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THE PURIFICATION OF VICILIN AND ITS SUBUNITS FROM PEA SEEDS (PISUM SATIVUM L.)

ZHAO WEN-MING D.BOULTER,J.A.GATEHOUSE (Northwestern College of Agriculture) (University of Durham,U.K.)  
The purification of vicilin andits subunits from pea seeds is descri-bed in detail.Pea seed meal wasextracted with borate buffer and theextract was made 85% saturation with(NH_4)_2 SO_4.The material precipi-tated was collected by centrifugationand discarded;the supernatant,whichcontained vicilin,was dialysed andlyophilised and then the vicilin waspurified by gel filtration.The vici-lin subunits were separated and puri-fied by ion-exchange chromatographyon DEAE-cellulose (in 8M urea asdenaturant) and by gel-filtration onsephacryl S-200 (in 70 % formic acidas denaturant).Analysis by gel elec-trophoresis showed that these me-thods gave preparation of purified vi-cilin and its separated componentsubunits.The results also showedthat at pH 8.5 the subunit of Mr33,000 has the least negative chargeand the 12,000 Mr subunit has themost negative charge.ACKNOWLEDGEMENTS We thank Mr.Dave Bown for in-valuable technical advice and assis-tance.We also thank Drs.R.R.D.Croy,Dr.M.Evans and Dr.J.N.Yarwood for their suggestions andhelp.This work was supported byfinancial aid from Department of Bot-any,University of Durham.U.K.
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