| Resveratrol is a polyphenolic compound widely found in plants such as grapes,peanuts and knotweed.It has good physiological functions such as antioxidant,immune regulation,antitumor,anti-inflammatory and cardiovascular protection,and it is also widely used in health foods,pharmaceuticals and other fields.However,resveratrol has the disadvantages of low water solubility and poor stability.At the same time,because it is rapidly metabolized and excreted in the intestine and liver,its bioavailability is very low,which seriously affects the play of physiological activities.In this paper,the Scomberomorus niphonius peptides were prepared from the Scomberomorus niphonius,which can significantly improve the digestion stability and bioavailability of resveratrol,thereby significantly improving its physiological activity.The hydrolysis process of Scomberomorus niphonius was optimized,and the effects of different proteases,ratio of material to liquid,amount of enzyme added,time of hydrolysis,and p H of hydrolysis on the effect of hydrolysis were investigated.The optimal process was determined as follows: alkaline protease was used,the substrate to liquid ratio was 1:10,the ratio of enxyme to substrate ratio was 3000 U/g,the hydrolysis time was 4 h,the hydrolysis p H value was 8,and the hydrolysis temperature was 50 ?,with a protein recovery of 67.6 ± 1.8% and a degree of hydrolysis of 22.5 ± 2.9%.The study compared the inhibitory effect of Scomberomorus niphonius hydrolysis product(SH),resveratrol and Scomberomorus niphonius hydrolysis product-resveratrol complex(SH-R)on B16 cells.It was found that,SH has no toxic effect on B16 cells.At 37 ? and a phenol / peptide ratio of 1:10,SH can increase the inhibition rate of resveratrol on B16 cells by 14.3%(P<0.05).The interaction between SH and resveratrol was characterized by fluorescence spectroscopy.It was found that resveratrol caused obvious fluorescence quenching of SH,and dynamic quenching and static quenching may coexist in the interaction.The number of binding sites between resveratrol and SH was 0.935 at 37 ?,and it mainly shows hydrophobic effect.Using the inhibition rate of the complex on B16 cells as an indicator,SH was further separated and purified.First,using DEAE-cellulose 52 as a filler,the SH was separated and purified by anion-exchange chromatography to obtain the component SHa.Sephadex G-15 was used as the filler and separated and purified by size-exclusion chromatography to obtain the component SHa1.The compound SHa1-R(37 ?,phenol / peptide ratio 1:10)can increase the inhibition rate of resveratrol on B16 cells by 21.7%.Analysis shows that SHa1 can significantly increase the solubility and digestion stability of resveratrol.Using a non-everted rat gut sac model,the effect of SHa1 on the intestinal absorption rate of resveratrol was analyzed.It was found that SHa1 increased the intestinal absorption rate of resveratrol in the duodenum and jejunum by 16.5% and 9.5%.It shows that SHa1 may improve the digestion stability and bioavailability of resveratrol,so that its effect of inhibiting B16 cells is significantly improved.The mechanism of SHa1 to improve the digestive stability and bioavailability of resveratrol was discussed.High-performance liquid chromatography was used to determine the molecular weight distribution of SHa1.It was found that small molecules below 1000 Da accounted for more than 87% of SHa1,and 317-623 Da substances accounted for more than 60% of SHa1.A high-speed amino acid analyzer was used to determine the amino acid composition and content of SHa1,and it was found that it was rich in aspartic acid,lysine,and leucine.Using ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometry to identify the amino acid sequence of SHa1,five peptides were identified: TLR,LQLE,LDDL,LNLT,ALDLH.SHa1 interacts with resveratrol and may have a significant impact on the digestion and absorption characteristics,bioavailability and physiological activity of resveratrol. |
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