LENTIL SEED ASPARTATE APvIINOTRANSFERASE ISOENZYMES I. ISOLATION and PARTIAL PURIFICATION

Abstract

Three electrophoretically distinct aspartate minotransferase isoenzymes named AAT-1, AAT-2and AAT-3, were separated from lentil (Lens culinaris Medik.) seeds by extraction, ammonium sulphate precipitation, hydroxylapatite and DEAE-cellulose column chromatographies. AAT-1 was purified 228, AAT-2,42 and AAT-3, 3.8 fold. The isoenzymes were examined by means of polyacrylamide gel electrophoresis.Owing to its key role in general metabolism, aspartate adotransferase (AAT, E.C. 2.6.1.1.) which was formerly called glutamic oxaloacetic transaminase (GOT), occurs widely in nature. In animals, cytbsolic and rnitochon+al forms of AAT have been characterized and purified (1-4). AAT has also been studied in various plants (5, 6) and different isoeozyrnic forms have been r&-, ported. The enzyme has been extensively characterized in wheat germ (7,8), cauliflower (9), cotton seeds (lo), pea (ll), soybean (12-14), oat (15), lupine (16) and alfalfa (17,18).

Özet

Elektroforetik balurndan far& ii~ aspartat aminotransferaz izoenzimi yegil mercimek (Lens culinaris Medik.) tohudarmdan ekstraksiyon , amonyum sulfatla goktiirme, hidroksilapatit ve DEAE-seluloz sutun kromatografisi yontemleri kullanllarak aynld~. AAT-1, AAT-2 ve AAT-3 olarak adlandlrilan bu izoenzimler, slras~yla 228, 42 ve 3.8 kez saflagtlrdd~. Izoenzimler poliakrilamid jelelektroforezi ile incelendi.Elektroforetik balurndan far& ii~ aspartat aminotransferaz izoenzimi yegil mercimek (Lens culinaris Medik.) tohudarmdan ekstraksiyon , amonyum sulfatla goktiirme, hidroksilapatit ve DEAE-seluloz sutun kromatografisi yontemleri kullanllarak aynld~. AAT-1, AAT-2 ve AAT-3 olarak adlandlrilan bu izoenzimler, slras~yla 228, 42 ve 3.8 kez saflagtlrdd~. Izoenzimler poliakrilamid jelelektroforezi ile incelendi.