Techno-functional properties of clover grass protein and the effect of different process technologies

Badfar, N.; Gregersen Echers, S.; Jacobsen, C.; Yesiltas, B.; Jorgensen, A. K.; Mattsson, T.; Lubeck, P. S.; Mishra, A.; Sancho, A. I.; Bogh, K. L.; Lubeck, M.

doi:10.64898/2026.01.27.701969

bioRxiv · 10.64898/2026.01.27.701969

Techno-functional properties of clover grass protein and the effect of different process technologies

Badfar, N.·Gregersen Echers, S.·Jacobsen, C.·Yesiltas, B.·Jorgensen, A. K.·Mattsson, T.·Lubeck, P. S.·Mishra, A.·Sancho, A. I.·Bogh, K. L.·Lubeck, M.

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Abstract

This study investigated the effects of different downstream processes for protein isolation on the bulk properties and composition of clover grass protein prototypes (CGPs). A clarified clover grass juice, obtained using membrane filtration (MF), underwent precipitation by acid (AP), heat (HP), or acid+heat (AHP), or underwent ultra- and diafiltration to produce a concentrate (DC) as well as subsequent tryptic hydrolysis of DC (DCH). HP had the highest protein content (p<0.05) and was whiter than other CGPs, although it showed lower aqueous solubility. In contrast, DC showed excellent solubility across a broad pH range. CGPs efficiently decreased oil-water interfacial tension (16-13 mN/m) and displayed viscoelastic and solid-like interfacial behavior. CGPs-stabilized emulsions displayed low physical stability with larger droplets despite high absolute {zeta}-potentials. CGPs were rich in RuBisCO (37-47%) but had varying levels of other proteins. Despite significant protein-level differences, overall protein composition of CGPs was comparable, highlighting that protein state governs bulk functionality more than subtle compositional changes. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/701969v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@f37756org.highwire.dtl.DTLVardef@1fb5beorg.highwire.dtl.DTLVardef@1d4efe2org.highwire.dtl.DTLVardef@d11ef8_HPS_FORMAT_FIGEXP M_FIG C_FIG Created with BioRender.com HighlightsO_LIThe effect of different processes on functional properties of CGPs was explored. C_LIO_LIHeat treatment increased protein purity and whiteness at the expense of solubility. C_LIO_LICGPs efficiently reduced O/W interfacial tension but produced unstable emulsions. C_LIO_LICGPs were found rich in RuBisCO (34-47%) using quantitative proteomics. C_LIO_LIProtein state had larger influence on functionality than protein-level composition. C_LI

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Badfar, N., Gregersen Echers, S., Jacobsen, C., Yesiltas, B., Jorgensen, A. K., Mattsson, T., Lubeck, P. S., Mishra, A., Sancho, A. I., Bogh, K. L., Lubeck, M.. 2026-01-29. Techno-functional properties of clover grass protein and the effect of different process technologies. https://doi.org/10.64898/2026.01.27.701969

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