Preparation And Immunologic Function Research Of Pine Nut Polypeptide

Pinus koraiensis Sieb.et Zuee is a rare tree species of Xiao xing an ling in China.The pine nut is a high nutritional value of fruits,rich in protein,amino acids,fatty acids,vitamins, minerals and other substances physical activity has a wide application development potential.In this research,pine nuts were used as material,the use of modern analytical tools from the pine nut protein extraction technology,hydrolysis condition optimization,pine nut polypeptide molecular weight distribution estimation,as well as estimates immunologic functions of the pine nut polypeptide(PNP).The main results are as follows:1.Respectively alkali extraction method and ultrasonic extraction from pine nut protein, optimum conditions were:NaOH0.10mol/L,material and fluid compared 1:14,the extraction time was 90min under 50℃;ultrasound treatment for 15min at power of 900W,liquid ratio 1:10.Two methods of pine nut protein extraction rates were 67.45%and 97.65%.Ultrasonic extraction method can be effective in raising the pine nut protein extraction rate,shorten response cycle.Have a highly-efficient,energy-saving advantages.2.Select Alcalase proteinase,the neutral proteinase and the trypsin as a pine nut protein enzymolysis preparation enzyme,the enzyme effect pine nut protein research,in accordance with its hydrolysis of Alcalase identified as the best use of protease enzyme.In the role of this enzyme hydrolysis under optimal conditions:hydrolysis temperature is 50℃,pH 8,150 min hydrolysis time,the enzyme amount of 5000U/g,substrate concentration of 8%.Under this condition,the pine nut protein hydrolysis of the 30.06%.Its enzyme substrate concentration has significant impact,followed by the enzyme dosage.3.The crude pine nut polypeptide(PNPC)by Sephadex G-25 gel-filtration chromatography purification,has three main fractions,respectively PNP1～2,PNP3 and PNP4～7.Select several proteins of low molecular weight standards,the use of this method of molecular weight distribution,the three main points-roughly the molecular weight distribution:more than 23105Da,3776Da about and less than 249Da.According to the definition of bioactive peptides, select the highest activity PNP3 components research,type and content of amino acid analysis and conjecture the purified pine nut polypeptide(PNPP,namely PNP3) has immunnologic function.4.The PNPP immunologic function of the confirmatory tests,the experimental results showed that the dose can be significantly increased the weight of immune organs in mice,the cells of mice immune function,humoral immune function and monocytes - macrophage phagocytos is also have a catalytic role in immune regulation that PNPP a role.