Separation and characterization of a water soluble protein fraction from Moringa oleifera seeds

Cristóbal Lárez Velásquez

Abstract


A simple and relatively inexpensive method was developed in order to separate and purify the water soluble fraction of proteins present in dry dehulled Moringa seeds directly collected from local trees. Procedure includes: (a) extraction of the oil fraction from M. oleifera seeds using petroleum ether at room temperature; (b) aqueous extraction of defatted seeds; (c) fractionation of the resulting aqueous phase by cooling at 4 oC and separating the precipitated solid by centrifugation; and (d) freezing and vacuum drying of the wet solid. Protein fraction obtained consists of a mixture of molecular species of different sizes, with those around 26.5, 21.0 and < 14.2 KDa present in greater proportion, which shows interesting acid/base properties. Moreover, two important aspects have emerged from analyses performed to this protein sample: (a) a slightly acidic character was observed by potentiometric and conductimetric titrations which could be justified by the presence of tyrosine phenolic groups in the protein chains forming the mixture, as it has been inferred by FTIR studies, and (b) bathochromic displacements observed during UV-visible studies for the signal associated to π → π* transitions of the peptide bonds can be attributed to conformational changes which modify natural α-helix structure of the protein, especially those caused by rupture of hydrogen bonds probably involving phenolic groups of tyrosine.

Keywords


Moringa seeds; Protein conformational changes; Conductimetric/Potentiometric titrations; FTIR characterization; Electrophoretic profiles

Full Text:

PDF

References


Martín, C., Martín, C., García, A., Fernández, T., Hernández, E., Puls, J. (2013). Potenciales aplicaciones de Moringa oleifera. Una revisión crítica. Pastos y Forrajes, 36(2), 137-149. http://scielo.sld.cu/pdf/pyf/v36n2/en_pyf01213.pdf

Ghosh, A., Bhattacharya, R., Pradhan, C., Chaudhuri, K., Mukhopadhyay, A., Bose, C.K. (2016). Antiproliferative effect of Moringa oleifera root extract on ovarian carcinoma: An in vitro study. Annals of Oncology, 27(9), ix94-ix103.

http://oncologypro.esmo.org/Meeting-Resources/ESMO-Asia-2016-Congress/Antiproliferative-effect-of-Moringaoleifera-root-extract-on-ovarian-carcinoma-An-in-vitro-study

Abiodun, B., Adedeji, A., Taiwo, O., Gbenga, A. (2015). Effects of Moringa oleifera root extract on the performance and serum biochemistry of Escherichia coli challenged broiler chicks. J. Agric. Sci., 60(4), 505-513. http://joas.agrif.bg.ac.rs/sites/joas.agrif.bg.ac.rs/files/article/pdf/432-10-759._bolu_et_al._final.pdf

Sivarathnakumar, S., Baskar, G., Kumar, R., Bharathiraja, B. (2016). Bioethanol production by the utilisation of Moringa oleifera stem with sono-assisted acid/alkali hydrolysis approach. Int. J. Environm. Sust. Develop., 15(4), 392-403. https://doi.org/10.1504/IJESD.2016.079481

Singhal, M., Pandey, S., Jijhotiya, A. (2016). Antidiabetic activity of stem extracts of plant Moringa oleifera. Int. Education and Research J., 2(3), 72-73. http://ierj.in/journal/index.php/ierj/article/view/185/167

Lalas, S., Athanasiadis, V., Karageorgou, I., Batra, G., Nanos, G., Makris, D. (2017). Nutritional Characterization of Leaves and Herbal Tea of Moringa oleifera Cultivated in Greece. Journal of Herbs, Spices & Medicinal Plants. http://www.tandfonline.com/doi/abs/10.1080/10496475.2017.1334163

Martínez-González, C., Martínez, L., Martínez-Ortiz, E., González-Trujano, M., Déciga-Campos, M., Ventura-Martínez, R., Díaz-Reval, I. (2017). Moringa oleifera, a species with potential analgesic and anti-inflammatory activities. Biomedicine & Pharmacotherapy, 87, 482–488. https://www.sciencedirect.com/science/article/pii/S0753332216320984

Kalappurayil, T.M. & Joseph, B.P. (2017). A Review of Pharmacognostical Studies on Moringa oleifera Lam. flowers. Pharmacognosy Journal, 9(1), 1-7. http://journalview.org/phcogj.com/article/217

Abioye, V., & Aka, M. (2015). Proximate composition and sensory properties of moringa fortified maize-ogi. Nutr. Food Sci., S12:001. http://agris.fao.org/agris-search/search.do?recordID=US2016B00826

Wuana, R., Sha’Ato, R., Iorhen, S. (2015). Preparation, characterization, and evaluation of Moringa oleifera pod husk adsorbentsfor aqueous phase removal of norfloxacin. Desalination and Water Treatment, 57, 11904-11916. https://doi.org/10.1080/19443994.2015.1046150

Faizi, S., Siddiqui, B., Saleem, R., Aftab, K., Shaheen, F. (1998). Hypotensive constituents from the pods of Moringa oleifera. Planta Medica, 64(3), 225-228. https://doi.org/10.1055/s-2006-957414

Ayerza, R. (2012). Seed and oil yields of Moringa oleifera variety Periyakalum-1 introduced for oil production in four ecosystems of South America. Industrial Crops & Products, 36(1), 70-73. https://doi.org/10.1016/j.indcrop.2011.08.008

Lopes Muniz, G., Veloso Duarte, F., Barbosa de Oliveira, S. (2015). Uso de sementes de Moringa oleifera na remoção da turbidez de água para abastecimento. Rev. Ambient. Agua, 10(2), 254-263. http://www.scielo.br/pdf/ambiagua/v10n2/1980-993X-ambiagua-10-02-00454.pdf

Sengupta, M., Keraita, B., Olsen, A., Boateng, O., Thamsborg, S., Pálsdóttir, G., Dalsgaard, A. (2012). Use of seed extracts to reduce helminth egg numbers and turbidity in irrigation water. Water Research, 46(11), 3646-3656. https://doi.org/10.1016/j.watres.2012.04.011

Tie, T., Li, P., Xu, Z., Zhou, Y., Li, C., Zhang, X. (2015). Removal of Congo red from aqueous solution using Moringa oleifera seed cake as natural coagulant. Desalination and Water Treatment, 54(10), 2817-2824. https://doi.org/10.1080/19443994.2014.905980

Gonçalves-Junior, A., Meneghel, A., Rubio, F., Strey, L., Dragunski, D., Coelho, G. (2013). Applicability of Moringa oleifera Lam. pie as an adsorbent for removal of heavy metals from waters. Revista Brasileira de Engenharia Agrícola e Ambiental, 17(1), 94-99. http://dx.doi.org/10.1590/S1415-43662013000100013

Santos, A., Matos, M., Sousa, A., Costa, C., Nogueira, R., Teixeira, J., Paiva, P., Parpot, P., Coelho, L., Brito, A. (2015). Removal of tetracycline from contaminated water by Moringa oleifera seed preparations. Environm. Technol., 37(6), 744-751. https://doi.org/10.1080/09593330.2015.1080309

Endut, A., Hamid, S., Lananan, F., Khatoon, H. (2016). Moringa oleifera seed derivatives as potential bio-coagulant for microalgae Chlorella Sp. harvesting. Malaysian Journal of Analytical Sciences, 20(2), 401-412. https://www.researchgate.net/profile/Fathurrahman_Lananan/publication/301362302_Moringa_oleifera_seed_derivatives_as_potential_bio-coagulant_for_microalgae_Chlorella_sp_harvesting/links/5715d35408ae1a840265070a.pdf

Bichi, M.H. (2013). A review of the applications of Moringa oleifera seeds extract in water treatment. Civil and Environmental Research, 3(8), 1-10. http://www.academia.edu/download/31667219/A_Review_of_the_Applications_of_Moringa_oleifera_Seeds_Extract_in_Water_Treatment.pdf

Nordmark, B., Przybycien, T., Tilton, R. (2016). Comparative coagulation performance study of Moringa oleifera cationic protein fractions with varying water hardness. J. Environ. Chem. Eng., 4(4), 4690-4698. https://doi.org/10.1016/j.jece.2016.10.029

Nisha, R., Jegathambal, P., Parameswari, K., Kirupa, K. (2017). Biocompatible water softening system using cationic protein from Moringa oleifera extract. Applied Water Science, 1-9. https://doi.org/10.1007/s13201-017-0591-8

Santos, A., Luz, L., Argolo, A., Teixeira, J., Paiva, P., Coelho, L. (2009). Isolation of seed coagulant Moringa oleifera lectin. Process Biochem., 44(4), 504-508. https://doi.org/10.1016/j.procbio.2009.01.002

Alvez, A., Olivera, M., Guttierres, R., Bergamaso, R., Fernandes, M., Salcedo, A. (2017). Protein fractionation of seeds of Moringa oleifera lam and its application in superficial water treatment. Separation and Purification Technology, 180, 114-124. https://doi.org/10.1016/j.seppur.2017.02.040

Dezfooli, S., Uversky, V., Saleem, M., Baharudin, F., Hitam, S., Bachmann, R. (2016). A simplified method for the purification of an intrinsically disordered coagulant protein from defatted Moringa oleifera seeds. Process Biochemistry, 51(8), 1085-1091. https://doi.org/10.1016/j.procbio.2016.04.021

Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. http://www.academia.edu/download/48656718/227680a020160907-2285-6zp3f3.pdf

Mani, S., Jaya, S., Vadivambal, R. (2007). Optimization of solvent extraction of Moringa (Moringa oleifera) seed kernel oil using response surface methodology. Food and Bioproducts Processing, 85(C4), 328-335. https://doi.org/10.1205/fbp07075

Delange, M., Murillo, M., González, V., Gutiérrez, J. (2014). Composición de ácidos grasos del aceite de las semillas de Moringa oleifera que crece en La Habana, Cuba. Rev. Cubana de Plantas Medicinales, 19(2), 197-204. http://scielo.sld.cu/pdf/pla/v19n2/pla08214.pdf

Coates, J. (2006). Interpretation of Infrared Spectra, A Practical Approach. In: Encyclopedia of Analytical Chemistry. R.A. Meyers (Ed.) Copyright John Wiley & Sons Ltd. https://doi.org/10.1002/9780470027318.a5606

Campos, J., Colina, G., Fernández, N., Torres, G., Sulbarán, B., Ojeda, G. (2003). Caracterización del agente coagulante activo de las semillas de Moringa oleifera mediante HPLC. Boletín del Centro de Investigaciones Biológicas, 37(1), 35-43. http://www.produccioncientificaluz.org/index.php/boletin/article/viewFile/3/3

Ndabigengesere, A., Narasiah, I., Talbot, B. (1995). Active agents and mechanism of coagulation of turbid waters using Moringa oleifera. Water Research, 29(2), 703-710. https://doi.org/10.1016/0043-1354(94)00161-Y

Perez, M. (2007). Diseño y síntesis de péptidos para el diagnóstico en la infección por el virus de hepatitis G (GBV-C/HGV). Tesis de Doctorado. Instituto de Investigación Biomédicas August Pi i Sunyer,. Bacelona, page 129-132. http://digital.csic.es/bitstream/10261/22631/5/Perez_Escoda_5.pdf

Kwaambwa, H., & Maikokera, R. (2008). Infrared and circular dichroism spectroscopic characterisation of secondary structure components of a water treatment coagulant protein extracted from Moringa oleifera seeds. Colloids and Surfaces B, 64(1), 118–125. https://doi.org/10.1016/j.colsurfb.2008.01.014


Refbacks

  • There are currently no refbacks.


Copyright © 2018 Journal of Progressive Research in Modern Physics and ChemistryAll rights reserved.

ISSN: 2456-6438

For any query/support contact us at jprmpceditor@scischolars.com, ssroscischolars@gmail.com,

editorial@scischolars.com