One-step method for isolation and purification of native beta-lactoglobulin from bovine whey
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2012
Authors
Stojadinović, MarijaBurazer, Lidija
Ercili-Cura, Dilek
Sancho, Ana
Buchert, Johanna
Ćirković-Veličković, Tanja
Stanić-Vučinić, Dragana
Article (Published version)
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BACKGROUND: The major whey protein beta-lactoglobulin (BLG) has been widely studied for its functional properties. The aim of this study was to develop an efficient, inexpensive and rapid one-step method for the isolation and purification of BLG while preserving its native structure. RESULTS: BLGwas purified fromdefattedwheyobtainedfromrawcow's milkbyanionexchangechromatography. Protein purity and identitywere determined using reverse phase high-performance liquid chromatography andmass spectrometry. Total BLG yield was 80% with protein purity from 97 to 99%. BLG isoforms A and B were separated into fractions of 91 and 99% purity respectively. The structure and native conformation of the isolated BLGwere compared with those of standard commercial BLG by circular dichroism spectrometry, susceptibility to various crosslinking enzymes and enzyme-linked immunosorbent assay inhibition. CONCLUSION: Theproposedmethodis veryuseful for the rapid preparationofBLGsuitable for studying antigenican...dmolecular characteristics of this protein, aswell as the effect of food processing on these properties. The procedure requires only 1 day for the purification of about 300 mgof BLG from a single run using a small column (2.5 cmx20 cm) of diethylaminoethyl Sephadex and has potential for scaling up. (C) 2011 Society of Chemical Industry
Keywords:
native ss-lactoglobulin / isolation / anion exchange chromatography / purificationSource:
Journal of the Science of Food and Agriculture, 2012, 92, 7, 1432-1440Publisher:
- Wiley-Blackwell, Malden
Funding / projects:
- Molecular properties and modifications of some respiratory and nutritional allergens (RS-MESTD-Basic Research (BR or ON)-172024)
- Reinforcement of the Faculty of Chemistry, University of Belgrade, towards becoming a Center of Excellence in the region of WB for Molecular Biotechnology and Food research (EU-FP7-256716)
Note:
- Peer-reviewed manuscript: http://intor.torlakinstitut.com/handle/123456789/675
Related info:
DOI: 10.1002/jsfa.4722
ISSN: 0022-5142
PubMed: 22083849
WoS: 000302468200015
Scopus: 2-s2.0-84859419583
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TorlakTY - JOUR AU - Stojadinović, Marija AU - Burazer, Lidija AU - Ercili-Cura, Dilek AU - Sancho, Ana AU - Buchert, Johanna AU - Ćirković-Veličković, Tanja AU - Stanić-Vučinić, Dragana PY - 2012 UR - http://intor.torlakinstitut.com/handle/123456789/361 AB - BACKGROUND: The major whey protein beta-lactoglobulin (BLG) has been widely studied for its functional properties. The aim of this study was to develop an efficient, inexpensive and rapid one-step method for the isolation and purification of BLG while preserving its native structure. RESULTS: BLGwas purified fromdefattedwheyobtainedfromrawcow's milkbyanionexchangechromatography. Protein purity and identitywere determined using reverse phase high-performance liquid chromatography andmass spectrometry. Total BLG yield was 80% with protein purity from 97 to 99%. BLG isoforms A and B were separated into fractions of 91 and 99% purity respectively. The structure and native conformation of the isolated BLGwere compared with those of standard commercial BLG by circular dichroism spectrometry, susceptibility to various crosslinking enzymes and enzyme-linked immunosorbent assay inhibition. CONCLUSION: Theproposedmethodis veryuseful for the rapid preparationofBLGsuitable for studying antigenicandmolecular characteristics of this protein, aswell as the effect of food processing on these properties. The procedure requires only 1 day for the purification of about 300 mgof BLG from a single run using a small column (2.5 cmx20 cm) of diethylaminoethyl Sephadex and has potential for scaling up. (C) 2011 Society of Chemical Industry PB - Wiley-Blackwell, Malden T2 - Journal of the Science of Food and Agriculture T1 - One-step method for isolation and purification of native beta-lactoglobulin from bovine whey EP - 1440 IS - 7 SP - 1432 VL - 92 DO - 10.1002/jsfa.4722 ER -
@article{ author = "Stojadinović, Marija and Burazer, Lidija and Ercili-Cura, Dilek and Sancho, Ana and Buchert, Johanna and Ćirković-Veličković, Tanja and Stanić-Vučinić, Dragana", year = "2012", abstract = "BACKGROUND: The major whey protein beta-lactoglobulin (BLG) has been widely studied for its functional properties. The aim of this study was to develop an efficient, inexpensive and rapid one-step method for the isolation and purification of BLG while preserving its native structure. RESULTS: BLGwas purified fromdefattedwheyobtainedfromrawcow's milkbyanionexchangechromatography. Protein purity and identitywere determined using reverse phase high-performance liquid chromatography andmass spectrometry. Total BLG yield was 80% with protein purity from 97 to 99%. BLG isoforms A and B were separated into fractions of 91 and 99% purity respectively. The structure and native conformation of the isolated BLGwere compared with those of standard commercial BLG by circular dichroism spectrometry, susceptibility to various crosslinking enzymes and enzyme-linked immunosorbent assay inhibition. CONCLUSION: Theproposedmethodis veryuseful for the rapid preparationofBLGsuitable for studying antigenicandmolecular characteristics of this protein, aswell as the effect of food processing on these properties. The procedure requires only 1 day for the purification of about 300 mgof BLG from a single run using a small column (2.5 cmx20 cm) of diethylaminoethyl Sephadex and has potential for scaling up. (C) 2011 Society of Chemical Industry", publisher = "Wiley-Blackwell, Malden", journal = "Journal of the Science of Food and Agriculture", title = "One-step method for isolation and purification of native beta-lactoglobulin from bovine whey", pages = "1440-1432", number = "7", volume = "92", doi = "10.1002/jsfa.4722" }
Stojadinović, M., Burazer, L., Ercili-Cura, D., Sancho, A., Buchert, J., Ćirković-Veličković, T.,& Stanić-Vučinić, D.. (2012). One-step method for isolation and purification of native beta-lactoglobulin from bovine whey. in Journal of the Science of Food and Agriculture Wiley-Blackwell, Malden., 92(7), 1432-1440. https://doi.org/10.1002/jsfa.4722
Stojadinović M, Burazer L, Ercili-Cura D, Sancho A, Buchert J, Ćirković-Veličković T, Stanić-Vučinić D. One-step method for isolation and purification of native beta-lactoglobulin from bovine whey. in Journal of the Science of Food and Agriculture. 2012;92(7):1432-1440. doi:10.1002/jsfa.4722 .
Stojadinović, Marija, Burazer, Lidija, Ercili-Cura, Dilek, Sancho, Ana, Buchert, Johanna, Ćirković-Veličković, Tanja, Stanić-Vučinić, Dragana, "One-step method for isolation and purification of native beta-lactoglobulin from bovine whey" in Journal of the Science of Food and Agriculture, 92, no. 7 (2012):1432-1440, https://doi.org/10.1002/jsfa.4722 . .