Abstract

Several factors influence the composition of milk. Among these, genetic variation within and between cattle breeds influences milk protein composition, protein heterogeneity, and their posttranslational modifications. Such variations may further influence technological properties, which are of importance for the utilization of milk into dairy products. Furthermore, these potential variations may also facilitate the production of differentiated products (e.g., related to specific breeds or specific genetic variants). The objective of this study was to investigate the genetic variation and relative protein composition of the major proteins in milk from 6 native Norwegian dairy breeds representing heterogeneity in geographical origin, using the modern Norwegian breed, Norwegian Red, as reference. In total, milk samples from 144 individual cows were collected and subjected to liquid chromatography-electrospray ionization/mass spectrometry-based proteomics for identification of genetic and posttranslational modification isoforms of the 4 caseins (αS1-CN, αS2-CN, β-CN, κ-CN) and the 2 most abundant whey proteins (α-lactalbumin and β-lactoglobulin). Relative quantification of these proteins and their major isoforms, including phosphorylations of αS1-CN and glycosylation of κ-CN, were determined based on UV absorbance. The presence and frequency of genetic variants of the breeds were found to be very diverse and it was possible to identify rare variants of the CN, which, to our knowledge, have not been identified in these breeds before. Thus, αS1-CN variant D was identified in low frequency in 3 of the 6 native Norwegian breeds. In general, αS1-CN was found to be quite diverse between the native breeds, and the even less frequent A and C variants were furthermore detected in 1 and 5 of the native breeds, respectively. The αS1-CN variant C was also identified in samples from the Norwegian Red cattle. The variant E of κ-CN was identified in 2 of the native Norwegian breeds. Another interesting finding was the identification of αS2-CN variant D, which was found in relatively high frequencies in the native breeds. Diversity in more common protein genetic variants were furthermore observed in the protein profiles of the native breeds compared with milk from the high-yielding Norwegian Reds, probably reflecting the more diverse genetic background between the native breeds.

Highlights

  • Conservation of native dairy breeds is important to minimize erosion of genetic diversity

  • The sample set contained 144 milk samples from individual cows collected from 30 different farms in Norway analyzed as individual skim milk samples by liquid chromatography (LC)-electrospray ionization (ESI)/MS Single Q

  • This study identified variants of αS1-CN and αS2-CN that have not previously been identified in the native Norwegian breeds included in this study

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Summary

Introduction

Conservation of native dairy breeds is important to minimize erosion of genetic diversity. The Nordic countries have committed to conserve these animal genetic resources, where many of the native breeds have been classified as endangered (Kierkegaard et al, 2020). Compared with a high number of studies focusing on genetic diversity within and among these breeds, only a limited number of studies have been dedicated to provide phenotypic characterization and even fewer have studied milk compositional traits (Kierkegaard et al, 2020), which are important characteristics in relation to eventual exploitation that could contribute to their preservation. The major proteins in bovine milk consist of the 4 caseins, αS1-CN, αS2-CN, β-CN, and κ-CN, together with the 2 most abundant whey proteins, α-LA and β-LG. The genes encoding the 6 major proteins are polymorphic to different degrees, and more than 47 different milk protein variants have been identified within the Bos genus (Caroli et al, 2009).

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