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A highly selective two-way purification method using liquid chromatography for isolating αS2-casein from goat milk of five different breeds


Citation

Mohsin, Aliah Zannierah and Sukor, Rashidah and Selamat, Jinap and Meor Hussin, Anis Shobirin and Ismail, Intan Hakimah and Jambari, Nuzul Noorahya and Jonet, Mohd Anuar (2020) A highly selective two-way purification method using liquid chromatography for isolating αS2-casein from goat milk of five different breeds. Journal of Chromatography B, 1160. art. no. 122380. pp. 1-9. ISSN 1570-0232; ESSN: 1873-376X

Abstract

The main challenges in the purification of αS2-casein are due to the low quantity in milk and high homology with other casein subunits, i.e., αS1-casein, β-casein, and κ-casein. To overcome these challenges, the aim of this study was to develop a two-step purification to isolate native αS2-casein in goat milk from five different breeds; British Alpine, Jamnapari, Saanen, Shami, and Toggenburg. The first step of the purification was executed by anion-exchange chromatography under optimal elution conditions followed by size exclusion chromatography. Tryptic peptides from in-gel digestion of purified αS2-casein were sequenced and analyzed by LC-ESI-MS/MS. From 1.05 g of whole casein, the highest yield of αS2-casein (6.7 mg/mL) was obtained from Jamnapari and the lowest yield (2.2 mg/mL) was from Saanen. A single band of pure αS2-casein was observed on SDS-PAGE for all breeds. The αS2-casein showed coverage percentage of amino acid sequence from 76.68 to 92.83%. The two-step purification process developed herein was successfully applied for isolating native αS2-casein from goat milk with high purity, which will allow for future in vitro studies to be conducted on this protein.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Food Science and Technology
Faculty of Medicine and Health Science
Institute of Tropical Agriculture and Food Security
DOI Number: https://doi.org/10.1016/j.jchromb.2020.122380
Publisher: Elsevier
Keywords: Goat αS2-casein; Protein identification; Goat breed; Ion-exchange chromatography; Size-exclusion chromatography; LC-ESI-MS/MS
Depositing User: Ms. Nuraida Ibrahim
Date Deposited: 06 Jul 2022 08:18
Last Modified: 06 Jul 2022 08:18
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.jchromb.2020.122380
URI: http://psasir.upm.edu.my/id/eprint/87602
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