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Proteomic and peptidomic profiling of spirulina-fortified probiotic powder formulations during in vitro digestion

ABSTRACT: This study reports on the influence of lactic acid fermentation on the proteomic and peptidomic profiles of spirulina protein isolate (SPI)-fortified, freeze-dried powders containing living Lacticaseibacillus rhamnosus GG (LGG) cells during in vitro digestion. For comparison, powders fortified with whey protein isolate (WPI) and pea protein isolate (PPI) were also evaluated. Prior to freeze-drying, the powder precursors were either non-treated or fermented. Capillary SDS–PAGE electropherograms revealed a mild proteolytic effect due to fermentation. C-phycocyanin (SPI) and β-lactoglobulin (WPI) showed the highest resistance to pepsinolysis. All samples were responsive to pancreases, with fermented WPI showing the lowest responsiveness. Fermentation enhanced the degree of hydrolysis (DH) in gastric chymes, whereas in intestinal chymes, DH followed the order SPI > PPI > WPI, with fermentation showing no significant impact. A total of 6, 11, and 52 potential bioactive peptide sequences, associated with various beneficial activities, were identified in the SPI, PPI, and WPI digesta, respectively. The highest amino acid bioaccessibilities were observed for cysteine and methionine in SPI, isoleucine and arginine in PPI, and glycine in WPI. In conclusion, fortifying probiotic formulations with protein isolates offers secondary health benefits, stemming from the release of bioactive peptides and bioaccessible essential amino acids.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Lactobacillus Rhamnosus Gg Arthrospira Platensis Pisum Sativum (garden Pea)

TISSUE(S): Plant Cell

SUBMITTER: Céline LECLERCQ

LAB HEAD: Céline LECLERCQ

PROVIDER: PXD052301 | Pride | 2025-02-05

REPOSITORIES: pride

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Project description:Analysis of non-differentiated Caco-2 intestinal epithelial cell line treated with polydextrose fermentation metabolites fermented for 48 hours in 4-stage in vitro colon simulator, in which the conditions mimic the human proximal, ascending, transverse and distal colon in sequence , as well as with medium, 100 mM NaCl and 5 mM butyrate. Polydextrose, a soluble fiber fermented in colon, was fermented with the in vitro colon simulator in three amounts of 0%, 1% and 2%. Results provide insight into the mechanisms underlying colon cancer cells and a comparison of a complex fiber metabolome to 5 mM butyrate and 100 mM NaCl. Furthermore, the results give insight of dosage effect of increasing the concentration of fiber. High level of dietary fiber has been epidemiologically linked to protection against the risk for developing colon cancer. The mechanisms of this protection are not clear. Fermentation of dietary fiber in the colon results in production of for example butyrate that has drawn attention as a chemopreventive agent. Polydextrose, a soluble fiber that is only partially fermented in colon, was fermented in an in vitro colon simulator, in which the conditions mimic the human proximal, ascending, transverse and distal colon in sequence. The subsequent fermentation metabolome were applied on colon cancer cells, and the gene expression changes studied. Polydextrose fermentation down-regulated classes linked with cell cycle, and affected number of metabolically active cells. Further, up-regulated effects on classes linked with apoptosis implicate that polydextrose fermentation plays a role in induction of apoptosis in colon cancer cells. The up-regulated genes involved also key regulators of lipid metabolism, such as PPARg and PGC-1α. These results offer hypotheses for the mechanisms of two health benefits linked with consumption of dietary fiber, reducing risk of development of colon cancer, and dyslipidemia. Non-differentiated Caco-2 cells were treated with polydextrose fermentation metabolites from the vessels representing different parts of the colon, or with 100 mM NaCl or with 5 mM butyrate for 24 hours. For polydextrose fermentation three concentrations of polydextrose were used: 0%, 1% and 2% for a simulation that lasted for 48 hours. Polydextrose fermentation samples from total of 12 vessels, as well as from medium sample, 5 mM butyrate and 100 mM NaCl were analysed as single replica.

			Action	DRS
	F180668.dat	Other
	F180668.mgf	Mgf
	F180668.mzid.gz	Mzid
	F180669.dat	Other
	F180669.mgf	Mgf

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Proteomic and peptidomic profiling of spirulina-fortified probiotic powder formulations during in vitro digestion

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