Study On The Preparation Of Polypeptides From The Yellow Skin Of Anser Cygnoides’ Palm And The Application In Antifreeze Moisturizing Cream

The yellow skin of wild goose palm is that the yellow skin on the flaking foot and webbed of the Anser cygnoides.The contents of Compendium of Materia Medica are as follows: " Baking,oil blending,frostbite good".In this paper,in order to develop its application in anti-freezing products,the preparation of peptides,anti-low temperature damage activity and its application of cream were studied by using the yellow skin of Anser cygnoides’ palm as raw material.The results are as follows:Through single factor and orthogonal experiment,the best extraction process of the protein from the yellow skin of Anser cygnoides’ palm(YSPR)was determined.Enzymatic hydrolysis process of the protein from YSPR was optimized by response surface method;the peptide was purified by molecular exclusion chromatography;the amino acid composition was determined by amino acid analyzer;it was analyzed by UV absorption spectrum scanning and UPLC-Q-TOF-MS/MS.The different components of the polypeptide were separated by gel chromatography.The results showed that the optimal extraction conditions were as follows: ultrasonic temperature 35 ℃,ultrasonic power 400 W,ultrasonic time 40 min,solid-liquid ratio 1:35.After screening neutral protease as a tool enzyme,the optimal enzymatic hydrolysis process of the protein from the yellow skin of Anser cygnoides’ palm was optimized by response surface method: under the conditions of enzymolysis time of 5 h,enzymolysis temperature of 55 ℃,p H7.0,and enzyme dosage of 4200 U/g.Using molecular exclusion chromatography to separate the polypeptides from the protein from yellow skin of Anser cygnoides’ palm and the molecular weight of the polypeptide was in the range of 859-10302 Da.The polypeptide contained 17 kinds of amino acids,with essential amino acids accounting for 42.27%.The polypeptide was scanned by ultraviolet absorption spectrum,and there were two absorption peaks at 220 nm and 280 nm.The results of UPLC-Q-TOF-MS/MS showed that it was composed of mixed peptides.Three components YSP1,YSP2 and YSP3 were separated by G-50 dextran gel chromatography.To investigate the anti-low temperature damage activity of vascular endothelial cells(VECs)in rats by establishing a low temperature injury model to detect the changes of ET,NO,SOD,MDA,cytokines in VECs at low temperature by observing the cell proliferation of human fibroblasts(HDF)and applying scratch experiment,the skin healing ability was investigated,and the moisturizing activity was investigated by using discoloration silica gel.The results showed that the four components of the protein from YSPR and polypeptide the yellow skin of Anser cygnoides’ palm YSP1 、 YSP2 、 YSP3 all had protective activity against low temperature damage VECs,when the YSP3(400 μg/m L),the protection rate of low temperature injury VECs is the largest.The effect on vasodilation factors was extremely significant(P<0.01);the effect on the secretion of cytokines was very significant(P<0.01).From many aspects,it shows that the four components of the yellow skin of Anser cygnoides’ palm can protect the VECs of low temperature damage.The four components of the yellow skin of Anser cygnoides’ palm have the function of promoting skin healing.Among them YSP3 function is strongest,when the concentration was 400 μg/m L HDF the cell proliferation rate and wound healing rate after 24 h were135.39±7.03% and 53.61±1.59%,respectively.The four components of the yellow skin of Anser cygnoides’ palm have moisturizing effect,among them YSPR moisturizing effect is strongest,have a moisturizing effect at 5 h,moisturizing rate is 32.6±0.56%.The YSP3 group was divided into raw materials to prepare antifreeze moisturizing cream.The safety and irritation of the cream were investigated by oral test and skin irritation test in mice,and the stability and sensory application of the cream were evaluated.The results showed that there was no death or poisoning in the oral test.In the skin irritation experiment,no adverse symptoms such as redness,swelling,rupture and so on appeared,thus proving that the antifreeze moisturizer is non-toxic and non-irritating.It performs well in cold and heat resistance experiments,without precipitate and p H is 7.0,which meets the industry standard.In the sensory evaluation,the appearance,ductility,moisture and tenderness of the cream were all good,and the skin was delicate and moist after using the cream.