Study On Improving Sleep Effect Of Oyster Enzymatic Hydrolysate And Product Development

Sleep is an essential physiological activity of the human body,which can relieve fatigue and restore the spirit,but sleep problems will affect human health.In recent years,drug treatment and health food are still used to treat sleep problems,and longterm use of drugs for insomnia will lead to drug addiction and other side effects.Oyster is one of the main economic shellfish in artificial culture in China,which is rich in nutrients needed by human body and an important resource for the study of active substances.In addition,<Treatise on Febrile Diseases> records show that Bupleurum and Dragon Bone Oyster Soup can play a calming and soothing role.Therefore,this paper takes Crassostrea hongkongensis as raw material and uses controlled enzymolysis technology to prepare oyster enzymatic hydrolysis product.Through animal experiments to verify and screen the best active components to improve sleep,and develop sleep-improving products,it provides theoretical basis and practical direction for subsequent mechanism research and development of related functional products.The research contents and results of this paper are as follows:(1)The enzymatic hydrolysis product of oyster(EPO)was prepared by animal protease and determine its basic physical and chemical components;the way of gavage,the sleep improvement effect was evaluated by direct sleep test,extended pentobarbital sleep time test and other animal behavioral experiments,and the levels of inhibitory neurotransmitters and excitatory neurotransmitters in the brain of mice were measured.The results showed that the content of the molecular weight distribution of EPO is mainly concentrated in the range of <3 KDa,EPO protein was 56.83% and the amino acid composition and mineral element composition was rich.Compared with the negative control group,EPO had no direct hypnotic effect,EPO can improve sleep mainly by prolonging sleep time,increasing sleep incidence,shortening sleep latency,increasing the contents of three inhibitory neurotransmitters,GABA,5-HT and MT,and reducing the contents of two excitatory neurotransmitters,DA and NE in the brain of mice.Therefore,this study continued to use the way of intragastric administration,through animal experiments and neurotransmitter content,nerve cell markers protein expression and other physiological and biochemical indicators for the next step to improve sleep the best active components screening and mechanism.(2)The enzymatic hydrolysis product of oyster(EPO)were fractionated by ultrafiltration to obtain <3 K,3-5 K,>5 K three ultrafiltration components,and determine its basic physical and chemical components;continue to use animal behavior experiments to improve sleep activity evaluation and component screening;through the mice weight experiment,organ coefficient experiment to verify the safety of ultrafiltration components;through the determination of mouse brain neurotransmitter content and GFAP expression to explore its mechanism of improving sleep.The results showed that the molecular weight distributions of <3 K,3-5 K and >5 K were mainly in the range of <3 KDa,the protein contents of <3 K,3-5 K and >5 K were 76.39%,81.98% and 61.35%,respectively.The amino acids were complete,and they were rich in zinc,iron,magnesium and other mineral elements related to sleep.UEPO had no effect on body weight and organ coefficient of mice.In terms of behavior,the three ultrafiltration had no direct hypnotic effect,UEPO can prolong sleep time,increase sleep incidence and shorten sleep latency.In terms of neurotransmitter content,UEPO can increase the contents of GABA and 5-HT inhibitory neurotransmitters and decrease the contents of DA and Glu excitatory neurotransmitters in the brain of mice.UEPO can also significantly inhibit the expression of GFAP in the brain.Comprehensive comparison,screening <3 K components to improve sleep effect is the best.Therefore,this study will focus on <3 K component for the preliminary development of sleep improvement products.(3)Take oyster enzymatic hydrolysis product <3 K as the main material,the compound ratio of Semen Ziziphi Spinosae,Lily and honey powder was selected to improve sleep function,and the best compound ratio of flavor and stabilizer was obtained by response surface methodology.The results of formula optimization were as follows: 5.00% < 3 K oyster enzymolysis powder,1.00% jujube seed powder,1.00%lily powder,1.00% honey powder,7.00% apple powder,0.40% stevioside,2.00% ?-cyclodextrin,0.15% xanthan gum,0.10% pectin,0.20% CMC-Na.Under the optimized conditions,the product has pleasant fragrance,uniform texture and no precipitates,and the contents of heavy metals and microorganisms meet the national food standards.