Study on the binding behavior and functional properties of soybean protein isolate and ß-carotene.

This study focused on the non-covalent interaction between soybean protein isolate (SPI) and ß-carotene (BC). The conformational changes of SPI with ß-carotene in varying proportions (BC/SPI: 2%, 4%, 6%, 8%, and 10%) were investigated by multi-spectroscopy and molecular docking. Results showed that the quenching mode is static quenching and binding affinity increased with temperature. The stoichiometry was 1:1, indicating there was only one binding site in SPI. The binding was based on entropy and primarily driven by hydrophobic interactions and its binding constant was in the order of 104 L⋅mol-1. The addition of the ß-carotene affected the secondary structure of SPI resulting in an increase in α-Helix and a decrease in random coil and ß-turn content, indicating protein aggregated and hydrophobic interactions occurred. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) verified that no new larger molecular weight substance was formed and no covalent interaction existed. Molecular docking corroborated that electrostatic and hydrophobic interactions were both involved in the formation of complexes, where hydrophobic interaction was the dominant one. Moreover, ß-carotene improved 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, foaming capacity, and emulsifying stability of SPI. These findings provide useful information about the interaction mechanism of SPI and ß-carotene, which contributes to the further development and application of SPI products rich in ß-carotene in the food industry.

Peptidomics comparison of plant-based meat alternatives and processed meat after in vitro digestion.

Plant-based meat alternatives (PMAs) is a new type of food that meets people's health needs, but the lack of awareness of its nutritional properties limits product development and promotion. Here, we compared the similarities and differences of the nutritional properties of PMAs and meat before and after in vitro simulation of gastrointestinal digestion by chemical composition analysis, peptidomics and bioactivity tests. The molecular weights of Plant-based meat alternatives derived peptides (PDPs) as well as meat-derived peptides (MDPs) in the beef and pork groups were mainly concentrated in the low mass range from 800 Da to 1500 Da. The principal component analysis indicated that the composition of MDPs in the beef and pork groups significantly differed from PDPs but overlapped slightly with the chicken group. Also, there were very few common peptides among them. The proportion of high-biological-scoring peptides (33.3%-40%) in PDPs was more than that in MDPs (4.8%-20.8%). PDPs were predicted to have higher antibacterial activity than others. PDPs and MDPs showed a certain antioxidant capacity and angiotensin converting enzyme inhibitory activity (62.2%-92.5%) in vitro. Some peptides weakly inhibited the growth of Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) while promoting the growth of probiotics. This research provides a theoretical basis for in-depth exploration of the nutritional characteristics of PMAs.

Colonization and Gut Flora Modulation of Lactobacillus kefiranofaciens ZW3 in the Intestinal Tract of Mice.

This study evaluated the distribution and colonization of Lactobacillus kefiranofaciens ZW3 and determined its capacity to modulate the gut microbiota in an animal model. Based on (1) fluorescence imaging, (2) flow cytometry, and (3) qPCR, we found that ZW3 successfully adhered to mouse mucous tissue and colonized the mouse ileum. Gut microbiota profiling was performed using high-throughput sequencing. After continuous intubation with ZW3 for 1 week, the proportion of Lachnospiraceae, a family of butyric acid-producing bacteria, increased at day 7 (11.9% at day 0 versus 18.4% at day 7). In addition, Lactobacillaceae showed an increasing trend (4% at day 0 versus 13% at day 7) that was accompanied by an observable decline in the Rikenellaceae family (1.58% at day 7, 0.14% at day 14, and 0.75% at day 21) in the tested mouse. The results demonstrate that ZW3 could successfully adhere to and colonize the mouse gut throughout the course of the experiment. The profiling analysis of the gut microbiota also provided evidence supporting the function of ZW3 in improving the intestinal flora of mice.

Comparative genomics of Lactobacillus kefiranofaciens ZW3 and related members of Lactobacillus. spp reveal adaptations to dairy and gut environments.

It is important for probiotics that are currently utilized in the dairy industry to have clear genetic backgrounds. In this study, the genetic characteristics of Lactobacillus kefiranofaciens ZW3 were studied by undertaking a comparative genomics study, and key genes for adaptation to different environments were investigated and validated in vitro. Evidence for horizontal gene transfer resulting in strong self-defense mechanisms was detected in the ZW3 genome. We identified a series of genes relevant for dairy environments and the intestinal tract, particularly for extracellular polysaccharide (EPS) production. Reverse transcription-qPCR (RT-qPCR) revealed significant increases in the relative expression of pgm, ugp, and uge during the mid-logarithmic phase, whereas the expression of pgi was higher at the beginning of the stationary phase. The enzymes encoded by these four genes concertedly regulated carbon flux, which in turn modulated the production of EPS precursors. Moreover, ZW3 tolerated pH 3.5 and 3% bile salt and retained cell surface hydrophobicity and auto-aggregation. In conclusion, we explored the potential of ZW3 for utilization in both the dairy industry and in probiotic applications. Additionally, we elucidated the regulation of the relevant genes involved in EPS production.

Molecular mechanisms and in vitro antioxidant effects of Lactobacillus plantarum MA2.

Lactobacillus plantarum MA2 was isolated from Chinese traditional Tibetan kefir grains. The antioxidant activities in vitro of this strain were evaluated extensively. The results showed that L. plantarum MA2 can tolerate hydrogen peroxide up to 2.0mM, and its fermentate (fermented supernatant, intact cell and cell-free extract) had strong reducing capacities, lipid peroxidation inhibition capacities, Fe2+-chelating abilities, as well as various free radical scavenging capacities. Additionally, both the fermented supernatant and cell homogenate exhibited glutathione peroxidase activity and superoxide dismutase activity. In order to investigate the antioxidant mechanism of L. plantarum MA2 at the molecular level, eight antioxidant-related genes were identified, and further analyzed. Three groups of genes cat, gshR and npx, were found up-regulated under H2O2 challenge.

In vitro and in vivo evaluation of the probiotic attributes of Lactobacillus kefiranofaciens XL10 isolated from Tibetan kefir grain.

Lactobacillus kefiranofaciens XL10, with a high yield of extracellular polysaccharide (EPS), is isolated from Tibetan kefir grain and benefits the health of human beings and has been considered to exhibit probiotic potential in vitro and in vivo. The probiotic function of the strain was studied extensively, viz., acid and bile salt tolerances, cell surface hydrophobicity and autoaggregation, the modulation of gut microbiota, and the distribution and colonization of XL10 in the mouse intestinal tract after oral administration. XL10 could survive 3-h incubation at pH 3.5 and exhibited cell surface hydrophobicity of ∼79.9% and autoaggregation of ∼27.8%. After continuous oral administration of XL10 for 2 weeks, the Bifidobacteriaceae family increased, accompanied by an observable decline in Proteobacteria phyla in the tested mice. Butyrivibrio and Pseudobutyrivibrio, recognized as butyric acid-producing bacteria, could also be detected at day 7 and day 14, respectively. The most abundant community in the mouse gut had formed by day 14. Additionally, we found that XL10 successfully adhered to the mucous tissue and colonized the ileum of the mice based on fluorescence imaging, flow cytometry, and qPCR. Our results suggested that XL10 has excellent probiotic properties and represents an alternative for exploitation in the development of novel functional foods.

Antioxidative effects in vivo and colonization of Lactobacillus plantarum MA2 in the murine intestinal tract.

Lactobacillus plantarum MA2 was isolated from traditional Chinese Tibet kefir grains, which possess several excellent properties and functions. We previously demonstrated the antioxidant activities of this bacterium in vitro. However, the maintenance and survival of L. plantarum MA2 inside the murine intestinal tract, where it exerts its probiotic properties, and whether its effects are elicited directly on the host remain unknown. Therefore, this study investigated the mechanisms of L. plantarum MA2 in aging mice following D-galactose administration. The levels of malondialdehyde decreased significantly in the L. plantarum MA2 groups after oral ingestion compared to the D-galactose model group, and total antioxidant capacity and glutathione peroxidase and superoxide dismutase activities increased significantly in the serum and liver. We combined fluorescein isothiocyanate labeling and green fluorescent protein expression to dynamically monitor the colonization and distribution of L. plantarum MA2 in the murine intestinal tract. The results indicated that L. plantarum MA2 was detected in the ileum, colon, and feces after single and continuous oral administration at day 21 and was maintained at 10(4)-10(5) CFU/g. These results suggest that L. plantarum MA2 colonizes and survives in the murine intestinal tract to exert its antioxidative effects.

Functional and bioinformatics analysis of an exopolysaccharide-related gene (epsN) from Lactobacillus kefiranofaciens ZW3.

A novel lactic acid bacteria strain Lactobacillus kefiranofaciens ZW3 exhibited the characteristics of high production of exopolysaccharide (EPS). The epsN gene, located in the eps gene cluster of this strain, is associated with EPS biosynthesis. Bioinformatics analysis of this gene was performed. The conserved domain analysis showed that the EpsN protein contained MATE-Wzx-like domains. Then the epsN gene was amplified to construct the recombinant expression vector pMG36e-epsN. The results showed that the EPS yields of the recombinants were significantly improved. By determining the yields of EPS and intracellular polysaccharide, it was considered that epsN gene could play its Wzx flippase role in the EPS biosynthesis. This is the first time to prove the effect of EpsN on L. kefiranofaciens EPS biosynthesis and further prove its functional property.