COMPARATIVE ELECTROPHORETIC PATTERNS OF ALBUMINS/ GLOBULINS EXTRACTED FROM DRY GRAINS AND GREEN MALTS OF BARLEY VARIETIES

Abstract

Ispitana je mogućnost primjene različitih elektroforetskih metoda razdvajanja albumina/globulina slijeđenih nespecifičnim bojanjem proteina, te specifičnom detekcijom glikoproteina i aminopeptidaza za razlikovanje sorti ječma. Albumini/globulini ekstrahirani iz suhoga zrna i zelenoga slada šest sorti ječma razdijeljeni su poliakrilamid gel elektroforezom nativnih uzoraka (10 % T, pH=8,9), poliakrilamid gel elektroforezom u prisutnosti natrijeva dodecil-sulfata te izoelektričnim fokusiranjem u gradijentima pH 3,5-9,5 i pH 4,0-6,5. Ekstrakti proteina iz suhoga zrna davali su bolje rezultate od onih iz zelenoga slada. Rezultati upućuju da bi razdjeljivanje albumina/globulina pomoću SDS-PAGE i izoelektričnoga fokusiranja u gradijentu pH 3,5-9,5 slijeđeno bojanjem proteina pomoću Coomassie Blue moglo poslužiti za razlikovanje hrvatskih sorti ječma. Sortu Angora moguće je, nedvojbeno, razlikovati od ostalih sorti koje se mogu grupirati u skupine kako slijedi: Rodnik/Sladoran, Barun/Rex te sorta Martin. Bojanje glikoproteina ne osigurava razlikovanje ispitanih sorti. Detekcija aktivnosti aminopeptidaze široke specifičnosti, aminopeptidaze koja preferira fenilalanin i leucin, te arginil-aminopeptidaze po provedenom elektroforetskom razdvajanju nije se pokazala pogodnom metodom za razlikovanje ispitanih sorti ječma.Applicability of electrophoretic separations of albumins/globulins, followed by nonspecific protein staining and specific glycoprotein and aminopeptidase detection has been examined for barley variety discrimination. Albumins/globulins extracted from dry grains and green malt of six barley varieties were separated by polyacrylamide gel electrophoresis (10 % T, pH=8.9) of native and sodium dodecyl sulfate denatured proteins, and by isoelectric focusing in pH gradient of 3.5-9.5 and 4.0-6.5. Analysis of dry grain extracts gave better results than green malt extracts. Obtained data indicate that SDS-PAGE of albumins/globulins and IEF in pH gradient 3.5-9.5 followed by Coomassie Blue staining could be useful in Croatian barley variety discrimination. Angora barley could be clearly distinguished from the other varieties which were grouped as follows: Rodnik/Sladoran, Barun/Rex, and Martin. Glycoprotein patterns did not improve the recognition of individual varieties. Broad specificity, phenylalanine and leucine preferring, and arginine specific aminopeptidase were not found as applicable markers for discrimination of examined barley varieties