| Pine seeds are the seeds of pine trees,also known as pine kernels,sea pine seeds,pine nuts,pine seeds,and pine nuts.They are the edible seeds of plants of the genus Pinus(Pinus L)of the class Pinus L.Our country has very rich pine nut resources.In the northeast,Korean pine is the main source.The nutrient value of Korean pine seeds is extremely high.The main nutrients are oil and protein,as well as a small amount of trace elements and vitamins.Pine nut protein has unique nutritional properties and is a plant protein resource that has not yet been fully developed but has extremely high application value.Therefore,in this study,pine nut meal was used as the raw material,and the pine nut protein was first extracted by the method of alkaline extraction and acid precipitation.Secondly,it is enzymatically hydrolyzed to obtain pine nut peptide.Sephadex G-25 gel chromatography was used for separation and purification,and the molecular weight was determined to identify its amino acid composition.On this basis,the obtained pine nut peptides were studied for anti-fatigue,anti-oxidation,and hypolipidemic activities.The specific research content is as follows:1.Using Korean pine pine seed meal as the raw material,the pine seed protein was extracted by alkali extraction and acid precipitation,and the extraction process was optimized by single factor combined with orthogonal experiment method,and the optimal reaction time was 2h and the optimal temperature was 55.?,the optimal pH value is 9,the optimal material-liquid ratio is 1:25,and the optimized extraction rate of pine nut protein can reach 58.34%.2.Use neutral protease,alkaline protease,papain and trypsin to study the enzymatic hydrolysis of pine nut protein,and compare the degree of pine nut proteolysis by different proteases.The results showed that alkaline protease had the best enzymatic hydrolysis effect,so alkaline protease was used to optimize the enzymatic hydrolysis process of pine nut protein.Through the single factor and response surface method,the optimal enzymolysis temperature is50?,enzymolysis time is 2 h,pH value is 9,the amount of enzyme added is 8100 ?/g,and the substrate concentration is 2%.Under this technological condition,the degree of proteolysis can reach 34.87%;the pine nut peptide is separated and purified by Sephadex G-25 gel chromatography to obtain four components,named PP-1,PP-2,PP-3 and PP-4,the smaller molecular weight parts of PP-3 and PP-4 are combined and named PP-5,and the molecular weight of PP-5 is determined to be between 65 and 3605 Da by liquid chromatography.Further analysis of the amino acid composition of PP-5 revealed that pine nut peptide is rich in arginine,glutamic acid,alanine,valine,tyrosine,etc.Among them,arginine content accounted for 15.43%,glutamic acid content accounted for 8.28%,and alanine content accounted for 5.88%.3.In the study of anti-fatigue activity,Kunming mice were selected as the research objects to be divided into groups,which were set as blank control group,positive(whey protein)control group,and pine nut peptide(25% peptide solution/d,50%).Peptide solution/d,100% peptide solution/d)dose group.Mice in each group were given intragastric administration for 6 weeks and simultaneously performed 6-week swimming training.After the last training,the weight-bearing swimming time of the mice was recorded,and the serum lactate dehydrogenase,blood urea nitrogen,blood glucose levels and the mice were measured.Glycogen content in muscle tissue and liver tissue.The results showed that compared with the blank group,the content of lactate dehydrogenase in the middle and high dose groups of pine nut peptide was significantly increased,and the content of muscle glycogen,liver glycogen and blood sugar levels were significantly increased,indicating that pine nut peptide can effectively reduce Muscle glycogen and liver glycogen were consumed to maintain normal blood glucose levels in mice,and blood urea nitrogen was also significantly reduced.The above results can prove that pine nut peptide has a good anti-fatigue effect.4.In the study of antioxidant activity,D-galactose was used to establish a mouse oxidative damage model.Except for the blank control group,mice in the other groups were injected intraperitoneally with 5% D-galactose.The MDA content increased significantly,proving that the model of oxidative damage in mice was successful.On this basis,a positive control of Vc was set up,the pine nut peptide(25% peptide solution/d,50% peptide solution/d,100% peptide solution/d)dose group,and the mice were gavaged continuously for 6 weeks,and each group was determined Compared with the model group,the activities of GSH,T-AOC,CAT,SOD in the mouse serum and liver of the dose group were significantly or extremely significantly increased,and the content of MDA was also significantly decreased,indicating that pine nut peptide can effectively inhibit the oxidation of mice Injury: The results of analyzing the pathological sections of the liver of mice showed that compared with the model group,the pine nutin low,medium,and high dose groups had improved liver cell damage in different degrees.The above research results show that pine nut peptide has a good antioxidant effect and can effectively alleviate the body's oxidative damage.5.Study on lipid-lowering activity: First,the mice were fed with high-fat diet for 4 weeks.The results of morphological and serological tests showed that compared with the normal group,the liver tissue of the high-fat model group was locally fatty.,And the total cholesterol in the serum also increased significantly,indicating that the modeling was successful.Then set the dose group(25% peptide solution/d,50% peptide solution/d,100% peptide solution/d)for a period of four weeks.After feeding for 4 weeks,the serum TC,TG,HDL in each group of mice were measured-C and LDL-C indicators,the results show that the administration of pine nut peptide can significantly or extremely significantly reduce the content of TC,TG,LDL-C in the serum of mice,and can significantly or extremely significantly increase the content of HDL-C Among them,the pine nut peptide high-dose group has the most obvious effect.The above results show that pine nut peptide has a good effect on lowering blood lipids. |
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