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