The in situ fortification of vitamin B12 (VB12) in foods through fermentation is an effective strategy to address the deficiency of this micronutrient, and precise monitoring of VB12 production is crucial for developing VB12-fortified functional foods. Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is advantageous for analyzing trace substances in food due to its high sensitivity. In the present study, an LC-MS/MS method capable of rapidly and accurately quantifying three active forms of VB12, namely adenosylcobalamin (AdoCbl), methylcobalamin (MeCbl), hydroxocobalamin (OHCbl), in 8 min were developed. Meanwhile, the quantitative result of this method is not affected by pseudo-VB12 because the selected ion channels include fragments of active VB12. Maintaining light-shielding during extraction and purification is essential, as light exposure during the process can decrease the content of detected VB12 by about 30%. At last, the developed method was applied for the determination of VB12 in fermented rice bran and the cell mass of Propionibacterium freudenreichii. The results showed that AdoCbl was the predominant form of VB12 during fermentation, and the addition of cobalt did not influence the proportions of the three VB12 types. The present study reported a rapid and accurate method for the simultaneous determination of three active forms of VB12, which can effectively support the development of foods with VB12 fortification.

To improve the high-value application of millet bran, a water-soluble polysaccharide was extracted from fermented millet bran (FMBP) by using Bacillus natto fermentation. A neutral polysaccharide, FMBP-1, was separated and purified from FMBP using an anion exchange column. Its structure and antioxidant activity in vitro were characterized and determined. The molecular weight of FMBP-1 was 1.154 × 10⁴ Da, and its molecular weight distribution was relatively uniform. The monosaccharide composition, FT-IR, methylation, and NMR results indicated that FMBP-1 was only composed of glucose and was an α-(1→4)-D-glucan that branched at O-6 with a terminal 1-linked α-D-Glcp as a side chain. In addition, the antioxidant assays indicated that FMBP-1 possessed certain capacities for scavenging free radicals and reducing power, and this was in a concentration-dependent manner. This research will provide fundamental data regarding the structure–activity relationship of millet bran polysaccharides and provide a theoretical foundation for the high-value utilization of millet bran within the food and pharmaceutical industries.

The aqueous extract of Lycium ruthenicum Murray (LRE) could attenuate neuroinflammation in mice induced by a high-fat and high-fructose diet (HFFD). Moreover, LRE could adjust bile acid (BA) metabolism and the gut microbiota. Behavioral tests revealed that LRE prevented HFFD-induced cognitive deficits. The treatment of LRE resulted in a decreased expression of inflammation-related mRNA of TNF-α, IL-6, and IL-1β in the cerebral cortex and hippocampus. Furthermore, LRE ameliorated gut microbiota disorder caused by HFFD by markedly elevating the relative abundances of Streptococcus and probiotics such as Lactococcus. Concurrently, it reduced the relative abundances of Helicobacter and Clostridium_XIVa. The levels of tauroursodeoxycholic acid, known for its neuroprotective property, and taurocholic acid, recognized as an anti-inflammatory agent, were significantly enhanced in the hippocampus and cerebral cortex due to the treatment with LRE. In a word, LRE might have the potential to alleviate HFFD-induced cognitive dysfunction by modulating intestinal microbiota and promoting the synthesis of neuroprotective BAs.

Background: Soy allergy is an important nutritional and health issue that needs to be addressed. Lactic acid bacteria (LAB) fermentation is an effective approach to reduce soy protein allergy. Polysaccharides are commonly used in LAB-fermented products to enhance their textural properties. This study proposes a new strategy for developing hypoallergenic soy protein products. Methods: We prepared a soy protein isolate (SPI) through fermentation with LAB (FSPI) and with five types of polysaccharides supplementation, namely polydextrose (PDX), inulin (IN), long-chain inulin (LCIN), soluble soy polysaccharides (SSPS), and β-glucan (BG). The texture and microstructure of different samples were analyzed. Antigenicity and IgE-binding capacity were determined using ELISA. Finally, peptide sequencing was used to identify the degradation degree and frequency of allergenic epitopes. Results: Samples with added PDX (F-PDX) and IN (F-IN) exhibited lower hardness; smaller, irregular pores; and a honeycomb structure, whereas samples with SSPS (F-SSPS) and BG (F-BG) had higher hardness; large, regular pores; and strong sheet structures. The antigenicity and IgE-binding capacity of F-PDX and F-IN were lower both before and after 120 min of in vitro dynamic gastrointestinal digestion. The peptidomics results indicated that F-PDX and F-IN primarily facilitated the degradation of the glycinin G1 and G2 subunits, β-conglycinin α, and the β subunit. Moreover, it increased the frequency of destruction of allergenic epitopes, and further promoted the degradation of epitopes in the external α-helix structures of glycinin and β-conglycinin compared to FSPI. Conclusions: The addition of polysaccharides had a significant impact on the structure and allergenicity of the soy protein gel, especially PDX and IN.