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. 2024 Nov 20:25:102025.
doi: 10.1016/j.fochx.2024.102025. eCollection 2025 Jan.

Identification and comparison of N-glycome profiles from common dietary protein sources

Matthew Bolino  1 , İzzet Avcı  2 , Hacı Mehmet Kayili  3 , Hatice Duman  4 , Bekir Salih  2 , Sercan Karav  4 , Steven A Frese  1   5
Affiliations

Identification and comparison of N-glycome profiles from common dietary protein sources

Matthew Bolino et al. Food Chem X. .

Abstract

The N-glycomes of bovine whey, egg white, pea, and soy protein isolates are described here. N-glycans from four protein isolates were analyzed by HILIC high performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (HILIC-FLD-QTOF-MS/MS). In total, 33 N-glycans from bovine whey and egg white and 10 N-glycans from soy and pea glycoproteins were identified. The type of N-glycans per glycoprotein source were attributable to differences in biosynthetic glycosylation pathways. Animal glycoprotein sources favored a combination of complex and hybrid glycan configurations, while the plant proteins were dominated by oligomannosidic N-glycans. Bovine whey glycoprotein isolate contained the most diverse N-glycans by monosaccharide composition as well as structure, while plant sources such as pea and soy glycoprotein isolates contained an overlap of oligomannosidic N-glycans. The results suggest N-glycan structure and composition is dependent on the host organism which are driven by the differences in N-glycan biosynthetic pathways.

Keywords: Glycan; Mass spectrometry; Microbiome; N-glycan; N-glycome; Protein.

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Conflict of interest statement

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Steven Frese reports financial support was provided by the United States Department of Agriculture's National Institute of Food and Agriculture. The other authors have no known competing financial interests have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
The most abundant N-glycan structures across protein sources, as determined by mass spectrometry.
Fig. 2
Fig. 2
Distinct N-glycan structures from bovine whey glycoprotein and their relative abundance determined by peak areas from the FLD chromatogram. HILIC-HPLC with a fluorescence detector paired with QTOF-MS/MS produced chromatograms then determined structures and abundance for procainamide-labeled N-glycans from bovine whey protein. A total of 22 peaks were identified corresponding to 33 distinct N-glycan structures. N-glycan structures for low abundant peaks not shown.
Fig. 3
Fig. 3
Distinct N-glycan structures from egg white glycoprotein and their relative abundance determined by peak areas from the FLD chromatogram. HILIC-HPLC with a fluorescence detector paired with QTOF-MS/MS produced chromatograms then determined structures and abundance for procainamide-labeled N-glycans from egg white protein. A total of 30 peaks were identified corresponding to 33 distinct N-glycan structures. N-glycan structures for low abundant peaks not shown.
Fig. 4
Fig. 4
Distinct N-glycan structures from soy glycoprotein and their relative abundance determined by peak areas from the FLD chromatogram. HILIC-HPLC with a fluorescence detector paired with QTOF-MS/MS produced chromatograms and determined structures and abundance for procainamide-labeled N-glycans from soy protein. A total of 10 peaks were identified corresponding to 10 distinct N-glycan structures. N-glycan structures for low abundant peaks not shown.
Fig. 5
Fig. 5
Distinct N-glycan structures from pea glycoprotein and their relative abundance determined by peak areas from the FLD chromatogram. HILIC-HPLC with a fluorescence detector paired with QTOF-MS/MS produced chromatograms and determined structures and abundance for procainamide-labeled N-glycans from pea protein. A total of 10 peaks were identified corresponding to 10 distinct N-glycan structures. N-glycan structures for low abundant peaks not shown.
Fig. 6
Fig. 6
Some N-glycan structures are unique, and others are shared between glycoproteins and glycoprotein sources as determined by HPLC-HILIC-FLD-QTOF-MS/MS. (A) Comparison of N-glycan structures between all glycoprotein sources determined by HPLC-QTOF-MS/MS. (B) Comparison of N-glycan structures between animal and plant sources.
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