Exploring genotypic diversity and processing effects on protein quality for nutritional and functional enhancement in pigeon pea (Cajanus cajan L.)
Main Article Content
Abstract
Pigeon pea protein isolate (PpPI) offers a sustainable and high-quality plant-based protein alternative. This study investigated the in-vitro protein digestibility of thirty pigeon pea genotypes, identifying two contrasting lines—Pusa Arhar 2018-4 (low digestibility) and ICP 1452 (high digestibility). The cytotoxicity, amino acid bioavailability, and gene expression modulation induced by PpPI hydrolysates, along with the impact of thermal processing, were evaluated using Caco-2 cells. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay confirmed that PpPI hydrolysates were non-cytotoxic in both control and autoclaved samples. The amino acid bioavailability assay revealed that neutral and polar amino acids (glycine, alanine, serine, proline, and leucine) had higher transport efficiency (17.43–27.90%) than charged amino acids (9.08–18.76%). Autoclaving significantly improved amino acid transport, with bioavailability ranging from 3.18–21.93% in Pusa Arhar 2018-4 and 4.38–34.84% in ICP 1452. Gene expression analysis using Caco2 cell line showed upregulation of the peptide transporter gene PepT1 (1.04–1.87-fold in Pusa Arhar 2018-4; 1.07–1.89-fold in ICP 1452), with significantly higher expression in autoclaved samples (p <0.05). SREBP2, a key cholesterol metabolism regulator, was downregulated in both genotypes, though not significantly affected by thermal processing. These findings highlight the potential of autoclaved PpPI hydrolysates to improve amino acid bioavailability and intestinal gene expression, providing key insights for selecting pigeon pea genotypes with superior protein quality to guide breeding for enhanced nutritional functionality.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Adibi, S. A. (1997). The oligopeptide transporter (Pept-1) in human intestine: biology and function. Gastroenterology, 113(1), 332-340.
AOAC (2006). Official methods of analysis (18thed.). Washington, DC: The Association of Official Analytical Chemists.
Bakke, S., Jordal, A.-E. O., Gómez-Requeni, P., Verri, T., Kousoulaki, K., Aksnes, A., & Rønnestad, I. (2010). Dietary protein hydrolysates and free amino acids affect the spatial expression of peptide transporter PepT1 in the digestive tract of Atlantic cod (Gadus morhua). Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology, 156(1), 48–55. https://doi.org/10.1016/j.cbpb.2010.02.002
Dutta, M., Dineshkumar, R., Nagesh, C. R., Lakshmi, Y. D., Lekhak, B., Bansal, N., et al. (2024). Exploring protein structural adaptations and polyphenol interactions: Influences on digestibility in pigeon pea dal and whole grains under heat and germination conditions. Food Chemistry, 460, 140561.
Erickson, R. H., Gum, J. R., Lindstrom, M. M., McKean, D., & Kim, Y. S. (1995). Regional expression and dietary regulation of rat small intestinal peptide and amino acid transporter mRNAs. Biochemical and biophysical research communications, 216(1), 249-257.
Feng, M., & Betti, M. (2017). Transepithelial transport efficiency of bovine collagen hydrolysates in a human Caco-2 cell line model. Food Chemistry, 224, 242–250. https://doi.org/10.1016/j.foodchem.2016.12.044.
Görgüç, A., Gençdağ, E., & Yılmaz, F. M. (2020). Bioactive peptides derived from plant origin by-products: Biological activities and techno-functional utilizations in food developments – A review. Food Research International, 136, 109504. https://doi.org/10.1016/j.foodres.2020.109504
Goulart, A. J., Bassan, J. C., Barbosa, O. A., Marques, D. P., Silveira, C. B., Santos, A. F., ... & Monti, R. (2014). Transport of amino acids from milk whey by Caco-2 cell monolayer after hydrolytic action of gastrointestinal enzymes. Food Research International, 63, 62-70.
Hadnađev, M. S., Hadnađev, T. R. D., Pojić, M. M., Šarić, B. M., Mišan, A. Č., Jovanov, P. T., & Sakač, M. B. (2017). Progress in vegetable proteins isolation techniques: A review. Food and Feed research, 44(1), 11-22.
Haji, A., Teka, T. A., Yirga Bereka, T., Negasa Andersa, K., Desalegn Nekera, K., Geleta Abdi, G., Lema Abelti, A., & Makiso Urugo, M. (2024). Nutritional composition, bioactive compounds, food applications, and health benefits of pigeon pea (Cajanus cajan L. Millsp.): A Review. Legume Science, 6(2). https://doi.org/10.1002/leg3.233
Kitts, D., & Weiler, K. (2003). Bioactive proteins and peptides from food sources. applications of bioprocesses used in isolation and recovery. Current Pharmaceutical Design, 9(16), 1309–1323. https://doi.org/10.2174/1381612033454883
Kumar, V., Muthu Kumar, S. P., & Tiku, P. K. (2021). Hypocholesterolemic effect of potent peptide and bioactive fraction from pigeon pea by-products in wistar rats. International Journal of Peptide Research & Therapeutics, 27(4), 2403–2415. https://doi.org/10.1007/s10989-021-10261-5
Kouadio, J. H., Mobio, T. A., Baudrimont, I., Moukha, S., Dano, S. D., & Creppy, E. E. (2005). Comparative study of cytotoxicity and oxidative stress induced by deoxynivalenol, zearalenone or fumonisin B1 in human intestinal cell line Caco-2. Toxicology, 213(1-2), 56–65. doi:10.1016/j.tox.2005.05.010
Liu, X., Shi, J., Yi, J., Zhang, X., Ma, Q., & Cai, S. (2021). The effect of in vitro simulated gastrointestinal digestion on phenolic bioaccessibility and bioactivities of Prinsepia utilis Royle fruits. LWT- Food Science and Technology, 138, 110782. https://doi.org/10.1016/j.lwt.2020.110782
Madison, B. B. (2016). Srebp2: A master regulator of sterol and fatty acid synthesis. Journal of Lipid Research, 57(3), 333–335. https://doi.org/10.1194/jlr.C066712
Nakhle, J., ¨ Ozkan, T., Lnˇ eniˇ ckov´ a, K., Briolotti, P., & Vignais, M.-L. (2020). Methods for simultaneous and quantitative isolation of mitochondrial DNA, nuclear DNA and RNA from mammalian cells, Bio. Techniques, 69(6), 436–442. https://doi.org/ 10.2144/btn-2020-0114
Papalamprou, E. M., Doxastakis, G. I., & Kiosseoglou, V. (2010a). Chickpea protein isolates obtained by wet extraction as emulsifying agents. Journal of the Science of Food & Agriculture, 90(2), 304–313. https://doi.org/10.1002/jsfa.3816
Samtiya, M., Aluko, R. E., & Dhewa, T. (2020). Plant food anti-nutritional factors and their reduction strategies: an overview. Food Production, Processing & Nutrition, 2(1), 6. https://doi.org/10.1186/s43014-020-0020-5
Sen Roy, S., & Seshagiri, P. B. (2013). Expression and function of cyclooxygenase-2 is necessary for hamster blastocyst hatching. Molecular Human Reproduction, 19(12), 838–851. https://doi.org/10.1093/molehr/gat063
Trigo, J. P., Engström, N., Steinhagen, S., Juul, L., Harrysson, H., Toth, G. B., Pavia, H., Scheers, N., & Undeland, I. (2021). In vitro digestibility and Caco-2 cell bioavailability of sea lettuce (Ulva fenestrata) proteins extracted using pH-shift processing. Food Chemistry, 356, 129683. https://doi.org/10.1016/j.foodchem.2021.129683
Wang, B., & Li, B. (2018). Charge and hydrophobicity of casein peptides influence transepithelial transport and bioavailability. Food Chemistry, 245, 646–652. https://doi.org/10.1016/j.foodchem.2017.09.032
Wang, B., & Li, B. (2017). Effect of molecular weight on the transepithelial transport and peptidase degradation of casein-derived peptides by using Caco-2 cell model, 218, 1–8. https://doi.org/10.1016/j.foodchem.2016.08.106.
Zhou, Y., Wang, T., She, X., Wei, Y., Zhou, X., & Xiao, Y. (2022). Pretreatment of buckwheat globulin by ultra-high pressure: Effects on enzymatic hydrolysis and final hydrolysate lipid metabolism regulation capacities. Food Chemistry, 379, 132102. https://doi.org/10.1016/j.foodchem.2022.132102