Study On The Production Process Optimization And Sustained Release Of Egg White Peptide Liposome

This research is based on Jilin Science and Technology Development Program(20180520045JH).China is the world’s largest egg production country,and the egg processing industry occupies an important position in the national economy.At present,China’s egg processing industry is still dominated by primary processing.Although the demand for refined products is gradually increasing,the corresponding technical reserves are insufficient.Eggs are rich in high-quality protein,and are an excellent source for the preparation of bioactive peptides such as antioxidant peptides,and have a high value of deep processing.However,egg white antioxidant peptides and other active peptides have problems such as insufficient gastrointestinal stability,easy degradation in the body,and low bioavailability,which seriously limits the development and application of related products.In order to improve the stability of active peptide products,this paper takes egg white antioxidant peptides as an entry point,optimizes the production process of egg white antioxidant peptide liposomes by constructing egg white antioxidant peptide liposome delivery systems,and passes in vitro/in vivo experiments Exploring its slow-release effect,we finally obtained an egg white antioxidant peptide liposome with a smaller particle size,a higher embedding rate,and a better sustained-release effect,which provided a certain degree for the development of egg white antioxidant peptide liposome products.Technical Support.The main research results of this paper are as follows:(1)Using ethanol injection-high pressure homogenization technology,egg yolk lecithin,cholesterol,etc.were used to prepare the liposome wall material.The particle size and PDI were used as the measurement indicators,and the liposome was determined through a combination of single factor and response surface experiments.The best preparation method of the wall material is:egg yolk lecithin concentration 11mg/mL,high pressure homogenization pressure40 MPa,high pressure homogenization time 7.5 min,the particle size of the liposome wall material obtained under this condition is 89.4±2.4 nm.By comparing the effects of five different stabilizers on the liposome wall material particle size and PDI,the best stabilizer was selected as Tween80.(2)Enzymolysis of egg white protein powder with alkaline protease to obtain egg white antioxidant peptide EWPH.Using ultrafiltration and nanofiltration to separate and purify,the egg white antioxidant peptide components EWPH1,EWPH2,EWPH3 with molecular weights of 200Da~1 kDa,1 kDa~3 kDa and 3 kDa~5 kDa were obtained.The antioxidant activity of each component was measured separately.The EWPH1 egg white antioxidant peptide component(molecular weight 200 Da~1kDa)has the strongest ABTS~+free radical scavenging rate,ORAC free radical scavenging rate,and FRAP free radical scavenging rate.The core material was subjected to subsequent embedding experiments.(3)The embedding rate and particle size are used as indicators to optimize the embedding process of egg white antioxidant peptide liposomes.Through the combination of single factor and response surface experiments,the optimal preparation conditions of egg white antioxidant peptide liposomes are determined as:The ratio of the core material wall material is 3:5,the ultrasonic time is 45 s,and the ultrasonic power is 150 W.Under these conditions,the embedding rate of the egg white antioxidant peptide liposomes was 65.7±3.6%.(4)In vitro and in vivo sustained release experiments were conducted to investigate the active substance delivery and sustained release capabilities of egg white antioxidant peptide liposomes.In vitro sustained-release dialysis bags were used to construct simulated gastric fluid(SGF)and simulated intestinal fluid(SIF)environments.In the simulated gastric fluid,the free peptide group was released into the medium faster than the liposome group,and the release rate reached 64±7%at 2h.At this time,the release rate of liposome peptide was only 48±6%.In simulated intestinal fluid,the release rate of liposome peptides was always lower than that of free peptides at the same time.Egg white antioxidant peptide liposomes have a good in vitro simulated gastric and intestinal sustained-release effect.(5)In vivo sustained release using HPLC-MS technology combined with Fmoc-Cl pre-column derivation,using blood lysine,tryptophan,phenylalanine,threonine,valine and other essential amino acids as detection markers,Indirectly investigated the sustained release of egg white peptide in mice.Compared with the egg white peptide group and the blank control group,the peak value of various amino acids in the egg white peptide liposome group was significantly delayed,and the concentration was significantly improved.It can be seen that while the egg white peptide exerts a sustained-release effect,it may have a certain effect of promoting the absorption of egg white peptide.