The Study On Exteaction And Physiological Function Of Protein In Korean Pine Seeds

Korean pine(Pinus koraiensis Sieb.et Zucc) is rare species of trees mainly produced in China Xiaoxing'anling.Its Seeds have good functions of softening blood vessels,lowering blood lipids,cholesterol,triglyceride and preventing aging,so it has broad potential in application and exploitation.Protein is an important component of various biological organisms including humans, which is one of the material bases of human life.The growth,development,genetics, reproduction,and other life active substance of organisms can not be separated from protein.In some of the physiological activity of substances,such as peptide hormones,antibodies, enzymes and nucleoprotein are all proteins,they play a very important role in regulating physiological functions,maintaining the metabolism.Protein is everywhere in the life activities, and different proteins have different structures and functions.Pine nut contains higher protein of 13%-20%,the amino acid composition of pine nut is complete,and the essential amino acids are generally higher than plant protein.Glutamate in pine nut is the highest content of 16.3%.Glutamate can reduce blood ammonia at the clinic,be used for the treatment of memory,language barriers,such as children with mental incompetence caused by schizophrenia and cerebrovascular disorders.Pine nut can be considered as hyperactivity nourishing heart to calm mind for these causes in traditional medicine.The seed of Korean pine is regarded as the raw materials in this paper,and makes a research and comparison on extraction technology of pine nut protein by utilizing modem analysis means of separation in order to elicit the best technology condition of extraction.And then the effect of pine nut protein on the memory function,immune function,anti-fatigue and other physiological functions of mice have been studied.The results are as follows:1.The extraction methods of pine nut protein are Water Extraction and Ultrasonically Assisted Water Extraction.In the experiment of the hot water extraction of pine nut protein,the optimum extraction technology is obtained by single factors with orthogonal test:solid-liquid ratio is 1 to 15(m/v),extraction time is 150min,extraction temperature 40℃.At the optimum extraction technology,the extracted protein quality is 0.67g and extraction rate is 26.3%by verification test.In the experiment of Ultrasonically Assisted Water Extraction,the extraction condition has been optimized by the method of single factors binding response surface and the best process is that solid-liquid ratio is 1 to 36(m/v),extraction time is 128min,extraction temperature 37℃.The extracted protein quality is 0.67g and extraction rate is 26.3%at these conditions.The experiment shows that the method of ultrasonic-assisted extraction can improve the extraction rate of pine nut protein from 26.3%to 69.4%by single factors with response surface methodology.2.The extracted pine nut protein is used to conduct functional experiment on anti-fatigue function,immune regulation function,memory function of mice.The experiment shows that:The result of mice of irrigated with pine nut protein in comparison with control group shows that the pine nut protein has no significant impact on the weight of mice(P＞0.05),Low-dose group has significant impact on carryed swimming time of mice(P＜0.05) and the middle and high-dose group is significant(P＜0.01);Low-dose group has very significant impact on the content of blood urea nitrogen of mice(P＜0.01) and the middle and high-dose group is significant(P＜0.05);These results suggest that the pine nut protein can improve the anti-fatigue ability of mice.The result of mice of irrigated with pine nut protein in comparison with control group shows that the pine nut protein has no significant impact on the weight,thymus index and spleen index of mice(P＞0.05),Middle and high-dose group has significant impact on the toe thickness of mice(P＜0.05) and the low and middle-dose group has significant impact on quantitative hemolysis value(P＜0.05),high-dose group has very significant impact(P＜0.01).These results suggest that the high-dose of pine nut protein has very significant impact on the enhancement of mice's immunologic competence(P＜0.01).The result of mice of irrigated with pine nut protein in comparison with control group shows that the pine nut protein has no significant impact on the step down-experiment and the content of malondialdehyde in mice brain tissue(P＞0.05),Middle and high-dose group has significant impact on AChE in mice brain tissue(P＜0.05).These results suggest that pine nut protein can decrease the content of AChE in mice brain tissue and has certain regulation to the learning and memory function of mice.The experiment of pine nut protein on the physiological function of mice shows that:the pine nut protein can enhance the capacity of anti-fatigue,the immunity of mice and improve mice's ability of learning and memory.