Jack bean (JB), Canavalia ensiformis (L.) DC, is a commonly cultivated legume in Indonesia. It is rich in protein, which can be hydrolyzed, making it potentially a good source of bioactive peptides. Intestinal inflammation is associated with several diseases, and the production of interleukin-8 (IL-8) in intestinal epithelial cells induced by tumor necrosis factor (TNF)-α has an important role in inflammatory reaction. The present study investigated the anti-inflammatory effects of peptides generated from enzymatic hydrolysis of JB protein on human intestinal Caco-2BBe cells. Additionally, in silico approaches were used to identify potential bioactive peptides. JB protein hydrolysate (JBPH) prepared using pepsin and pancreatin reduced the IL-8 expression at protein and mRNA levels in Caco-2BBe cells stimulated with TNF-α. Immunoblot analysis showed that the JBPH reduced the TNF-α-induced phosphorylation of c-Jun-NH(2)-terminal kinase, nuclear factor kappa B (NF-κB), and p38 proteins. Anti-inflammatory activity was observed in the 30% acetonitrile fraction of JBPH separated on a Sep-Pak C18 column. An ultrafiltration method revealed that relatively small peptides (< 3 kDa) had a potent inhibitory effect on the IL-8 production. Purification of the peptides by reversed-phase and anion-exchange high performance chromatography produced three peptide fractions with anti-inflammatory activities. A combination of mass spectrometry analysis and in silico approaches identified the potential anti-inflammatory peptides. Peptides derived from JB protein reduces the TNF-α-induced inflammatory response in Caco-2BBe cells via NF-κB and mitogen-activated protein kinase signaling pathways. Our results may lead to a novel therapeutic approach to promote intestinal health.

The production of inflammatory cytokines such as tumor necrosis factor (TNF)-α by activated macrophage cells plays an important role in the development of intestinal inflammation. The present study investigated the anti-inflammatory effect of the protein hydrolysates prepared from the jack bean (JBPHs), Canavalia ensiformis (L.) DC, using the enzyme Alcalase, in a murine macrophage model, RAW 264.7 cells, which were stimulated by lipopolysaccharides. JBPHs reduced the TNF-α expression at the protein and mRNA levels through the downregulation of cellular signaling pathways involved in nuclear factor kappa B (NF-κB), extracellular signal-regulated kinase (ERK), and p38. A combination of mass spectrometry and in silico approaches identified 10 potential anti-inflammatory peptides in the JBPHs, including LFLLP and DFFL. Interestingly, while LFLLP targeted the NF-κB pathway, DFFL targeted p38 and ERK to suppress the TNF-α production in the RAW 264.7 cells. In addition, LFLLP and DFFL were localized in the cytosol of the cells. These results demonstrated that LFLLP and DFFL were incorporated by RAW 264.7 cells and, at least in part, contributed to the reduction in TNF-α by JBPHs. These peptides isolated from JBPHs may well be utilized as new alternatives to alleviate intestinal inflammation.

Cowpea has high protein content which makes it is potential to be utilized as a protein isolate. However, cowpeas contain some anti-nutritional compounds which can lower the nutritional quality of the cowpeas. It is necessary to reduce the antinutritional contents from the cowpeas.The objective of this study was to investigate the optimum treatments for antinutritional removal and protein extraction to produce protein isolate. Removal of antinutritional was performed by a combination of soaking treatment with the duration of 24, 48, and 72 hours with 5% NaHCO3 solution and boiling for 0, 2.5, and 5 minutes. The protein extraction was conducted by adjusting the pH to various pH at 2, 4, 6, 8, 10, 11, and 12.Soaking of cowpeas seeds for 48 hours with 5% NaHCO3 significantly reduced HCN levels to 0.72 ppm, phytic acid of 9.62 mg / g, and trypsin inhibitor of 7.02 mg / g. The protein concentrate was obtained from protein extraction by using optimum pH at 12 while precipitation of protein by using pH 4 with protein concentration at 70.9%. Sonication with 80%-30 minutes indicated an increase in the water holding capacity, oil holding capacity, emulsion stability, foaming capacity, and stability rather than untreated

Although the mutualistic relationship between the intestinal microbiota and the human host is crucial for maintaining health, the underlying mechanisms of this relationship remain unclear. In the present study, aiming to elucidate the regulatory mechanisms governing the Cyp2c55 expression, which is predominantly observed in colonic tissues, germ‐free, antibiotic‐administered and colitic mice, as well as mouse colonoids, were used as experimental models. RNA sequencing showed comparable decreases in the colonic Cyp2c55 expression in germ‐free and antibiotic‐administered mice, when compared with that in specific pathogen‐free mice. Furthermore, administration of dextran sulfate sodium decreased the Cyp2c55 expression in colitic mice. For these mice, a Pearson correlation analysis also showed a positive correlation between the Cyp2c55 expression and unconjugated bile acids (BAs), including chenodeoxycholic, muricholic, deoxycholic, lithocholic, and ursodeoxycholic acids, as well as taurine (T)‐conjugated secondary BAs, including deoxycholic acid. Moreover, bacterial genera, such as Muribaculaceae and unclassified Lachnospiraceae , also exhibited a positive correlation with these BAs. While administration of an agonist of the pregnane X receptor (PXR) increased the Cyp2c55 expression in mouse colonoids, inflammatory cytokines decreased it. In conclusion, Cyp2c55 was highly expressed in the colonic epithelial cells of mice in a microbiota‐dependent manner. The underlying mechanism seemed to involve a BA‐mediated PXR activation. In addition, the colonic expression of Cyp2c55 was regulated by the inflammatory response. Although the physiological function of Cyp2c55 remains largely unidentified, our findings suggested that Cyp2c55 may play a role in the mutualistic interaction between the intestinal microbiota and the intestinal homeostasis.

Protein binders play a crucial role in determining the quality of emulsified meat products, such as chicken meatballs. Soy protein isolate (SPI) is widely used as a protein binder; however, its production in Indonesia relies predominantly on imported soybeans, underscoring the need to explore alternative protein sources derived from locally available legumes. This study investigated the effects of autoclaving on the antioxidant and techno-functional properties of pigeon pea (Cajanus cajan L. Millsp.) protein hydrolysates (PPHs) and evaluated their potential as a partial substitute for SPI in chicken meatballs. Based on the overall performance across the evaluated antioxidant and techno-functional properties, including DPPH radical-scavenging activity, reducing power, water- and oil-holding capacities, and emulsifying activity and stability, autoclaving at 121 °C for 15 min was selected for subsequent application. Incorporation of PPH into chicken meatballs affected their physical and sensory properties, with increasing PPH proportions resulting in darker color and lower hardness. Despite these changes, the 75:25 PPH:SPI formulation showed overall sensory acceptability comparable to the SPI-only formulation. Overall, autoclaving modified pigeon pea protein and improved several antioxidant and techno-functional properties, supporting its potential as a multifunctional co-protein ingredient for meat-based food formulations and providing a value-added application for an underutilized legume.

Water-soluble soybean fiber (WSSF) exerts health benefits beyond its fermentability, but its direct effects on epithelial-immune interactions remain unclear. We investigated whether WSSF regulates antimicrobial proteins (AMPs) in the small intestine and the underlying mechanisms. C57BL/6J and tuft cell-deficient Pou2f3-knockout mice were fed control or WSSF-supplemented diets, and gene and protein expression and cecal microbiota were analyzed. WSSF upregulated the epithelial AMPs SPRR2A, RELMβ, and ANG4 in the jejunum and ileum. Pharmacological inhibition of group 2 innate lymphoid cells (ILC2) or TRPM5 attenuated AMP induction and reduced IL-25, IL-13, tuft cell markers, and STAT6 phosphorylation. WSSF also failed to induce AMPs or type 2 immune markers in Pou2f3-knockout mice, indicating a requirement for tuft cells. Finally, WSSF altered cecal microbiota composition in a tuft cell-dependent manner. These findings identify WSSF as a soybean-derived fiber that strengthens epithelial innate defense via a TRPM5-tuft cell-ILC2 axis.