Extraction, Purification, Characterization and Immobilization of Chicken Pepsin and Using it in Protein hydrolysate

K. Issa, Zena; A. Jasim, Munir

doi:10.33762/bagrs.2013.111632

	Extraction, Purification, Characterization and Immobilization of Chicken Pepsin and Using it in Protein hydrolysate
Basrah Journal of Agricultural Sciences
Article 13, Volume 26, Issue 2, 2013, Pages 110-123 PDF (0 K)
DOI: 10.33762/bagrs.2013.111632
Authors
Zena K. Issa; Munir A. Jasim
Abstract
The present study aimed to Isolate pepsin enzyme Ec: 3.4.23.1 from some animal sources and Purified it and Studied it’s Characteristics and Immobilized it as well as it’s practical applications in food industry. The enzyme was extracted from the stomach of three animal sources (Sheep, Chicken and Nuwaibi fish) using five extraction solutions including Distilled water , Sodium chloride (10%) solution , Sodium chloride (6%) Boric acid (2%) solution ,Sodium Phosphate 0.1M and an pH 7.3 solution and Tris-acetic acid 0.4M and an pH 7.6 solution in order to find out the best source of enzyme and the best extraction solution . Sheep's stomach was the best source of enzyme , Sodium chloride (6%) Boric acid (2%) solution was the best extraction solution which gave the highest specific activity 14.3 unit / mg .Protein content for the crude enzyme extracts were con centrated using saturated ammonium sulfate in arrange 30-70% , 20-90% and 20-60% for Sheep, Chicken and Nuwaibi fish pepsin respectively, Dialysis was done using distilled water. Dialyzed Chicken pepsin possesses the highest proteolytic activity. The dialyzed chicken pepsin Immobilized study with four carriers including Silica gel, SRF Resinous, Calicium Alginate and Agar showed that the best carrier was agar which gave the maximum Immobilized efficiency 67%.The best optimum concentration for Immobilize dialyzed chicken pepsin with agar was 3% with the highest proteolytic enzyme 75%. The purified sheep pepsin was Immobilized with agar in 3% concentration .The remainder enzyme activity for agar Immobilized pure Sheep pepsin and dialyzed Chicken pepsin were 25% , 21% respectively after 7 times using and the remainder enzyme activity for agar Immobilize pure Sheep pepsin and dialyzed Chicken pepsin wer 22%, 27% respectively was stored for 60 days in 4 °C. The optimal pH for proteolytic activity for pure free and Immobilize dialyzed Chicken pepsin was 2 .The optimal pH for the stability of free and Immobilize dialyzed Chicken pepsin are 3. The free and Immobilize enzymes lost their activities at alkaline pH values 8 and 9. The optimal temperature for free dialyzed Chicken pepsin was 45 ◌C and 55 °C for its Immobilize form. The optimal temperature for clotting activity for free and Immobilize dialyzed chicken pepsin was 35°C .The optimal temperature for free and Immobilize dialyzed Chicken pepsin stability was 15-45 ◌C. Both free and Immobilize dialyzed Chicken pepsin lost their activities completely at 85°C. Agar Immobilized dialyzed Chicken pepsin was utilized in the preparation of protein concentrate reached the maximum hydrolysis in 180 min. Immobilize Chicken pepsin hydrolysis was approximately similar to free Chicken pepsin which reached 2.6, 2.8 respectively.
Keywords
Extraction Pepsin; Purification Pepsin; Characterization Pepsin; Immobilization Pepsin

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