Purification of Lactoperoxidase Enzyme from Camel's Milk and Survey Its Antibacterial Effects against Pseudomonas aeru¬ginosa and Clostridium septicom

Lactoperoxidase is an enzyme of the oxidoreductase family. Lactoperoxidase is an important antimicrobial agent. Applications of lactoperoxidase are being found as a preservative in food and cosmetics. Immobilized LPO provides several significant benefits such as، easily separated from the reaction products، reducing production costs by efficient recycling and control of the process. Camel lactoperoxidase (LPO) was purified using CM sephadex C-50 ion-exchange chromatography، and sephadex G-100 gel filtration from camel milk. The activity of LPO was measured by using TMB (3، 3، 5، 5-tetramethylbenzidine) as a choromogenic substrate at pH6. 0. Purification degree was controlled with SDS-PAGE. The number of bacteria were found by spread plate technique and used 10 -6 dilution per ml for accomplishment of tests. According to statistical manner 3 test groups and one control group by four replications with equal conditions were formed for each bacterium. Test groups include 2 reagent groups (H2O2، tiocyanate) and one group containing H2O2 and tiocyanate together and one complete system {H2O2 (0. 03Mm)، tiocyanate (1Mm)، LPO (0. 02µg/ml)}. The numbers of bacteria were counted at 0، 60 and 360 mints by using spread plate technique. According to two-way variance analysis، significant results were shown between the effects of complete system and other for antibacterial activity (P<0. 0001). The results shown 46% reduced of colony form unit for Pseudomonas aeru ginosa and 100% reduced of colony form unit for Clostridium septicum after 360 Minutes. As a result، Lactoperoxidas enzyme was extracted from Camel Milk shown significantly anti-bacterial activity on Pseudomonas aeru ginosa and Clostridium septicum.