SIMPLE METHOD FOR PURIFICATION OF WHEAT GERM LECTIN BY AFFINITY CHROMATOGRAPHY

A.Taha, Ali; O. Muhynddin, Mohamed; M. AI-Aubadi, Inas

	SIMPLE METHOD FOR PURIFICATION OF WHEAT GERM LECTIN BY AFFINITY CHROMATOGRAPHY
The Iraqi Journal of Agricultural Science
Article 1, Volume 39, Issue 2, 2008, Pages 44-53
Authors
Ali A.Taha; Mohamed O. Muhynddin; Inas M. AI-Aubadi
Abstract
Because of various application of purified lectin ,the aim of this study was to extract it from wheat germ which is one of the byproducts from wheat processing. The extraction steps including fat elimination from wheat germ by heptan fellowed by lectin extraction by 0.05 M hydrochloric acid solution in ratio (1: 10)(w/v) for one hour at 25C . The crud extract was subjected to different purification steps including concentration by ultrafiltration by this step it was possible to eliminate high portion of low molecular wight compounds which is below 10000 dalton. The difiltration was done to concentrated the sample by 0.01 M Tris-HCL, pH 8.5 .The purification folds and yeild of the lectin were 5.71 and 98.34% respectively. Ion-exchange chromatography by bachwise procedure using Diethel amino ethel cellulose (DEAE¬Cellulose) then affinity chromatography by chitin column, where the chitin itself was prepared from outer shells of shrimp ,we obtained about 40 g chitin which is equel to 20% chitin from the dry shells then the chitin was used as a matrix and a ligand in affinity chromatography for purification of the lectin after this step the purification fold was 20 and yeild of the lectin was 19.51 % . The lectin was purified to homogeneity as indicated by presence of one band in polyacrylamide gel electrophoresis under denaturing conditions in presence and absence of the reducing agent 2-mercaptoethanol .The amount of lectin is about 60mg /1000 gm of wheat germ when using the steps which indicated in this study .

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