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R esearch
(Published online:
15-05-2015)
11.
Isolation and purification of beta-lactoglobulin
from cow milk - Ranjit Aich, Subhasis Batabyal and
Siddhartha Narayan Joardar
Veterinary World, 8(5): 621-624
doi:
10.14202/vetworld.2015.621-624
Ranjit Aich:
Department of Veterinary Biochemistry, Faculty of Veterinary and
Animal Sciences, West Bengal University of Animal and Fishery
Sciences, Belgachia - 700 037, Kolkata, India; Department of
Veterinary Biochemistry, College of Veterinary Science and Animal
Husbandry, Mhow - 453 446, Madhya Pradesh, India;
aich80@gmail.com
Subhasis Batabyal:
Department of Veterinary Biochemistry, Faculty of Veterinary and
Animal Sciences, West Bengal University of Animal and Fishery
Sciences, Belgachia - 700 037, Kolkata, India;
batabyals2009@gmail.com
Siddhartha Narayan Joardar: Department of Veterinary
Microbiology, Faculty of Veterinary and Animal Sciences, West
Bengal University of Animal and Fishery Sciences, Belgachia - 700
037, Kolkata, India;
joardar69@gmail.com
Received: 04-01-2015, Revised: 10-04-2015, Accepted: 16-04-2015,
Published online: 15-05-2015
Corresponding author:
Siddhartha Narayan Joardar, e-mail: joardar69@gmail.com
Citation:
Aich R, Batabyal S,
Joardar SN (2015) Isolation and purification of beta-lactoglobulin
from cow milk, Veterinary World, 8(5):621-624.
Abstract
Aim:
The present study was
undertaken to standardize a convenient method for isolation and
purification of β-lactoglobulin (β-lg) from cow milk keeping its
antigenicity intact, so that the purified β-lg can be used for
detection of cow milk protein intolerance (CMPI).
Materials and Methods: Raw
milk was collected from Gir breed of cattle reared in Haringhata
Farm, West Bengal. Milk was then converted to skimmed milk by
removing fat globules and casein protein was removed by
acidification to pH 4.6 by adding 3 M HCl. β-lg was isolated by
gel filtration chromatography using Sephacryl S-200 from the
supernatant whey protein fraction. Further, β-lg was purified by
anion-exchange chromatography in diethylaminoethyl-sepharose.
Molecular weight of the purified cattle β-lg was determined by 15
percent one-dimensional sodium dodecyl sulfate-polyacrylamide gel
electrophoresis and was analyzed by gel documentation system using
standard molecular weight marker.
Results: The
molecular weight of the purified cattle β-lg was detected as 17.44
kDa. The isolated β-lg was almost in pure form as the molecular
weight of purified β-lg monomer is 18kDa.
Conclusion:
The study revealed a simple and suitable method for isolation of
β-lg from whey protein in pure form which may be used for
detection of CMPI.
Keywords: beta-lactoglobulin,
ion-exchange chromatography, milk protein intolerance, sodium
dodecyl sulfate polyacrylamide gel electrophoresis, whey protein.
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