Comparison of Extraction Methods Suitable for...

Comparison of Extraction Methods Suitable for Protein Profile of the Under-Utilized Seed: Wood Apple (Limonia acidissima)

Author Name : L. Banupriya, T. Poongodi Vijayakumar

ABSTRACT

The methodical comparison of protein isolation methods is the first critical step for proteomic studies. In this study, five methods of protein extraction from the seeds of the wood apple were compared by examining the quality in the one-dimensional gel electrophoresis (1-DE), obtained by SDS PAGE of the total protein. The five protein extraction methods were: Trichloroacetic acid (TCA)/acetone precipitation, Tris–HCl buffer, Phenol extraction, Ammonium sulfate precipitation and Alcoholic precipitation. The highest precipitation was recorded with 94% on using TCA/acetone method to high extents. Functional properties such as WAC, Fractionation of protein was done to separate albumin, globulin, prolamine and glutelin. Albumin (66%) and globulin (20%) were the predominant fractions in the protein component of the seed. SDS–PAGE of isolates has revealed the presence of major protein with a molecular weight (MW) of 14 to 97 kDa.The results showed that wood apple seed protein could be a good source of protein fortification or supplement for a variety of food products for protein deficient children as well as a potential food ingredient.

Keywords: Wood apple seed, Protein precipitation, SDS-PAGE, Fractionation