu YP, Fan JF, Cheng YQ, Li LT. Improvement of the antioxidant activity of Chinese traditional fermented okara (Meitauza) using Bacillus subtilis B2. Food Control 2008; 19(7): 654-61.
[http://dx.doi.org/10.1016/j.foodcont.2007.07.009]
[http://dx.doi.org/10.1016/j.foodcont.2007.07.009]
[38]
Ramakrishnan V, Goveas LC, Suralikerimath N, Jampani C, Halami PM, Narayan B. Extraction and purification of lipase from Enterococcus faecium MTCC5695 by PEG/phosphate aqueous-two phase system (ATPS) and its biochemical characterization. Biocatal Agric Biotechnol 2016; 6: 19-27.
[http://dx.doi.org/10.1016/j.bcab.2016.02.005]
[http://dx.doi.org/10.1016/j.bcab.2016.02.005]
[39]
Ungcharoenwiwat P. H-Kittikun A. Purification and characterization of lipase from Burkholderia sp. EQ3 isolated from wastewater from a canned fish factory and its application for the synthesis of wax esters. J Mol Catal, B Enzym 2015; 115: 96-104.
[http://dx.doi.org/10.1016/j.molcatb.2015.02.005]
[http://dx.doi.org/10.1016/j.molcatb.2015.02.005]
[40]
Rockenbach II. Silva GLd, Rodrigues E, Kuskoski EM, Fett R. Solvent Influence on total polyphenol content, anthocyanins, and antioxidant activity of grape (Vitis vinifera) bagasse extracts from Tannat and Ancelota - different varieties of Vitis vinifera varieties. Food Sci Technol (Campinas) 2008; 28: 238-44.
[http://dx.doi.org/10.1590/S0101-20612008000500036]
[http://dx.doi.org/10.1590/S0101-20612008000500036]
[41]
Oliveira GLS. Determinação da capacidade antioxidante de produtos naturais in vitro pelo método do DPPH”: estudo de revisão. Rev Bras Plantas Med 2015; 17: 36-44.
[http://dx.doi.org/10.1590/1983-084X/12_165]
[http://dx.doi.org/10.1590/1983-084X/12_165]
[42]
You L, Zhao M, Regenstein JM, Ren J. Changes in the antioxidant activity of loach (Misgurnus anguillicaudatus) protein hydrolysates during a simulated gastrointestinal digestion. Food Chem 2010; v.120 no.3(no. 3): 810-6.
[43]
Alves RE, de Brito ES, Pérez-Jiménez J, Saura-Calixto F, Mancini-Filho J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chem 2010; 121(4): 996-1002.
[http://dx.doi.org/10.1016/j.foodchem.2010.01.037]
[http://dx.doi.org/10.1016/j.foodchem.2010.01.037]
[44]
Luna-Vital DA, Mojica L, González de Mejía E, Mendoza S, Loarca-Piña G. Biological potential of protein hydrolysates and peptides from common bean (Phaseolus vulgaris L.): a review. Food Res Int 2015; 76: 39-50.
[http://dx.doi.org/10.1016/j.foodres.2014.11.024]
[http://dx.doi.org/10.1016/j.foodres.2014.11.024]
[45]
Xia Y, Bamdad F, Gänzle M, Chen L. Fractionation and characterization of antioxidant peptides derived from barley glutelin by enzymatic hydrolysis. Food Chem 2012; 134(3): 1509-18.
[http://dx.doi.org/10.1016/j.foodchem.2012.03.063] [PMID: 25005974]
[http://dx.doi.org/10.1016/j.foodchem.2012.03.063] [PMID: 25005974]
[46]
Betancur-Ancona D, Sosa-Espinoza T, Ruiz-Ruiz J, Segura-Campos M, Chel-Guerrero L. Enzymatic hydrolysis of hard-to-cook bean (Phaseolus vulgaris L.) protein concentrates and its effects on biological and functional properties. Int J Food Sci Technol 2014; 49(1): 2-8.
[http://dx.doi.org/10.1111/ijfs.12267]
[http://dx.doi.org/10.1111/ijfs.12267]
[47]
Vieira EF, da Silva DD, Carmo H, Ferreira IM. Protective ability against oxidative stress of brewers’ spent grain protein hydrolysates. Food Chem 2017; 228: 602-9.
[http://dx.doi.org/10.1016/j.foodchem.2017.02.050] [PMID: 28317769]
[http://dx.doi.org/10.1016/j.foodchem.2017.02.050] [PMID: 28317769]
[48]
Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005; 53(6): 1841-56.
[http://dx.doi.org/10.1021/jf030723c] [PMID: 15769103]
[http://dx.doi.org/10.1021/jf030723c] [PMID: 15769103]
Article Metrics
5