Phone : 0092 300 7786573

Advances in Animal and Veterinary Sciences

Short Communication
Adv. Anim. Vet. Sci. 2 (2): 78 - 80
http://dx.doi.org/10.14737/journal.aavs/2014/2.2.78.80
View Full HTML
Download PDF

Ranjit Aich1, Subhasis Batabyal2, Siddhartha Narayan Joardar2*
1College of Veterinary Science and Animal Husbandry, Mhow–453 446, Madhya Pradesh, India; 2West Bengal University of Animal and Fishery Sciences, Belgachia, Kolkata–700 037, West Bengal, India
*Corresponding author: joardar69@gmail.com

ABSTRACT
It has been observed that 82 percent of milk–allergic patients are sensitive to beta–lactoglobulin (β–lg), a major milk protein that accounts for approximately 10 to 15 percent of total milk proteins. The modification in β–lg is considered a promising venture to mitigate milk allergies. With the aim of standardizing a convenient method for isolation and purification of β–lg from buffalo milk, the present study was designed to keep its antigenicity intact, so that, the purified β–lg can be used to detect buffalo milk protein intolerance. Raw milk was collected from Murrah breed of buffalo from Haringhata farm (West Bengal) and converted to skimmed milk by removing fat globules. Casein protein was removed by acidification to pH 4.6 by adding 3 M HCl. The β–lg was isolated by gel filtration chromatography using Sephacryl S–200 from supernatant whey protein fraction. Further, β–lg was purified by anion–exchange chromatography using DEAE–Sepharose. Molecular weight of the purified buffalo β–lg was 18.05kDa as assessed by the gel documentation system using standard molecular weight marker (range 14.3 to 97.4 kDa) in 15 percent one–dimensional SDS–PAGE. The isolated β–lg was almost in pure form as the molecular weight of purified β–lg monomer is 18kDa. The study revealed a simple and suitable method for isolation of β–lg from whey protein in pure form that might be exploited for any basic and applied studies related to buffalo milk protein intolerance.

Key Words: Beta–lactoglobulin, Buffalo milk, Column chromatography, Milk protein allergy, SDS–PAGE