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Milk protein based cryogel monoliths as novel encapsulant of probiotic bacteria. Part II: L. rhamnosus GG storage stability and bioactivity under in vitro digestion

ABSTRACT: The aim of the study was to assess the ability of milk protein-based cryogel monoliths to preserve the biological role of Lacticaseibacillus rhamnosus GG (LGG) under cryogenic processing, temperature and water activity controlled static storage and in vitro gastrointestinal transit conditions. Moreover, the matrix disintegration, digestibility, and release of bioactive peptides in the simulated gastrointestinal fluids was determined. The cryogelation process did not confer any significant cellular injuries (losses < 0.5 logCFU g-1) due to the achievement of a vitreous state under the implemented cryogenic conditions. Among the tested storage conditions i.e., temperature (5, 20 and 37 °C) was the most influential parameter of the LGG inactivation kinetics (kLGG = 0.002-0.008 to 0.25-0.56 day-1) followed by the protein composition (i.e., the presence of sodium caseinate alleviated the lethality of LGG). Cryogels stored at aw ≤ 0.33 (found in the vicinity or below Tg) allowed good LGG survivability (kLGG 0.033-0.045 day-1) compared to their rubbery analogues (i.e. aw > 0.33). All cryogels exerted a very limited disintegration during the oral and gastric processing steps helping the LGG cells to overcome the harsh gastric stressors. A complete disintegration of the cryogels was achieved during the intestinal step favouring the release of LGG cells and the advancement of the cryogel matrix digestibility (> 64%). The viable counts of LGG in the intestinal phases remained satisfactorily high (> 8.2 logCFU g-1). Peptidomic profiling of the gastric and intestinal phases confirmed the release of ACE and DPP-IV inhibitory, antimicrobial and antioxidant peptides.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Lactobacillus Rhamnosus Gg

SUBMITTER: Céline LECLERCQ

LAB HEAD: Céline LECLERCQ

PROVIDER: PXD040551 | Pride | 2023-08-18

REPOSITORIES: Pride

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Project description:Germination offers advantages to improve legume protein digestibility as it disintegrates seed structure and hydrolyzes proteins and anti-nutrients. Seed permeability (related to polyphenol content of seed coats) is an important factor affecting the duration of seed germination and its impact in protein digestibility and bioactivity. The objective was to compare the effect of seed germination on protease activity, structure and proteolysis of four selected legumes with contrasting seed coat polyphenol profiles (gray zero tannin [GZL], beluga [BL] and dehulled red [DL] lentils; and zero tannin/low vicine-convicine fava bean [ZF]). Protein hydrolysis was characterized during germination and digestion with respect to proteins, peptides and free amino acids (FAA). In vitro antihypertensive and antioxidant activities of digests were investigated, and peptidomic characterization (HPLC-MS/MS) and identification of bioactive fragments in intestinal digests were performed. Regardless of seed type, germination increased protease activity and reduced levels of phytic acid, trypsin inhibitors and tannins (only in BL). Significant proteolysis of the 7S and 11S globulins and concomitant increase peptides and FAA was observed in all sprouted legumes. Digestion kinetics in sprouts revealed a faster generation of FAA and peptides than in dry seeds, with changes more evident for DL associated to faster imbibition, germination and sprout growth. In contrast, BL sprouts showed the lowest protein digestibility, likely due to lower protease activity levels, seed structure disintegration and higher anti-nutrient levels in comparison to GZL, DL and ZF. Moreover, digestion of sprouts resulted in a higher number of resistant peptides in DL and ZF that matched with previously reported bioactive sequences, suggesting a promising health potential of legume sprouts that was confirmed in vitro. Results suggested that the germination process improved protein digestibility and health promoting potential of lentil and fava bean proteins although these changes were more evident in DL due to its rapid imbibition, faster germination and sprout development. This study will provide important information for either plant breeders to develop legume varieties with permeable seed coats or food producers that could use dehulled seeds for efficient production of sprouts as sustainable food sources of plant proteins with improved nutritional and healthy properties.

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Milk protein based cryogel monoliths as novel encapsulant of probiotic bacteria. Part II: L. rhamnosus GG storage stability and bioactivity under in vitro digestion

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