Isolation of Lactoferrin from Camel Milk through Fast Protein Liquid Chromatography and its Antagonistic Activity against Escherichia coli and Staphylococcus aureus
Isolation of Lactoferrin from Camel Milk through Fast Protein Liquid Chromatography and its Antagonistic Activity against Escherichia coli and Staphylococcus aureus
Bushra Niaz1,*, Tahir Zahoor1, Muhammad Atif Randhawa1 and Amer Jamil2
ABSTRACT
Lactoferrin, a multifunctional glycoprotein, occurring as awhey constitute in milk secretions of animals and humandisplays a variety of antimicrobial, antioxidant and immunomodulatory as well as a number of other biological functions. Its potential can be exploited in several food applications. Keeping in view the increased demand of natural antimicrobial agents to control prevalence of food borne pathogens in final packaged food products as well as for food preservation/ biocontrol, this study was designed to isolate lactoferrin from milk of camel. Cation exchange resin (CM-Sephadex C-25) was used for the isolation of lactoferrin using the fast protein liquid chromatography. The lactoferrin was further characterized through sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Disc diffusion method was applied for the evaluation of antibacterial activity of lactoferrin. Minimum inhibitory concentration (MIC) of lactoferrin was determined by 96-well plate method using ELISA reader. Results depicted that CM-Sephadex C-25 was a good cation exchanger for recovery of lactoferrin. Camel raw milk produced 2.4 mg/mL of lactoferrin. The lactoferrin isolated from camel milk exhibited promising antibacterial activity against E. coli and Staphylococcus aureus. Concentration of lactoferrin i.e. 4,000 ppm (4 mg/mL) showed the best results against both the pathogens. The E. coli was more susceptible to inhibition than S. aureusin disc diffusion assay. The MIC of lactoferrin was found to be 0.5 mg/mL for E. coli and 0.2 mg/mL for S. aureus.
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August 2017
Vol. 49, Iss. 4, Pages 1151-1546
