Cargando…

Isolation and purification of beta-lactoglobulin from cow milk

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Aich, Ranjit, Batabyal, Subhasis, Joardar, Siddhartha Narayan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Veterinary World 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4774723/
https://www.ncbi.nlm.nih.gov/pubmed/27047145
http://dx.doi.org/10.14202/vetworld.2015.621-624
_version_ 1782418948184080384
author Aich, Ranjit
Batabyal, Subhasis
Joardar, Siddhartha Narayan
author_facet Aich, Ranjit
Batabyal, Subhasis
Joardar, Siddhartha Narayan
author_sort Aich, Ranjit
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4774723
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Veterinary World
record_format MEDLINE/PubMed
spelling pubmed-47747232016-04-04 Isolation and purification of beta-lactoglobulin from cow milk Aich, Ranjit Batabyal, Subhasis Joardar, Siddhartha Narayan Vet World Research Article 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. Veterinary World 2015-05 2015-05-15 /pmc/articles/PMC4774723/ /pubmed/27047145 http://dx.doi.org/10.14202/vetworld.2015.621-624 Text en Copyright: © The authors. http://creativecommons.org/licenses/by/2.0 This article is an open access article licensed under the terms of the Creative Commons Attributin License (http://creative commons.org/licenses/by/2.0) which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.
spellingShingle Research Article
Aich, Ranjit
Batabyal, Subhasis
Joardar, Siddhartha Narayan
Isolation and purification of beta-lactoglobulin from cow milk
title Isolation and purification of beta-lactoglobulin from cow milk
title_full Isolation and purification of beta-lactoglobulin from cow milk
title_fullStr Isolation and purification of beta-lactoglobulin from cow milk
title_full_unstemmed Isolation and purification of beta-lactoglobulin from cow milk
title_short Isolation and purification of beta-lactoglobulin from cow milk
title_sort isolation and purification of beta-lactoglobulin from cow milk
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4774723/
https://www.ncbi.nlm.nih.gov/pubmed/27047145
http://dx.doi.org/10.14202/vetworld.2015.621-624
work_keys_str_mv AT aichranjit isolationandpurificationofbetalactoglobulinfromcowmilk
AT batabyalsubhasis isolationandpurificationofbetalactoglobulinfromcowmilk
AT joardarsiddharthanarayan isolationandpurificationofbetalactoglobulinfromcowmilk