Study On The Enrichment Technology Of High Fischer Ratio Hemp Peptide And Its Anti-oxidation And Anti-fatigue Evaluation

High fischer ratio peptide is a mixture rich in branched chain amino acids.Due to its unique structure and physiological function,it is widely used in health food and pharmaceutical products.Hemp contains 20-25%protein protein and is a high-quality raw material for preparing biologically active peptides.In this paper,hemp protein isolate(HPI)is used as the raw material to prepare hemp protein hydrolysate(HPH)by optimizing the enzymatic hydrolysis process,and then using activated carbon adsorption,ultrafiltration separation,Sephadex G-25 gel filtration Chromatography technology enriches high fischer ratio hemp peptide(HFRHP),and evaluates its anti-oxidation in vivo and in vitro and antifatigue evaluation.The main findings are as follows:1.Optimal enzymatic hydrolysis process parameters of hemp crude peptide:through the selection of the type of protease and the method of enzymatic hydrolysis,the method of stepwise hydrolysis with alkaline protease and then papain is finally determined.The response surface design was used to optimize the enzymatic hydrolysis process of the two proteases,and the two indexes of hydrolysis degree and peptide content were comprehensively investigated.The optimal process parameters of alkaline protease were:the amount of enzyme added is 10100 U/g,the pH value is 8.5,and the enzymatic hydrolysis at 54℃ for 2 h;the optimal process parameters of papain are:the amount of enzyme added is 5000 U/g,the pH value is 6.5,the enzymatic hydrolysis at 50℃ for 1.5 h.The degree of hydrolysis and peptide content of crude hemp peptide obtained after stepwise enzymolysis were 24.48%and 8.48 mg/mL,respectively,and the yield of crude hemp peptide reached 27.82%.2.High fischer ratio hemp peptide enrichment technology:The best activated carbon model is selected as No.2 activated carbon,and the activated carbon adsorption process conditions are optimized by orthogonal experiments.The best adsorption conditions obtained are:pH value 2.0,carbon liquid the ratio is 1:10,the temperature is 30℃,and the time is 1 h.The OD220/OD280 ratio of hemp crude peptide obtained under the best conditions is 37.58.The prepared hemp peptide was separated by ultrafiltration of 3 kDa and 10 kDa to obtain the component HPH-I(<3 kDa)with the largest OD220/OD280 ratio,and then subjected to Sephadex G-25 gel filtration chromatography to obtain component HPH-I-A,obtained high fischer ratio hemp peptide,its OD220/OD280 ratio reached 47.13,high fischer ratio hemp peptide yield reached 21.65%.Determination of high fischer ratio hemp peptide by gel permeation chromatography(GPC),the molecular weight distribution range from 180 and 980 Da;after amino acid composition analysis,the F value reaches 31.53.3.Evaluation of the anti-oxidation and anti-fatigue properties of high fischer ratio hemp peptide:in vitro anti-oxidation experiments show that when the concentration is 1 mg/mL,high fischer ratio hemp peptide have an effect on DPPH free radicals,hydroxyl free radicals and superoxide.The anion radical scavenging ability is 80.32%,68.02%,61.16%,and the reducing ability is good.The results of animal experiments show that by measuring the antioxidant enzyme system in mice,high fischer ratio hemp peptide can increase the activity of SOD and GSH-Px;the anti-fatigue results show that high fischer ratio hemp peptide has no effect on the body weight of mice,and the three doses are low,medium and high.Compared with the blank group,the weight-bearing swimming time of the group was prolonged by 32.97%,40.52%,and 52.83%,respectively;the lactic acid(LA)content of the mice in the three dose groups after exercise was reduced by 20.44%,39.33%,and 40.67%,respectively;the content of urea nitrogen(BUN)was reduced by 19.39%,39.25%,and 66.54%;the content of malondialdehyde(MDA)was reduced by 19.80%,32.89%,and 42.62%;the content of liver glycogen increased.The correlation analysis of the anti-oxidation and anti-fatigue properties of the high fischer ratio hemp peptide in vivo showed that the SOD activity of the low,medium,and high dose groups of the high fischer ratio hemp peptide is related to LA,BUN,MDA,and liver glycogen.The correlation coefficients were 0.995,0.994,0.942,0.987;the correlation coefficients between GSH-Px activity and LA,BUN,MDA,and liver glycogen were 0.961,0.955,0.989,0.999,respectively,indicating that the high fischer ratio hemp peptide has very good oxidation resistance and fatigue resistance.