Stability and transepithelial transport of oligopeptide(KRQKYD)with hepatocyte-protective activity from Jinhua ham in human intestinal Caco-2 monolayer cells

The study evaluated the stability of an oligopeptide(Lys-Arg-Gln-Lys-Tyr-Asp,KRQKYD)and its transport mechanism by simulating gastrointestinal digestion and a model of human intestinal Caco-2 monolayer cells in vitro.In this study,the effects of environmental factors(temperature,pH and NaCl concentration)and simulated gastrointestinal digestion on the stability of KRQKYD were evaluated by indicators of the levels of alanine transaminase(ALT),aspartate transaminase(AST)and malondialdehyde(MDA)in an alcoholinduced hepatocyte injury model.The results showed that KRQKYD still maintained satisfactory hepatocyteprotective activity after treatment with different temperatures(20-80℃),pH(3.0-9.0),NaCl concentration(1%-7%)and simulated gastrointestinal digestion,which indicated that KRQKYD showed good stability to environmental factors and simulated gastrointestinal digestion.Furthermore,the intact KRQKYD could be absorbed in a model of Caco-2 monolayer cells with a P_(app)value of(9.70±0.53)×10^(-7)cm/s.Pretreatment with an energy inhibitor(sodium azide),a competitive peptide transporter inhibitor(Gly-Pro)and a transcytosis inhibitor wortmannin did not decrease the level of transepithelial KRQKYD transport,indicating that the transport mechanism of KRQKYD was not associated with energy dependent,vector mediated and endocytosis.The tight junction disruptor cytochalasin D significantly increased the level of transepithelial KRQKYD transport(P<0.05),suggesting that intact KRQKYD was absorbed by paracellular transport.

Influence of soy protein isolate on the gel properties of walnut protein isolate-κ-carrageenan treated with NaCl

The demand for plant protein is increasing significantly due to the shortage of protein resources.Walnut protein,the main by-product of preparing walnut oil,has limited application in the food industry due to its poor solubility.It was found that the soy protein isolate(SPI)concentration had significant effects on the gel properties of the walnut protein isolate(WNPI)-κ-Carrageenan(KC)composite system treated with 15 mmol/L NaCl.The results showed that the gel strength of the composite system increased first and then decreased with the increased concentration of SPI from 0 to 2.5%.The best rheological properties,texture properties,water holding capacity((92.03±1.05)%),swelling ratio((2.04±0.19)%),freeze-thaw stability and thermal stability(85.53°C)of the composite gel were found at an SPI concentration of 1%.In the meantime,the secondary structure of protein had the least α-helix content of 10.17% and the highest β-sheet content of 39.64%,the fluorescence intensity and free sulfhydryl content reached the highest value.1% SPI could also act as a filler for WNPI to enhance the intermolecular forces such as hydrophobic interaction between the two substances,thus forming a stable gel network structure.This study can provide technical support for improving the gel properties of walnut protein and producing new plant protein gel products.