Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity

<b>Background</b>: During the initial steps of green biorefining aimed at protein recovery, endogenous proteins and enzymes, along with, e.g., phytochemical constituents, are decompartmentalized into a green juice. This creates a highly dynamic environment prone to a plethora of reaction...

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Main Authors: Kathrine Danner Aakjær Pedersen, Line Thopholm Andersen, Mads Heiselberg, Camilla Agerskov Brigsted, Freja Lyngs Støvring, Louise Mailund Mikkelsen, Sofie Albrekt Hansen, Christian Enrico Rusbjerg-Weberskov, Mette Lübeck, Simon Gregersen Echers
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Language:English
Published: MDPI AG 2025-02-01
Series:Proteomes
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Online Access:https://www.mdpi.com/2227-7382/13/1/8
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author Kathrine Danner Aakjær Pedersen
Line Thopholm Andersen
Mads Heiselberg
Camilla Agerskov Brigsted
Freja Lyngs Støvring
Louise Mailund Mikkelsen
Sofie Albrekt Hansen
Christian Enrico Rusbjerg-Weberskov
Mette Lübeck
Simon Gregersen Echers
author_facet Kathrine Danner Aakjær Pedersen
Line Thopholm Andersen
Mads Heiselberg
Camilla Agerskov Brigsted
Freja Lyngs Støvring
Louise Mailund Mikkelsen
Sofie Albrekt Hansen
Christian Enrico Rusbjerg-Weberskov
Mette Lübeck
Simon Gregersen Echers
author_sort Kathrine Danner Aakjær Pedersen
collection DOAJ
description <b>Background</b>: During the initial steps of green biorefining aimed at protein recovery, endogenous proteins and enzymes, along with, e.g., phytochemical constituents, are decompartmentalized into a green juice. This creates a highly dynamic environment prone to a plethora of reactions including oxidative protein modification and deterioration. Obtaining a fundamental understanding of the enzymes capable of exerting antioxidant activity <i>ex vivo</i> could help mitigate these reactions for improved product quality. <b>Methods</b>: In this study, we investigated perennial ryegrass (<i>Lolium perenne</i> var. Abosan 1), one of the most widely used turf and forage grasses, as a model system. Using size exclusion chromatography, we fractionated the green juice to investigate <i>in vitro</i> antioxidant properties and coupled this with quantitative bottom-up proteomics, GO-term analysis, and fraction-based enrichment. <b>Results</b>: Our findings revealed that several enzymes, such as superoxide dismutase and peroxiredoxin proteoforms, already known for their involvement in <i>in vivo</i> oxidative protection, are enriched in fractions displaying increased <i>in vitro</i> antioxidant activity, indicating retained activity <i>ex vivo</i>. Moreover, this study provides the most detailed characterization of the <i>L. perenne</i> proteome today and delivers new insights into protein-level partitioning during wet fractionation. <b>Conclusions</b>: Ultimately, this work contributes to a better understanding of the first steps of green biorefining and provides the basis for process optimization.
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spelling doaj-art-6297e52e0b5a43afb63d3393baa8cb342025-08-20T03:44:02ZengMDPI AGProteomes2227-73822025-02-01131810.3390/proteomes13010008Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant ActivityKathrine Danner Aakjær Pedersen0Line Thopholm Andersen1Mads Heiselberg2Camilla Agerskov Brigsted3Freja Lyngs Støvring4Louise Mailund Mikkelsen5Sofie Albrekt Hansen6Christian Enrico Rusbjerg-Weberskov7Mette Lübeck8Simon Gregersen Echers9Department of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, DenmarkDepartment of Chemistry & Bioscience, Aalborg University, Fredrik Bajers Vej 7H, DK-9220 Aalborg, Denmark<b>Background</b>: During the initial steps of green biorefining aimed at protein recovery, endogenous proteins and enzymes, along with, e.g., phytochemical constituents, are decompartmentalized into a green juice. This creates a highly dynamic environment prone to a plethora of reactions including oxidative protein modification and deterioration. Obtaining a fundamental understanding of the enzymes capable of exerting antioxidant activity <i>ex vivo</i> could help mitigate these reactions for improved product quality. <b>Methods</b>: In this study, we investigated perennial ryegrass (<i>Lolium perenne</i> var. Abosan 1), one of the most widely used turf and forage grasses, as a model system. Using size exclusion chromatography, we fractionated the green juice to investigate <i>in vitro</i> antioxidant properties and coupled this with quantitative bottom-up proteomics, GO-term analysis, and fraction-based enrichment. <b>Results</b>: Our findings revealed that several enzymes, such as superoxide dismutase and peroxiredoxin proteoforms, already known for their involvement in <i>in vivo</i> oxidative protection, are enriched in fractions displaying increased <i>in vitro</i> antioxidant activity, indicating retained activity <i>ex vivo</i>. Moreover, this study provides the most detailed characterization of the <i>L. perenne</i> proteome today and delivers new insights into protein-level partitioning during wet fractionation. <b>Conclusions</b>: Ultimately, this work contributes to a better understanding of the first steps of green biorefining and provides the basis for process optimization.https://www.mdpi.com/2227-7382/13/1/8perennial ryegrassgreen biorefiningwet fractionationantioxidant activitysize fractionationbottom-up proteomics
spellingShingle Kathrine Danner Aakjær Pedersen
Line Thopholm Andersen
Mads Heiselberg
Camilla Agerskov Brigsted
Freja Lyngs Støvring
Louise Mailund Mikkelsen
Sofie Albrekt Hansen
Christian Enrico Rusbjerg-Weberskov
Mette Lübeck
Simon Gregersen Echers
Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
Proteomes
perennial ryegrass
green biorefining
wet fractionation
antioxidant activity
size fractionation
bottom-up proteomics
title Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
title_full Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
title_fullStr Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
title_full_unstemmed Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
title_short Identifying Endogenous Proteins of Perennial Ryegrass (<i>Lolium perenne</i>) with <i>Ex Vivo</i> Antioxidant Activity
title_sort identifying endogenous proteins of perennial ryegrass i lolium perenne i with i ex vivo i antioxidant activity
topic perennial ryegrass
green biorefining
wet fractionation
antioxidant activity
size fractionation
bottom-up proteomics
url https://www.mdpi.com/2227-7382/13/1/8
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