Separation and characterization of mares’ milk alpha(s1)-, beta-, kappa-caseins, gamma-casein-like, and proteose peptone component 5-like peptides.
Abstract: The equine alpha(s1)- and beta-caseins (CN) were purified by chromatography on DEAE-cellulose and by reversed-phase HPLC. The alpha(s1)-, beta-, and kappa-CN were characterized either by monodimensional urea-PAGE or sodium dodecylsulfate (SDS)-PAGE or by bidimensional electrophoresis. Kappa-casein was characterized after electrophoresis by glycoprotein-specific staining. To identify alpha(s1)-CN without ambiguity, internal sequences were determined after trypsin or chymosin digestion of purified alpha(s1)-CN. These sequences, that could be estimated to correspond to 62% of the full protein, presented strong identities with regions of alpha(s1)-CN primary structures of other species. In particular, 51, 48, 43, and 40% identities were obtained with corresponding regions of sow, dromedary, cow, and human alpha(s1)-CN, respectively. On the other hand, trace amounts of equine gamma-CN-like and proteose peptone component 5-like peptides were found in the whole CN. They were identified by microsequencing and corresponded to beta-CN peptides generated by plasmin action on the whole CN. The equine alpha(s1), beta-, and kappa-CN were separated by bidimensional electrophoresis in numerous isoelectric variants with apparent isoelectric points distributed between pH 4.4 to 6.3, 4.4 to 5.9, and 3.5 to 5.5, respectively. The beta- and kappa-CN displayed a more acidic character in the mare than in the cow.
Publication Date: 2002-05-23 PubMed ID: 12018413DOI: 10.3168/jds.S0022-0302(02)74126-XGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article focuses on the purification and characterization of various types of caseins (proteins) found in mares’ milk. This includes alpha, beta, and kappa-caseins as well as gamma-casein-like and proteose peptone component 5-like peptides.
Purification of Caseins
- The caseins in mares’ milk, specifically alpha, beta and kappa-caseins, are purified using chromatography on DEAE-cellulose and by reversed-phase high-performance liquid chromatography (HPLC). Chromatography is a method used to separate the components of a mixture, in this case, the different types of caseins.
Characterization of Caseins
- The different types of caseins are then characterized using techniques such as monodimensional urea-PAGE or sodium dodecylsulfate (SDS)-PAGE, bidimensional electrophoresis, and glycoprotein-specific staining (for kappa-casein).
- The characterization methods help in understanding the individual properties of the caseins.
Sequence Identification
- Furthermore, the alpha casein sequence is identified by trypsin or chymosin digestion. This is a technique used to break down protein sequences to help identify their specific structure.
- The researchers found that these casein sequences showed strong identities with sequences of other species’ alpha caseins, with the highest identities found with sow, dromedary, cow, and human caseins.
Existence of Other Peptides
- Trace amounts of gamma-casein-like and proteose peptone component 5-like peptides were also found in the whole casein. These were identified through a process of microsequencing.
Separation of Caseins
- Lastly, the researchers used bidimensional electrophoresis to separate the alpha, beta, and kappa-caseins into numerous isoelectric variants with specific isoelectric points (the pH at which a particular molecule carries no net electrical charge).
- It’s observed that the beta- and kappa-caseins are comparatively more acidic in mares than in cows.
Cite This Article
APA
Egito AS, Miclo L, López C, Adam A, Girardet JM, Gaillard JL.
(2002).
Separation and characterization of mares’ milk alpha(s1)-, beta-, kappa-caseins, gamma-casein-like, and proteose peptone component 5-like peptides.
J Dairy Sci, 85(4), 697-706.
https://doi.org/10.3168/jds.S0022-0302(02)74126-X Publication
Researcher Affiliations
- Laboratoire des BioSciences de l'Aliment, UC885 INRA, Faculté des Sciences et Techniques, Université Henri Poincaré-Nancy 1, Vandoeuvre-lès-Nancy, France.
MeSH Terms
- Amino Acid Sequence
- Animals
- Caseins / chemistry
- Caseins / genetics
- Caseins / isolation & purification
- Chelating Agents / chemistry
- Chelating Agents / isolation & purification
- Chromatography, High Pressure Liquid / veterinary
- Chromatography, Ion Exchange / veterinary
- Electrophoresis, Gel, Two-Dimensional / veterinary
- Electrophoresis, Polyacrylamide Gel / veterinary
- Female
- Horses / genetics
- Hydrogen-Ion Concentration
- Isoelectric Point
- Milk / chemistry
- Molecular Sequence Data
- Peptide Fragments / chemistry
- Sequence Alignment
- Sequence Homology, Amino Acid
- Species Specificity
Citations
This article has been cited 9 times.- Zhang X, Jiang G, Ji C, Fan Z, Ge S, Li H, Wang Y, Lv X, Zhao F. Comparative Whey Proteome Profiling of Donkey Milk With Human and Cow Milk.. Front Nutr 2022;9:911454.
- Hachana Y, Nasraoui C, Frija I, Fortina R. Arabian mare's milk characterisation and clotting ability.. J Food Sci Technol 2022 May;59(5):1840-1846.
- Polak-Berecka M, Michalak-Tomczyk M, Skrzypczak K, Michalak K, Rachwał K, Waśko A. Potential Biological Activities of Peptides Generated during Casein Proteolysis by Curly Kale (Brassica oleracea L. var. sabellica L.) Leaf Extract: An In Silico Preliminary Study.. Foods 2021 Nov 21;10(11).
- Kocic H, Langerholc T, Kostic M, Stojanovic S, Najman S, Krstic M, Nesic I, Godic A, Wollina U. The Regenerative Potential of Donkey and Human Milk on the Redox-Sensitive and Proliferative Signaling Pathways of Skin Fibroblasts.. Oxid Med Cell Longev 2020;2020:5618127.
- Rodzik A, Pomastowski P, Railean-Plugaru V, Sprynskyy M, Buszewski B. The Study of Zinc Ions Binding to α(S1)-, β- and κ-Casein.. Int J Mol Sci 2020 Oct 30;21(21).
- Faccia M, Gambacorta G, Martemucci G, Difonzo G, D'Alessandro AG. Chemical-Sensory Traits of Fresh Cheese Made by Enzymatic Coagulation of Donkey Milk.. Foods 2019 Dec 23;9(1).
- Polidori P, Vincenzetti S. Use of Donkey Milk in Children with Cow's Milk Protein Allergy.. Foods 2013 May 6;2(2):151-159.
- Moosavi-Movahedi Z, Gharibi H, Hadi-Alijanvand H, Akbarzadeh M, Esmaili M, Atri MS, Sefidbakht Y, Bohlooli M, Nazari K, Javadian S, Hong J, Saboury AA, Sheibani N, Moosavi-Movahedi AA. Caseoperoxidase, mixed β-casein-SDS-hemin-imidazole complex: a nano artificial enzyme.. J Biomol Struct Dyn 2015;33(12):2619-32.
- Vincenzetti S, Polidori P, Salimei E, Mariani PL, Allegri S, Cammertoni N, Quadrini B, Vita A. Purification and identification of alphas1- and beta-caseins from asses milk.. Vet Res Commun 2005 Aug;29 Suppl 2:211-3.
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