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Isolation, characterization and expression analysis of a nutritionally enhanced α-prolamin gene and protein during developing spikes of finger millet (Eleusine coracana)

Published online by Cambridge University Press:  14 September 2017

Lalan Kumar
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
Dinesh Pandey
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
Anil Kumar*
Affiliation:
G B Pant University of Agriculture and Technology, Pantagar, Uttarakhand, India
*
*Correspondence Email: anilkumar.mbge@gmail.com

Abstract

In the present study a combination of BLAST mediated homology search and 3′ RACE was utilized to isolate the full-length gene of Eleusine coracana alpha prolamin (Ec-α-prolamin) from finger millet. Phylogenetic analysis of Ec-α-prolamin along with related prolamin genes of different cereals and millets shows the clustering of Ec-α-prolamin in a separate group. Secondary structure prediction reveals 59.4% alpha helix structure, a structural hallmark of Ec-α-prolamin. Besides this, the protein also possesses a balanced proportion of all essential amino acids. Expression analysis based on qPCR shows increased accumulation of α-prolamin transcripts in developing grains of finger millet until attainment of seed maturity. Western blotting, using a monospecific anti-α-prolamin antibody, further confirmed the expression of a 22 kDa band in the S3 and S4 stages of developing spikes. The heterologous expression of isolated full-length Ec-α-prolamin could be potentially harnessed for making nutritionally enhanced functional food products and value-added industrial products.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2017 

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References

Arcalis, E., Stadlmann, J., Marcel, S., Drakakaki, G., Winter, V., Rodriguez, J., Fischer, R., Altmann, F. and Stoger, E. (2010) The changing fate of a secretory glycoprotein in developing maize endosperm. Plant Physiology 153, 693702.Google Scholar
Bradford, M.M. (1976) A rapid and sensitive method for the quantification of micrograms quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 1723, 248254.Google Scholar
Byaruhanga, Y.B., Emmambux, M.N., Belton, P.S., Wellner, N., Ng, K.G. and Taylor, J.R.N. (2006) Alteration of kafirin and kafirin film structure by heating with microwave energy and tannin complexation. Journal of Agricultural and Food Chemistry 54, 41984207.Google Scholar
Charbonnier, L., Terce-Laforgue, T. and Mosse, J. (1981) Rye proalmin: extractability, separation and characterisation. Journal of Agricultural and Food Chemistry 29, 968973.Google Scholar
Ciccocioppo, R., Di Sabatino, A. and Corazza, G.R. (2005) The immune recognition of gluten in celiac disease. Clinical and Experimental Immunology 140, 408416.Google Scholar
da Silva, L.S. and Taylor, J.R.N. (2005) Physical, mechanical, and barrier properties of kafirin films from red and white sorghum milling fractions. Cereal Chemistry 82, 914.Google Scholar
Dida, M.M., Srinivasachary, , Ramakrishnan, S., Bennetzen, J.L., Gale, M.D. and Devos, K.M. (2007) The genetic map of finger millet, Eleusine coracana . Theoretical and Applied Genetics 114, 321332.Google Scholar
Food and Agriculture Organization of the United Nations (FAO) (1970) Amino-acid Content of Foods and Biological Data on Proteins. Rome: FAO.Google Scholar
Forde, B.G., Heyworth, A., Pywell, J. and Kreis, M. (1985) Nucleotide sequence of a B1 hordein gene and the identification of possible upstream regulatory elements in endosperm storage protein genes from barley, wheat and maize. Nucleic Acids Research 13, 73277339.Google Scholar
Guruprasad, K., Reddy, B.V.P. and Pandit, M.W. (1990) Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Engineering 4, 155164.Google Scholar
Hoffman, P. (2009) New grain testing procedure available. Agri-View. Madison,WI: University of Wisconsin.Google Scholar
Indira, R. and Naik, M.S. (1971) Nutrient composition and protein quality of some minor millets. Indian Journal of Agriculture Sciences 41, 795797.Google Scholar
Jayaram, B., Dhingra, P., Lakhanib, B. and Shekhar, S. (2012) Targeting the near impossible: pushing the frontiers of atomic models for protein tertiary structure prediction. Journal of Chemical Sciences 124, 8391.Google Scholar
Letunic, I., Doerks, T. and Bork, P. (2012) Recent updates to the protein domain annotation resource. Nucleic Acids Research 40, 302305.Google Scholar
McDonough, Cassandra M., Lloyd, W.R. and Serna-Saldivar, S.O. (2000) The Millets: Handbook of Cereal Science and Technology. Boca Raton, CRC Press.Google Scholar
Mertz, E.T., Bates, L.S. and Nelson, O.E. (1964) Mutant gene that changes protein composition and increases lysine content of maize endosperm. Science 145, 279280.CrossRefGoogle ScholarPubMed
Mills, E.N., Jenkins, J.A., Alcocer, M.J. and Shewry, P.R. (2004). Structural, biological, and evolutionary relationships of plant food allergens sensitizing via the gastrointestinal tract. Critical Review Food Science Nutrition 44, 379407.CrossRefGoogle ScholarPubMed
Müller, M. and Knudsen, S. (1993) The nitrogen response of a barley C-hordein promoter is controlled by positive and negative regulation of the GCN4 and endosperm box. The Plant Journal 4, 343355.CrossRefGoogle ScholarPubMed
Muthamilarasan, M., Suesh, B.V., Pandey, G., Kumari, K., Parid, S.K. and Prasad, M. (2014) Development of 5123 intron-length polymorphic markers for large-scale genotyping applications in foxtail millet. DNA Research 21, 4152.Google Scholar
Osborne, T.B. (1924). The Vegetable Proteins, 2nd edition. London: Longmans, Green and Co.Google Scholar
Ravindran, G. (1992). Seed proteins of millets: amino acid composition, proteinase inhibitors and in vitro digestibility. Food Chemistry 44, 1317.Google Scholar
Riebel, M.J. and Mankato, M.N. (2005) Biopolymer including Prolamin and methods of making it. Patent US 20060155012 A1.Google Scholar
Mandal, S. and Mandal, R.K. (2000). Seed storage proteins and approaches for improvement of their nutritional quality by genetic engineering. Current. Science 79, 576589.Google Scholar
Serna-Saldivar, S. and Rooney, L.W. (1995) Structure and chemistry of sorghum and millet, pp. 69124 in Dendy, D.A.V. (ed), Sorghum and Millets: Chemistry and Technology. St Paul, MN, USA: American Association of Cereal Chemists.Google Scholar
Shewry, P.R. (2002) The seed storage proteins of pseudocereals and minor cereals, pp 124 in Belton, P. and Taylor, J. (eds), Pseudocereals and Less Well-Known Cereals. New York: Springer-Verlag.Google Scholar
Shobana, S., Sreerama, Y.N. and Malleshi, N.G. (2009) Composition and enzyme inhibitory properties of finger millet (Eleusine coracana L.) seed coat phenolics: mode of inhibition of α glucosidase and α amylase. Food Chemistry 115, 12681273.Google Scholar
Sørensen, M.B., Cameron-Mills, V. and Brandt, A. (1989). Transcriptional and post transcriptional regulation of gene expression in developing barley endosperm. Molecular and General Genetics 217, 195201.CrossRefGoogle Scholar
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. and Kumar, S. (2011) MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology Evolution 28, 27312739.Google Scholar
Tatham, A.S., Fido, R.J., Moore, C.M., Kasarda, D.D., Kuzmicky, D.D., Keen, J.N. and Shewry, P.R. (1996) Characterisation of the major prolamin of Tef (Eragrostis tef) and finger millet (Eleusine coracana). Journal of Cereal. Science 24, 6571.Google Scholar
Taylor, J., Taylor, J.R.N., Dutton, M.F. and De Kock, S. (2005). Identification of kafirin film casting solvents. Food Chemistry 90, 401408.CrossRefGoogle Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.Google Scholar
Upadhyaya, H.D., Gowda, C.L.L. and Reddy, G.V. (2007) Morphological diversity in finger millet germplasm introduce from Southern and Eastern Africa. ICRISAT.Google Scholar
van Eckert, R., Bond, J., Rawson, P., Klein, Ch.L., Stern, M. and Jordan, T.W. (2010) Reactivity of gluten detecting monoclonal antibodies to a gliadin reference material. Journal of Cereal Science 51, 198204.Google Scholar
Venkateswaran, V. and Vijayalakshmi, G. (2010) Finger millet (Eleusine coracana) – an economically viable source for anti-hypercholesterolemic metabolites production by Monascus purpureus . Journal of Food Science and Technology 47, 426431.Google Scholar
Yoon, U.H., Lee, J.H., Hahn, J.H., Kim, Y.K., Lee, G.S., Ji, H.S., Kim, C.K., Mun, J.H., Kim, Y.M., and Kim, T.H. (2012). Structural and expression analysis of prolamin genes in Oryza sativa L. Plant Biotechnology Report 6, 251262.Google Scholar
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