Extraction and Purification of Lactoferrin from Camel Milk and Investigation of Its Amylase Activity

 Background and purpose: Lactoferrin is found in mucus, milk, and colostrum secretions and has antimicrobial activities, improves iron absorption, and enhances immune responses. Lactoferrin has the ability to degrade starch. The aim of this study was to investigate the preservation of milk lactoferrin enzymatic activity after purification by ion exchange chromatography. Materials and methods: In this experimental research, fat and casein were removed from camel milk and lactoferrin was purified by ion exchange chromatography using CM-Sephadex-C-50. Km and Vmax values of lactoferrin amylase activity, pH, and optimum reaction temperature were determined. For this purpose, 0.5% solution of cornstarch was used as a substrate and iodine activity test was used to determine the unused amounts. The optimum temperature for amylase activity was determined by paper chromatography. Results: The presence of lactoferrin was seen at about 73 KD and its average concentration was 60 µg/ml. Km and Vmax values were 2mg.ml-1 and 0.2 mg.ml-1.min-1, respectively. Lactoferrin also showed extensive amylase activity in pH 4-7 and had the highest activity at pH 5. The amylase activity of lactoferrin reached its maximum at 40°C. Enzymatic digestion caused hydrolysis of starch and its conversion to maltose and glucose. Conclusion: According to the present study, ion exchange chromatography could isolate high purity lactoferrin. High amylase activity of lactoferrin showed that its protein structure was preserved after passing through the chromatographic column.