Isolation and Characterization of Chickpea (Cicer arietinum L.) Seed Protein Fractions
Proteins of ground chickpea seeds were extracted with sodium hydroxide (NaOH) solution and precipitated with addition of acid (isoelectric precipitate (C-IP)) and by cryoprecipitation (cryoprecipitate (C-CP)). The protein isolates were characterized by Native PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), reversed-phase high performance liquid chromatography (RP-HPLC) and electrospray-ionization mass spectrometry (ESI/MS). Both the isoelectric precipitate and cryoprecipitate contained the globulin protein 11S legumins and 7S vicilins as the major protein fractions and 2S albumin proteins as a minor protein fraction. The major subunits of RP-HPLC protein fractions from both cryoprecipitate and isoelectric precipitate were found to contain subunits of both legumins and vicilins. SDS-PAGE identified legumin α-subunits with MW 40.6 and 39.5 kDa and legumin β- subunits with MW 23.5 and 22.5 kDa, and vicilin subunits with MW 70.2, 50.7, 35.0, 33.6, 18.9 and 15.5 kDa. ESI-MS molecular weights 35,366, 35,268 and 14,648 Da are likely vicilin subunits while the 24,894 Da is a legumin β-subunit.
KeywordsChickpea protein Cryoprecipitate Isoelectric precipitate Legumin Vicilin ESI-MS
The authors acknowledge the assistance from Ms. Beata Usakiewicz with ESI-MS analysis.
- Alli, I., & Ramadan, A. (2003). High Gelling Protein and a Process for Obtaining Same from Soybean. United States Patent No: US 6,569,484 B1.Google Scholar
- Alli, I., Gibbs, B. F., Okoniewska, M. K., Konishi, Y., & Dumas, F. (1994). Identification and characterisation of phaseolin polypeptide subunits in a crystalline food protein isolate from large lima beans (Phaseolus lunatus). Journal of Agricultural and Food Chemistry, 42, 2679–2683.CrossRefGoogle Scholar
- Casey, R., Domoney, C., & Smith, A. M. (1993). Biochemistry and molecular biology of seed products. In R. Casey & D. R. Davies (Eds.), Peas: Genetics, molecular biology and biotechnology (pp. 121–164). CAB International: Wallingford, U.K.Google Scholar
- Chavan, J. K., Kadam, S. S., & Salunkhe, D. K. (1989). Chickpea. In D. K. Salunkhe & S. S. Kadam (Eds.), CRC Handbook of world food legumes: Nutritional chemistry, processing technology, and utilization. Volume I (pp. 247–288). Florida: CRC Press.Google Scholar
- Davis, B. J. (1964). Disc electrophoresis-II. Method and application to human serum proteins. Annals of the New York Academy of Sciences, 12, 404–427.Google Scholar
- Duke, J. A. (1981). Handbook of legumes of world economic importance. Food and agriculture organization of the united nations (pp. 52–57). N.Y: Plenum Press.Google Scholar
- Fan, T. Y., & Sosulski, F. W. (1974). Dispersibility and isolation of protein from legume flours. Canadian Institute of Food Science and Technology Journal, 7, 256–259.Google Scholar
- FAO (2004). Statistical Database of Agriculture Crops. Food and Agriculture Organization, Rome. Available at: www.fao.org. Accessed 15 November 2004.
- Gueguen, J. (1991). Pea and fababean proteins. In B. J. F. Hudson (Ed.), Development in food proteins (pp. 35–78). Elsevier: London, U.K.Google Scholar
- Hulse, J. H. (1991). Nature, composition and utilization of grain legumes (pp. 11-27). In: Uses of tropical legumes: Proceedings of a Consultants' Meeting, 27-30 March 1989, ICRISAT Center. ICRISAT, Patancheru, A.P. 502 324, India.Google Scholar
- Moreira, A. M., Hermodson, M. A., Larkins, B. A., & Nielsen, N. C. (1979). Partial characterization of the acidic and basic polypeptides of glycinin. Journal of Biological Chemistry, 19, 9921–9926.Google Scholar
- Sathe, S. K., Lilley, G. G., Mason, A. C., & Weaver, C. M. (1987). High-resolution sodium dodecyl sulfate polyacrylamide gel electrophoresis of soybean (Glycine max L.) seed proteins. Cereal Chemistry, 64, 380–384.Google Scholar
- Vaidehi, M. P., & Kadam, S. S. (1989). Soybean (pp. 1–31). In D. K. Salunkhe & S. S. Kadam (Eds.), CRC Handbook of world food legume: nutritional chemistry, processing technology, and utilization (Vol. III). Florida: CRC Press.Google Scholar
- Vairinhos, F., & Murray, D. R. (1982). Changes in polypeptides composition during seed development in chickpea (Cicer arietinum L.). Zeitschrift fur Pflanzenernahrung und Bodenkunde, 106, 447–452.Google Scholar