Isolation and Characterization of Chickpea (Cicer arietinum L.) Seed Protein Fractions

Abstract

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.

Enzymatic Production, Bioactivity, and Bitterness of Chickpea (Cicer arietinum) Peptides

“…Table 4 shows reported peptide sequences from the enzymatic hydrolysis of chickpea proteins. The majority of sequenced chickpea peptides are those produced from chickpea legumin, which is not surprising because legumin is the most abundant storage protein found in chickpeas (Chang, Alli, Molina, Konishi, & Boye, 2012). Sequenced peptides produced through enzymatic hydrolysis have been obtained through simulated gastrointestinal digestion with pepsin followed by pancreatin or by Properties of chickpea peptides .…”

Physicochemical, functional, thermal and structural properties of isolated Kabuli chickpea proteins as affected by processing approaches

“…SDS-PAGE of the protein isolates showed a distribution of molecular weights (MW) ranging from~9 tõ 80 kDa (Fig 4). Chang et al (2012) studied protein isolates from raw chickpea seeds and identified bands of~9-10,~22-24 and~39-41 kDa as legumin subunit 11S protein and those of~15,~18,~33-35 and 70 kDa as vicilin subunit 7S protein. The banding pattern of protein isolates from soaked and raw seeds was similar.…”

“…Chickpeas are a good source of dietary protein, and its protein quality was reported to be better than other legumes Kaur & Singh, 2007). Chickpea protein is mainly composed of globulins (~56%), glutelins (~18%), albumins (~12%), prolamin (~3%) and residual proteins, and the relatively high glutelin content makes chickpea unique compared to other legumes (Chang et al, 2012). Generally, chickpeas are classified into either 'Desi' or 'Kabuli' types based primarily on seed colour and size.…”

Preparation of mayonnaise from extracted plant protein isolates of chickpea, broad bean and lupin flour: chemical, physiochemical, nutritional and therapeutic properties

“…Seven major protein subunits of legumin (11S) corresponding to 22.3, 24, 25.8, 37.5, 39.8, 43.5 and 43.5 kDa, and two major protein subunits of vicilin (7S) corresponding to molecular weights of 33.8 and 50.7 kDa were observed in protein isolates from lupin flour (LPPI). Previous SDS-PAGE results have shown a-subunits of legumin with molecular weight of 40.6 and 39.5 kDa and b-subunits of legumin with molecular weights of 23.5 and 22.5 kDa, and vicilin subunits with molecular weights of 70.2, 50.7, 35, 33.6, 18.9 and 15.5 kDa (Chang et al 2012;Wang et al 2010). Lupin proteins consist from three globulin proteins including 12 protein subunits of b-conglutin (7S), 4 protein subunits of a-conglutin (11S) and a minor c-conglutin (2S) of 170, 315 and 17 kDa molecular weight (Chapleau and de Lamballerie-Anton 2003).…”

“…SDS-PAGE of CPPI demonstrated four major protein subunits of legumin (11S), which correspond to molecular weights of 22.5, 22.8, 23 and 24.1 kDa and seven major protein subunits of vicilin (7S) with molecular weight of 33.7, 34.5, 37.3, 53.4, 58.4, 59.1 and 71.1 kDa. Wang et al (2010) and Chang et al (2012) have found that the globulin protein 11S (legumins) and 7S (vicilins) were the predominant major protein subunits in CPPI using isoelectric and cryoprecipitation techniques, while the 2S albumin proteins was the minor protein subunits CPPI using isoelectric and cryoprecipitation techniques. BBPI showed four major protein subunits of legumin (11S) with molecular weights of 22.4, 22.8, 23.5 and 24.2 kDa and five major protein subunits of vicilin (7S) corresponding to molecular weights of 50.7, 53.5, 55.1, 55.7 and 69.1 kDa.…”