Survival and bioactivities of selected probiotic lactobacilli in yogurt fermentation and cold storage: New insights for developing a bi-functional dairy food

In previous work , we demon strated that two probiotic strains, name ly Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, produce fermented milks with potent angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. Here, we tested these strains for the survivability and the release of antihypertensive and antioxidant peptides in yogurt fermentation and cold storage. For these purposes three yogurt batches were compared: one prepared using yogurt starters alone (Lactobacillus delbrueckii subspecies bulgaricus 1932 and Streptococcus thermophilus 99), and the remaining two containing either PRA205 or PRA331 in addition to yogurt starters. Despite the lower viable counts at the fermentation end compared to PRA331, PRA205 overcame PRA331 in survivability during refrigerated storage for 28 days, leading to viable counts (>108 CFU/g) higher than the minimum therapeutic threshold (106 CFU/g). Analyses of in vitro ACE-inhibitory and antioxidant activities of peptide fractions revealed that yogurt supplemented with PRA205 displays higher amounts of antihypertensive and antioxidant peptides than that produced with PRA331 at the end of fermentation and over storage. Two ACE-inhibitory peptides, Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP), were identified and quantified. This study demonstrated that L. casei PRA205 could be used as adjunct culture for producing bi-functional yogurt enriched in bioactive peptides and in viable cells, which bring health benefits to the host as probiotics

Impact of non-starter lactobacilli on release of peptides with angiotensin-converting enzyme inhibitory and antioxidant activities during bovine milk fermentation

This study aimed at evaluating non-starter lactobacilli (NSLAB) isolated from cheeses for their proteolytic activity and capability to produce fermented milk enriched in angiotensin-converting enzyme (ACE)-inhibitory and antioxidant peptides. Preliminarily, 34 NSLAB from Parmigiano Reggiano (PR) and 5 from Pecorino Siciliano cheeses were screened based on their capacity to hydrolyze milk proteins. Two NSLAB strains from PR, Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, showed the most proteolytic phenotype and were positively selected to inoculate sterile cow milk. The fermentation process was monitored by measuring viable cell population, kinetic of acidification, consumption of lactose, and synthesis of lactic acid. Milk fermented with Lb. casei PRA205 exhibited higher radical scavenging (1184.83 ± 40.28 umol/L trolox equivalents) and stronger ACE-inhibitory (IC50 = 54.57 ug/mL) activities than milk fermented with Lb. rhamnosus PRA331 (939.22 ± 82.68 umol/L trolox equivalents; IC50 = 212.38 ug/mL). Similarly, Lb. casei PRA205 showed the highest production of ACE-inhibitory peptides Val-Pro-Pro and Ile-Pro-Pro, which reached concentrations of 32.88 and 7.52 mg/L after 87 and 96 h of milk fermentation, respectively. This evidence supports Lb. casei PRA205, previously demonstrated to possess characteristics compatible with probiotic properties, as a promising functional culture able to promote health benefits in dairy food

Release of the Antihypertensive Tripeptides Valine-Proline-Proline and Isoleucine-Proline-Proline from Bovine Milk Caseins during in Vitro Gastrointestinal Digestion

The aim of this study was to identify and quantify the release of antihypertensive tripeptides valine-proline-proline (VPP) and isoleucine-proline-proline (IPP) during in vitro oro-gastro-intestinal (OGI) digestion of bovine skimmed milk. The experimental approach combined the recently developed harmonized static in vitro digestion (IVD) model and targeted mass spectrometry to monitor peptide generation. We first demonstrated that VPP and IPP are released from bovine milk proteins during in vitro OGI digestion at final concentrations of 354.3 ± 29.8 and 973.8 ± 155.7 μg/L, respectively. In silico analysis of cleavage sites and mass spectrometry revealed that tetrapeptides VPPF, IPPL, and IPPK are precursors of VPP and IPP. The release of other ACE-inhibitory peptides, such as FVAP, VAP, AW, and VY, was demonstrated, and their fate and the time course were investigated. This research underlines the suitability of an IVD system to study the release of short bioactive peptides during OGI transit

Bioactive peptides from vegetable food matrices: Research trends and novel biotechnologies for synthesis and recovery

Currently, the interest for health-promoting functional foods, dietary supplements and pharmaceutical preparations containing bioactive peptides deriving from food proteins, is increasing. Despite the large literature concerning peptides derived from animal proteins, only recently the scientific community investigated the possibility to obtain bioactive peptides from vegetable sources, also discovering novel functional features. In this article, the functional effects of vegetable-derived peptides, including antihypertensive, antioxidant, antitumoral, antiproliferative, hypocholesterolemic, antinflammatory activities, are described. The novel biotechnologies for the release of bioactive peptides from vegetable matrices, including microbial fermentation and the use of microbial enzymes are investigated. Moreover, the modern technologies for their recovery, purification, and analysis, are reviewed and discussed