Research On The Preparation Of Bovine Spinal Peptide And Its Antioxidant Activity

The bovine spinal cord is part of the bovine central nervous system and is often eaten as a highly nutritious food.Chinese medicine believes that it has the effect of medicinal.It is suitable for people with moderate gas,shortness of breath,shortness of body,weak bones,acid and anemia,and yellow eyes.It is rich in protein,fat,arachidonic acid,palmitic acid,stearic acid,linoleic acid,zinc,calcium and so on.The amino acid composition of bovine spinal cord is very close to the needs of the human body,which can enhance the body's disease resistance,and is suitable for people who are conditioned after illness.It can play the role of nourishing blood and repairing tissues,and has great potential for development.However,the current utilization of bovine spinal cord is limited,and the development rate of protein in bovine spinal cord is low.Therefore,this experiment uses fresh bovine spinal cord as raw material,and is separated from bovine spinal cord peptide,by enzymatic hydrolysis by high temperature cooking,beating,degreasing and homogenization.Alkaline protease,neutral protease,papain monozyme and alkaline protease and neutral protease,neutral protein and papain,alkaline protease and papain complex enzyme were used to digest,the degree of hydrolysis and DPPH clearance were the index.The index was optimized for enzymatic hydrolysis conditions,and the bovine spinal peptide was fractionated by ultrafiltration.The antioxidant activity of bovine spinal peptide was studied by in vitro chemical method and cell biological method.Finally,the bovine spinal cord instant nutrition powder was prepared from fresh bovine spinal cord.Below are key research findings;1.The basic components of the fresh bovine spinal cord are as follows;protein15.9%,fat 17.2%,carbohydrate 2.47%,zinc 4.44 mg/kg,calcium 39.7 mg/kg.The proportion of various fatty acids in fresh spinal cord is 21.16%for palmitic acid,23.53%for stearic acid,42.66%for oleic acid,and 12.65%for peanut-ene.It contains all the amino acids the human body needed,and the total amino acid content is 10.51g in 100g of fresh spinal cord.The content of glutamic acid reached 1.77 g/100 g.2.Analysis of bovine spinal protein by SDS-PAGE gel electrophoresis showed that the molecular weight of bovine spinal protein was mainly concentrated at 18.5kDa and there was almost no impurity band.Three kinds of single enzyme enzymatic hydrolysis and three kinds of complex enzyme enzymatic hydrolysis processes were compared with the degree of hydrolysis and DPPH clearance rate.Finally,the two enzymes with good enzymatic hydrolysis,alkaline protease and papain were enzymatically digested,and optimized by orthogonal experiment.Alkaline protease and papain enzymatic hydrolysis process.3.The ratio of alkaline protease to papain was 1:1,hydrolysis degree and DPPH free radical scavenging rate were selected as index,with orthogonal optimization experiment,the enzymatic hydrolysis conditions were enzyme amount was 3%,pH value was 8,hydrolysis time was 3 h,the hydrolysis temperature was 55°C,under the conditions,we got the final hydrolysis degree was 56.2,and the DPPH free radical scavenging rate was 54.67%.4.The bovine spinal cord enzymatic hydrolysate was separated into different peptide components by different ultrafiltration membranes.The in vitro chemical method was used to determine the ABTS free radical scavenging rate,DPPH free radical scavenging,superoxide anion radical scavenging rate and hydroxyl radical scavenging rate.The antioxidant activity was determined as follows:when the concentration of the polypeptide increased from 0.1 mg/mL to 5 mg/mL,the clearance rates of ABTS,DPPH,superoxide anion and hydroxyl radical showed an upward trend,and the ABTS free radical scavenging rate was up to 84.67%.5.Cellular assay showed that bovine spinal peptide had protective effect on H₂O₂-induced HepG₂ oxidative damage.Bovine spinal peptide treatment significantly increased the concentration of superoxide dismutase SOD and GSH and significantly decreased the content of MDA in cell.6.The bovine spinal cord instant powder was prepared by spray drying method,and the drying process was optimized.The optimal spray drying conditions were determined as:inlet air temperature was 185°C,feed amount was 20 mL/min,soluble solids was 14%,and 25%of?-cyclodextrin addition.