| In order to increase the added value of wheat and utilize wheat protein efficiently,enzymatic hydrolysis of wheat protein into wheat peptides with certain biological activity and easy absorption is a common treatment method.Due to the low solubility of wheat protein（gluten）,there are problems such as low efficiency,high energy consumption,and difficulty in industrialization with wheat peptide.Therefore,in this study,wheat wet gluten was used as raw material to prepare wheat peptide,and its enzymatic hydrolysis process,parameters were optimized,and its biological activity was analyzed through in vitro activity assay experiments and cell experiments.On this basis,the peptides with relevant activities were screened out with computer-assisted analysis.Finally,the related products were developed according to the biological activity of wheat peptides.The research contents and results were as follows:The wheat peptidase enzymatic hydrolysis process was optimized.Using wheat wet gluten as raw material,amylase-complex protease was used for segmented enzymatic hydrolysis,and the process was further optimized to determine the optimal enzymatic hydrolysis conditions:addingα-amylase（24 U/g starch）to wet gluten at 60℃for 1 h,adjusted the temperature to 55℃,add glucoamylase（250 U/g starch）for enzymatic hydrolysis for 45 min,washed with water to remove reducing sugars produced by enzymatic hydrolysis,and added Alcalase(2.4×10-3AU/g),papain（800 U/g）and bromelain（600 U/g）,enzymatically hydrolyzed with stirring at 50℃for 2 h,and spray dried after high temperature inactivation.And then there was an effective result,the protein（dry basis）content of wheat peptides was 74.6%,and the peptide content reached 61.9%,.The biological activities of wheat peptides were determined and screened.The results of in vitro activity assay showed that wheat peptides had certain antioxidant ability and hypoglycemic activities,among which Dipeptidy peptidase-IV（DPP-IV）had the best inhibitory effect,and the half maximal inhibitory concentrantion(IC50)was 0.31 mg/m L,compared with the wheat peptide obtained by enzymatic hydrolysis of gluten(IC50was 0.42mg/m L),the wheat peptide prepared from wet gluten was more active;after simulated digestion in vitro,the inhibitory activity of DPP-IV was further enhanced,and the IC50of the digested product was 0.088 mg/m L.The results of cell experiments also proved that wheat peptide and its digested products have a good inhibitory effect on the activity of DPP-IV produced by Caco-2 cells.The relevant active peptides of wheat peptides were determined.After LC-MS/MS analysis and searching the uniprot-Triticum aestivum database,a total of 457 peptides were identified in wheat peptides.On this basis,through computer-aided analysis,using proteomics-related databases and software,the source of the peptide was judged,and the potential biological activity,toxicity,sensitization,intestinal permeability,etc.of the peptide were predicted,and 11 potential biologically active peptides were gained.The three peptides with the highest Peptide Ranker scores were synthesized and verified,and the results showed that QPFP had significant DPP-IV inhibitory activity.The results of molecular docking also showed that QPFP could be linked to residues in the active site of DPP-IV through hydrogen bonds.Peptide related products of wheat were researched and developed.Based on the hypoglycemic biological activity of wheat peptides,the prepared cereal food formula with wheat peptides was obtained through single factor and orthogonal methodology,the optimized conditions were as follows:cooked oatmeal 10 g,wheat fiber powder 5 g,wheat peptide 1 g,whole milk powder 11 g,steviol glycosides 0.012 g,edible sea salt 0.008 g.With this formula,the reconstituted cereal product has comprehensive nutrition and good taste. |
