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Abstract
Background: Different methods for the extraction of trypsin inhibitors in beans (Phaseolus spp.) were investigated. Two randomised complete laboratory experiments were performed, one on the seeds and one on the pods. In the first, the seeds of common bean variety KIS Marcelijan, breeding line Ref_316 × 498 and runner bean variety Bonela were examined. In the second, the fresh pods of five common beans (three breeding lines, two varieties) were analysed. Four extraction methods were used, including ultrasonic-assisted extraction (UAE) for 15 and 30 min and shaking-assisted extraction for 60 and 180 min.
Results: The results showed a significant increase in trypsin inhibitor activity-related traits in UAE compared to shaking extraction, with the 15 min ultrasonic process showing better efficacy than the one with 30 min duration. In the seed experiment, the breeding line Ref_316 × 498 showed the highest Trypsin Units Inhibited (TUI) and TUI/mg sample after a 15 min UAE. In the pod experiment, the breeding line 228_4aa_ca also showed the highest TUI and TUI/mg sample after a 15 min extraction with UAE. These results underline the potential of UAE to maximise trypsin inhibitor content. In addition, remarkable correlations between TUI, TUI/mg sample and the percentage of trypsin inhibition (%TIn) were observed in both experiments.
Conclusions: These results provide valuable insights into the relationship between bean genetic resources, extraction methods and trypsin inhibitor content in bean pods and seeds and serve as a basis for refining extraction protocols. The study encourages further research on the practical implications of investigated protocols for breeding programs and agricultural practices.
References
Raatz B, Mukankusi C, Lobaton JD, et al. Analyses of African common bean (Phaseolus vulgaris L.) germplasm using a SNP fingerprinting platform: Diversity, quality control and molecular breeding. Genet Resour Crop Evol 2019;66(3):707–722. https://doi.org/10.1007/s10722-019-00746-0 PMid: 30956400
Schwember AR, Carrasco B, Gepts P. Unraveling agronomic and genetic aspects of runner bean (Phaseolus coccineus L.). Field Crops Res 2017;206:86–94. https://doi.org/10.1016/j.fcr.2017.02.020
Wu X, Tan M, Zhu Y, et al. The influence of high pressure processing and germination on anti-nutrients contents, in vitro amino acid release and mineral digestibility of soybeans. J Food Comp Anal 2023;115:104953. https://doi.org/10.1016/j.jfca.2022.104953
Divekar PA, Rani V, Majumder S, et al. Protease inhibitors: An induced plant defense mechanism against herbivores. J Plant Growth Regul 2023;42(10):6057-6073. https://doi.org/10.1007/s00344-022-10767-2
do Amaral M, Freitas ACO, Santos AS, et al. TcTI, a Kunitz-type trypsin inhibitor from cocoa associated with defense against pathogens. Sci Rep 2022;12(1):698. https://doi.org/10.1038/s41598-021-04700-y PMid: 35027639
Barbole RS, Saikhedkar N, Giri A. Plant peptides as protease inhibitors for therapeutic and agricultural applications. In: Maheshwari VL, Patil RH, editors. Natural products as enzyme inhibitors: An industrial perspective. Singapore: Springer Nature Singapore; 2022, p.25-57. https://doi.org/10.1007/978-981-19-0932-0_2
Arnaiz A, Talavera-Mateo L, Gonzalez-Melendi P, et al. Arabidopsis Kunitz trypsin inhibitors in defense against spider mites. Frontiers Plant Sci 2018;9:986. https://doi.org/10.3389/fpls.2018.00986 PMid: 30042779
Srinivasan A, Giri AP, Harsulkar AM, et al. A Kunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts anti-metabolic effect on podborer (Helicoverpa armigera) larvae. Plant Mol Biol 2005;57(3):359–374. https://doi.org/10.1007/s11103-004-7925-2 PMid: 15830127
Mittal A, Kansal R, Kalia V, et al. A kidney bean trypsin inhibitor with an insecticidal potential against Helicoverpa armigera and Spodoptera litura. Acta Physiologiae Plantarum 2014;36(2):525–539. https://doi.org/10.1007/s11738-013-1433-4
Zhang A, Li T, Yuan L, et al. Digestive characteristics of Hyphantria cunea larvae on different host plants. Insects 2023;14(5):463. https://doi.org/10.3390/insects14050463 PMid: 37233091
Patriota LL, Procópio TF, De Souza MF, et al. A trypsin inhibitor from Tecoma stans leaves inhibits growth and promotes ATP depletion and lipid peroxidation in Candida albicans and Candida krusei. Frontiers Microbiol 2016;7:611. https://doi.org/10.3389/fmicb.2016.00611 PMid: 27199940
Avilés?Gaxiola S, Chuck?Hernández C, Serna Saldívar SO. Inactivation methods of trypsin inhibitor in legumes: A review. J Food Sci 2018;83(1):17–29. https://doi.org/10.1111/1750-3841.13985 PMid: 29210451
Župunski V, Vasi? M, Šuštar-Vozli? J, et al. Uncertainty of trypsin inhibitor activity measurement of legume crops using microtiter plate method. Food Anal Meth 2018;11(4):1034–1040. https://doi.org/10.1007/s12161-017-1076-y
Bosmali I, Giannenas I, Christophoridou S, et al. Microclimate and genotype impact on nutritional and antinutritional quality of locally adapted landraces of common bean (Phaseolus vulgaris L.). Foods 2023;12(6):1119. https://doi.org/10.3390/foods12061119 PMid: 36981046
Kakade ML, Rackis JJ, McGhee JE, Puski G. Determination of trypsin inhibitor activity of soy products: A collaborative analysis of an improved procedure. Am Assoc Cereal Chem 1974;51(3):376–382. Available from: http://www.aaccnet.org/cerealchemistry/abstracts/1974/cc1974a45.asp [cited December 13, 2023].
Smith C, van Megen W, Twaalfhoven L, et al. The determination of trypsin inhibitor levels in foodstuffs. J Sci Food Agric 1980;31(4):341–350. https://doi.org/10.1002/jsfa.2740310403 PMid: 7190200
Liu K. Soybean trypsin inhibitor assay: Further improvement of the standard method approved and reapproved by American Oil Chemists’ Society and American Association of Cereal Chemists International. J Am Oil Chemists’ Society 2019;96(6):635–645. https://doi.org/10.1002/aocs.12205
Vanga SK, Wang J, Raghavan V. Effect of ultrasound and microwave processing on the structure, in-vitro digestibility and trypsin inhibitor activity of soymilk proteins. Food Sci Technol 2020;131:109708. https://doi.org/10.1016/j.lwt.2020.109708
Giuberti G, Tava A, Mennella G, et al. Nutrients’ and antinutrients’ seed content in common bean (Phaseolus vulgaris L.) lines carrying mutations affecting seed composition. Agronomy 2019;9(6):317. https://doi.org/10.3390/agronomy9060317
Chemat F, Rombaut N, Sicaire AG, et al. Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrasonics Sonochemistry 2017;34:540-560. https://doi.org/10.1016/j.ultsonch.2016.06.035 PMid: 27773280
Singla M, Sit N. Application of ultrasound in combination with other technologies in food processing: a review. Ultrasonics Sonochemistry 2021;73:105506. https://doi.org/10.1016/j.ultsonch.2021.105506 PMid: 33714087
Chandra-Hioe MV, Xu H, Arcot J. The efficiency of ultrasonic-assisted extraction of cyanocobalamin is greater than heat extraction. Heliyon 2020;6(1):e03059. https://doi.org/10.1016/j.heliyon.2019.e03059 PMid: 31909249
Wang J, Li X, Xia X, et al. Extraction, purification, and characterization of a trypsin inhibitor from cowpea seeds (Vigna unguiculata). Preparative Biochem Biotechnol 2014;44(1):1–15. https://doi.org/10.1080/10826068.2013.782041 PMid: 24117148
Wati RK, Theppakorn T, Rawdkuen S. Extraction of trypsin inhibitor from three legume seeds of the Royal Project Foundation. Asian J Food Agro-Industry 2009;2(3):245–254.
Liu K. Trypsin inhibitor assay: Expressing, calculating, and standardizing inhibitor activity in absolute amounts of trypsin inhibited or trypsin inhibitors. J Am Oil Chemists’ Soc 2021;98(4):355–373. https://doi.org/10.1002/aocs.12475
Siqueira LA, Almeida LF, Fernandes JPA, et al. Ultrasonic-assisted extraction and automated determination of catalase and lipase activities in bovine and poultry livers using a digital movie-based flow-batch analyzer. Ultrasonics Sonochemistry 2021;79:105774. https://doi.org/10.1016/j.ultsonch.2021.105774 PMid: 34628308
Liu K, Seegers S, Cao W, et al. An international collaborative study on trypsin inhibitor assay for legumes, cereals, and related products. J Am Oil Chemists’ Soc 2021;98(4):375–390. https://doi.org/10.1002/aocs.12459
Gualberto NC, de Oliveira CS, Nogueira JP, et al. Bioactive compounds and antioxidant activities in the agro-industrial residues of acerola (Malpighia emarginata L.), guava (Psidium guajava L.), genipap (Genipa americana L.) and umbu (Spondias tuberosa L.) fruits assisted by ultrasonic or shaker extraction. Food Res Int 2021;147:110538. https://doi.org/10.1016/j.foodres.2021.110538 PMid: 34399515
Dong J, Liu Y, Liang Z, et al. Investigation on ultrasound-assisted extraction of salvianolic acid B from Salvia miltiorrhiza root. Ultrasonics Sonochemistry 2010;17(1):61–65. https://doi.org/10.1016/j.ultsonch.2009.05.006 PMid: 19497776
Blicharski T, Oniszczuk A. Extraction methods for the isolation of isoflavonoids from plant material. Open Chemistry 2017;15(1):34–45. https://doi.org/10.1515/chem-2017-0005
Jha AK, Sit N. Extraction of bioactive compounds from plant materials using combination of various novel methods: A review. Trends Food Sci Technol 2022;119:579–591. https://doi.org/10.1016/j.tifs.2021.11.019
Nagl N, Sinkovi? L, Savi? A, et al. Trypsin inhibitor activity in grass pea seeds (Lathyrus sativus L.). Ratarstvo i povrtarstvo 2023;60(2):32–39. https://doi.org/10.5937/ratpov60-45934
Shivakumar M, Verma K, Talukdar A, et al. Genetic variability and effect of heat treatment on trypsin inhibitor content in soybean [Glycine max (L.) Merrill.]. Legume Res 2015;38(1):60–65. https://doi.org/10.5958/0976-0571.2015.00010.7
Paniwnyk L, Cai H, Albu S, et al. The enhancement and scale up of the extraction of anti-oxidants from Rosmarinus officinalis using ultrasound. Ultrasonics Sonochemistry 2009;16(2):287–292. https://doi.org/10.1016/j.ultsonch.2008.06.007 PMid: 18778964
Shirsath SR, Sonawane SH, Gogate PR. Intensification of extraction of natural products using ultrasonic irradiations: A review of current status. Chem Engin Process: Process Intensification 2012;53:10–23. https://doi.org/10.1016/j.cep.2012.01.003
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