Determination of Albumins, Globulins, Prolamins, and Glutelins in All Varieties of Phaseolus Beans.
The extractants water, NaCl, ethanol, and NaOH were applied sequentially to allow the separation of albumins, globulins, prolamins, and glutelins, respectively .
A decrease was found for the majority of the Phaseolus bean varieties, particularly for common beans (albumins, globulins, prolamins, and glutelins).
It was found that this fraction is formed by albumins, globulins, prolamins, and glutelins, which have been also reported in plants such as amaranth.
M, molecular weight marker; TP, total protein; ALB, albumins; 7S GLB, 7S globulins; 11S GLB, 11S globulins; PRO, prolamins; GLU, glutelins.
Nut protein sequences for analysis in silico are as follows: Glutelin (44aa, superfamily Glutelin, GenBank accession: AAC69515.1), 11S legumin (505aa, superfamily Globulin, GenBank accession: ABW86979.1), 7S vicilin (784aa, superfamily Globulin, GenBank accession: ABV49593.1), 7S vicilin (792aa, superfamily Globulin, GenBank accession: ABV49592.1), putative allergen I1 (143aa, superfamily Albumin, GenBank accession: AAO32314.1), and putative 7S vicilin (102aa, superfamily Globulin, GenBank accession: AAZ93628.1).
The glutelin fraction is particularly difficult to characterize because its solubility is restricted to acids and bases.
The glutelin fraction remaining in the meal was then extracted with 0.05 M NaOH for 30 min.
Following this, the nitrocellulose membrane was incubated with polyclonal antibodies specific for rice glutelin (Krishnan et al., 1986), globulin (Krishnan et al., 1992), prolamin (Krishnan and White, 1995), maize binding protein (Boston et al., 1991), and elongation factor lA (Habben et al., 1995) that were diluted 1:500 in TBS containing 50 g [kg.sup.-1] non-fat dairy milk for 1 h at room temperature.
The rice glutelin, the most abundant storage protein of rice endosperm (Krishnan and Okita, 1986), is composed of two major subunits of 34 to 37 kDa ([Alpha]-subunit) and 21 to 22 kDa ([Beta]-subunit).
Glutelins, the predominant storage proteins of rice endosperm, have also been reported to occur in embryos (Komatsu et al., 1993).
In this study, we used antibodies raised against purified rice endosperm globulins and against glutelins and have demonstrated that similar proteins accumulate within protein bodies in the embryo.