Skip to main content
Log in

Polymorphism at donkey β-lactoglobulin II locus: identification and characterization of a new genetic variant with a very low expression

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

In the last years, donkey milk had evidenced a renewed interest as a potential functional food and a breast milk substitute. In this light, the study of the protein composition assumes an important role. In particular, β-lactoglobulin (β-LG), which is considered as one of the main allergenic milk protein, in donkey species consists of two molecular forms, namely β-LG I and β-LG II. In the present research, a genetic analysis coupled with a proteomic approach showed the presence of a new allele, here named F, which is apparently associated with a null or a severely reduced expression of β-LG II protein. The new β-LG II F genetic variant shows a theoretical average mass (Mav) of 18,310.64 Da, a value practically corresponding with that of the variant D (∆mass < 0.07 Da), but differs from β-LG II D for two amino acid substitutions: Thr100 (variant F) → Ala100 (variant D) and Thr118 (variant F) → Met118 (variant D). Proteomic investigation of the whey protein fraction of an individual milk sample, homozygous FF at β-LG II locus, allowed to identify, as very minor component, the new β-LG II F genetic variant. By MS/MS analysis of enzymatic digests, the sequence of the β-LG II F was characterized, and the predicted genomic data confirmed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ballabio C, Chessa S, Rignanese D, Gigliotti C, Pagnacco G, Terracciano L, Fiocchi A, Restani P, Caroli M (2011) Goat milk allergenicity as a function of αS1-casein genetic polymorphism. J Dairy Sci 94:998–1004. https://doi.org/10.3168/jds.2010-3545

    Article  CAS  PubMed  Google Scholar 

  • Bertino E, Gastaldi D, Monti G, Baro C, Fortunato D, Perono Garoffo L, Coscia A, Fabris C, Mussap M, Conti A (2010) Detailed proteomic analysis on DM: insight into its hypoallergenicity. Front Biosci 2:526–536

    Article  Google Scholar 

  • Brumini D, Criscione A, Bordonaro S, Vegarud GE, Marletta D (2016) Whey proteins and their antimicrobial properties in donkey milk: a brief review. Dairy Sci Technol 96:1–14. https://doi.org/10.1007/s13594-015-0246-1

    Article  CAS  Google Scholar 

  • Chianese L, De Simone C, Ferranti P, Mauriello R, Costanzo A, Quarto M, Garro G, Picariello G, Mamone G, Ramunno L (2013) Occurrence of qualitative and quantitative polymorphism at donkey betalactoglobulin II locus. Food Res Int 54:1273–1279. https://doi.org/10.1016/j.foodres.2012.11.005

    Article  CAS  Google Scholar 

  • Claeys WL, Verraes C, Cardoen S, De Block J, Huyghebaert A, Raes K, Dewettinck K, Herman L (2014) Consumption of raw or heated milk from different species: an evaluation of the nutritional and potential health benefits. Food Control 42:188–201

    Article  Google Scholar 

  • Conti A, Godovac-Zimmermann J, Liberatori J, Braunitzer G (1984) The primary structure of monomeric beta-lactoglobulin I from horse colostrum (Equus caballus, Perissodactyla). Hoppe Seylers Z Physiol Chem 365:1393–1401

    Article  CAS  PubMed  Google Scholar 

  • Criscione A, Cunsolo V, Bordonaro S, Guastella AM, Saletti R, Zuccaro A, D’Urso G, Marletta D (2009) Donkeys’ milk protein fraction investigated by electrophoretic methods and mass spectrometric analysis. Int Dairy J 19:190–197. https://doi.org/10.1016/j.idairyj.2008.10.015

    Article  CAS  Google Scholar 

  • Cunsolo V, Costa A, Saletti R, Muccilli V, Foti S (2007) Detection and sequence determination of a new variant beta-lactoglobulin II from donkey. Rapid Commun Mass Spectrom 21:1438–1446. https://doi.org/10.1002/rcm.2978

    Article  CAS  PubMed  Google Scholar 

  • Cunsolo V, Cairone E, Fontanini D, Criscione A, Muccilli V, Saletti R, Foti S (2009a) Sequence determination of alpha(s1)-casein isoforms from donkey by mass spectrometric methods. J Mass Spectrom 44:1742–1753. https://doi.org/10.1002/jms.1683

    CAS  PubMed  Google Scholar 

  • Cunsolo V, Cairone E, Saletti R, Muccilli V, Foti S (2009b) Sequence and phosphorylation level determination of two donkey beta-caseins by mass spectrometry. Rapid Commun Mass Spectrom 23:1907–1916. https://doi.org/10.1002/rcm.4087

    Article  CAS  PubMed  Google Scholar 

  • Cunsolo V, Muccilli V, Saletti R, Foti S (2013) MALDI-TOF mass spectrometry for the monitoring of she-donkey’s milk contamination or adulteration. J Mass Spectrom 48:148–153

    Article  CAS  PubMed  Google Scholar 

  • Cunsolo V, Muccilli V, Saletti R, Foti S (2014) Mass spectrometry in food proteomics: a tutorial. J Mass Spectrom 49:768–784. https://doi.org/10.1002/jms.3374

    Article  CAS  PubMed  Google Scholar 

  • Cunsolo V, Saletti R, Muccilli V, Gallina S, Di Francesco A, Foti S (2017) Proteins and bioactive peptides from donkey milk: the molecular basis for its reduced allergenic properties. Food Res Int. https://doi.org/10.1016/j.foodres.2017.07.002

    PubMed  Google Scholar 

  • European Commission. 2010. Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. In: Official Journal of the European Union. L 276/33. 20.10.2010

  • Godovac-Zimmermann J, Conti A, Liberatori J, Braunitzer G (1985) The amino-acid sequence of beta-lactoglobulin II from horse colostrum (Equus caballus, Perissodactyla): beta-lactoglobulins are retinol-binding proteins. Biol Chem Hoppe Seyler 366:601–608

    Article  CAS  PubMed  Google Scholar 

  • Godovac-Zimmermann J, Conti A, James L, Napolitano L (1988) Microanalysis of the amino-acid sequence of monomeric beta-lactoglobulin I from donkey (Equus asinus) milk. Biol Chem H-S 369:171–179

    CAS  Google Scholar 

  • Godovac-Zimmermann J, Conti A, Sheil M, Napolitano L (1990) Covalent structure of the minor monomeric β-lactoglobulin II component from donkey milk. Biol Chem H-S 371:871–879

    CAS  Google Scholar 

  • Goraczko AJ (2005) Molecular mass and location of the most abundant peak of the molecular ion isotopomeric cluster. J Mol Model 11:271–277

    Article  CAS  PubMed  Google Scholar 

  • Herrouin M, Mollé D, Faunquant J, Ballestra F, Maubois JL, Léonil J (2000) New variants identified in donkey’s milk whey proteins. J Protein Chem 19:105–115

    Article  CAS  PubMed  Google Scholar 

  • Horn DM, Zubarev RA, McLafferty FW (2000) Automated reduction and interpretation of high resolution electrospray mass spectra of large molecules. J Am Soc Mass Spectrom 11:320–332

    Article  CAS  PubMed  Google Scholar 

  • Malacarne M, Criscione A, Franceschi P, Tumino S, Bordonaro S, Di Frangia F, Marletta D, Summer A (2017) Distribution of Ca, P and Mg and casein micelle mineralisation in donkey milk from the second to ninth month of lactation. Int Dairy J 66:1–5

    Article  CAS  Google Scholar 

  • Mansueto P, Iacono G, Taormina G, Seidita A, D’Alcamo A, Adragna F, Randazzo G, Carta Rini G, Carroccio A (2013) Ass’s milk in allergy to cow’s milk protein: a review. Acta Med Mediterr 29:153–160

    Google Scholar 

  • Marletta D, Criscione A, Bordonaro S, Guastella AM, D’Urso G (2007) Casein polymorphism in goat’s milk. Le Lait 87:491–504

    Article  CAS  Google Scholar 

  • Marletta D, Tidona F, Bordonaro S (2016) Donkey milk proteins: digestibility and nutritional significance. In: “Milk Proteins—From Structure to Biological Properties and Health Aspects” pp 199–209. https://doi.org/10.5772/62597

  • McCarthy C, Carrea A, Diambra L (2017) Bicodon bias can determine the role of synonymous SNPs in human diseases. BMC Genom 18:227. https://doi.org/10.1186/s12864-017-3609-6

    Article  Google Scholar 

  • Medhammar E, Wijesinha-Bettoni R, Stadlmayr B, Nilsson E, Charrondiere UR, Burlingame B (2012) Composition of milk from minor dairy animals and buffalo breeds: a biodiversity perspective. J Sci Food Agr 92:445–474. https://doi.org/10.1002/jsfa.4690

    Article  CAS  Google Scholar 

  • Mitra S, Ray SK, Banerjee R (2016) Synonymous codons influencing gene expression in organisms. Rese Rep Biochem 6:57–65

    Article  Google Scholar 

  • Monti G, Viola S, Baro C, Cresi F, Tovo PA, Moro G, Ferrero MP, Conti A, Bertino E (2012) Tolerability of donkey’s milk in 92 highly-problematic cow’s milk allergic children. J Biol Regul Homeost Agents 26:75–82

    CAS  PubMed  Google Scholar 

  • Ochirkhuyag B, Chobert JM, Dalgalarrondo M, Haertlé T (2000) Characterization of mare caseins. Identification of αs1- and αs2-caseins. Lait 80:223–235

    Article  CAS  Google Scholar 

  • Restani P, Ballabio C, Di Lorenzo C (2009) Molecular aspects of milk allergens and their role in clinical events. Anal Bioanal Chem 395:47–56

    Article  CAS  PubMed  Google Scholar 

  • Saletti R, Muccilli V, Cunsolo V, Fontanini D, Capocchi A, Foti S (2012) MS-based characterization of as2-casein isoforms in donkey’s milk. J Mass Spectrom 47:1150–1159. https://doi.org/10.1002/jms.3031

    Article  CAS  PubMed  Google Scholar 

  • Salimei E, Fantuz F (2012) Equid milk for human consumption. Int Dairy J 24:130–142. https://doi.org/10.1016/j.idairyj.2011.11.008

    Article  CAS  Google Scholar 

  • Senko MW, Beu SC, McLafferty FW (1995) Determination of monoisotopic masses and ion populations for large biomolecules from resolved isotopic distributions. J Am Soc Mass Spectrom 6:229–233

    Article  CAS  PubMed  Google Scholar 

  • Skoog B, Wichman A (1986) Calculation of the isoelectric points of polypeptides from the amino acid composition. Trends Anal Chem 5:82–83

    Article  CAS  Google Scholar 

  • Swaisgood HE (1995) Protein and amino acid composition of bovine milk. In: Jensen RC (ed) Handbook of milk composition. Academic Press Inc., San Diego, pp 464–468

    Chapter  Google Scholar 

  • Swar MO (2011) Donkey milk-based formula: a substitute for patients with cow’s milk protein allergy. Sudan J Paediatr 11:21–24

    Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular Evolutionary Genetics Analysis Version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tidona F, Sekse C, Criscione A, Jacobsen M, Bordonaro S, Marletta D, Vegarud GE (2011) Antimicrobial effect of donkeys’ milk digested in vitro with human gastrointestinal enzymes. Int Dairy J 21:158–165. https://doi.org/10.1016/j.idairyj.2010.10.008

    Article  CAS  Google Scholar 

  • Tidona F, Criscione A, Devold TG, Bordonaro S, Marletta D, Vegarud GE (2014) Protein composition and micelle size of donkey milk with different protein patterns: effects on digestibility. Int Dairy J 35:57–62. https://doi.org/10.1016/j.idairyj.2013.10.018

    Article  CAS  Google Scholar 

  • Vincenzetti S, Foghini L, Pucciarelli S, Polzonetti Cammertoni N, Beghelli D, Polidori P (2014) Hypoallergenic properties of donkey’s milk: a preliminary study. Vet Ital 50:99–107. https://doi.org/10.12834/VetIt.219.125.5

    PubMed  Google Scholar 

  • Wal JM (2002) Cow’s milk proteins/allergens. Ann Allergy Asthma Immunol 89:3–10. https://doi.org/10.1016/S1081-1206(10)62115-1

    Article  CAS  PubMed  Google Scholar 

  • Waserman S, Watson W (2011) Food allergy. Allergy Asthma Clin Immunol 7(Suppl 1):57. https://doi.org/10.1186/1710-1492-7-S1-S7

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from PO FERS 2007/13 4.1.2.A, project “Piattaforma regionale di ricerca translazionale per la salute”, CUP B65E12000570008. The authors gratefully acknowledge the Bio-Nanotech Research and Innovation Tower (BRIT; PON project financed by the Italian Ministry for Education, University and Research, MIUR) for the availability of the Orbitrap Fusion mass spectrometer.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vincenzo Cunsolo.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Additional information

Handling Editor: P. Beltran-Alvarez.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 4504 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Criscione, A., Cunsolo, V., Tumino, S. et al. Polymorphism at donkey β-lactoglobulin II locus: identification and characterization of a new genetic variant with a very low expression. Amino Acids 50, 735–746 (2018). https://doi.org/10.1007/s00726-018-2555-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-018-2555-1

Keywords

Navigation