Study On The Preparation And Activity Of The Proteolysis Solution Of Dendrobium Spp

Due to having many special physiological functions(anti-hypertensive,anti-oxidation,enhance immunity,etc.),prepared functional peptides which materal come from plant protein is becoming a hot research.Dendrobii Caulis,an endangered precious plant,has been widely used for its good medical effects.In recent years,with the growing shortage of Dendrobium wildlife resources,how to improve the utilization of existing resources Dendrobium becomes one focus to study.The experiment of our group researchers found that the protein content of Dendrobium chrysotoxum Lindl.was about 5.5%,but the studies of appropriate use have not been reported.Therefore,the study of protein based on Dendrobium has important significance.The raw material for this study of the subject is Dendrobium chrysotoxum Lindl..This subject determined the basic component content in three species,the method of extraction and bleaching about Dendrobium protein.Then,selecte specific protease to hydrolyze and to release the active peptide fragments of proteins.We take the obtained small molecules substances to test its corresponding antioxidant and immune activity test.All the work is designed to provide the basis for the rational use of Dendrobium.Through this study,we find out these:(1)The analysis about basic components of polysaccharides、trace elements、protein and amino acid on three kinds of Dendrobium showed that each component has its own characteristics:the polysaccharides and protein content in Dendrobium chrysotoxum Lindl.are in the middle,and they are rich in Sr,Mg,Fe elements;amino acids have high levels;Dendrobium chrysotoxum Lindl.also has a cheap price and a good medical effect,so the choice of Dendrobium chrysotoxum Lindl.as a source of protein material.(2)Select Dendrobium chrysotoxum Lindl.as the raw material to extract protein,determine the different types of protein.Sodium hydroxide concentration,extraction time,Liquid ratio were investigated.The number of factors affecting the extraction rate combined with uniform design method to determine the selected extraction parameters influencing factors within the effective range of each location,the resulting regression equation was:Y=18.9572+0.1760 × X22-0.9018× X1 × X2+0.6307 × X1 × X2-0.0947 × X3 × X4(R2=0.99998,P=0.0067),indicating that this regression equation with high precision and accuracy.Determine the solid-liquid ratio 1:45,extraction time lh,0.3%sodium hydroxide concentration,extraction three times a second,to extract the highest protein content in this condition;liquid extracted proteins activated bleaching experiments investigated the amount of activated carbon,bleaching time,pH,temperature,etc.Bleaching orthogonal experiment to determine the optimum parameters:the amount of carbon is 1.2g,time is 30min,pH is 7.0,when the temperature is 45 ℃.This can obtain the most suitable for decolorization rate and minimal protein loss rate.(3)Explores the alkaline protease,neutral protease,papain,pepsin,trypsin,respectively.Check the protein hydrolysis effect,select neutral protease and papain enzyme complex to process enzyme hydrolysis process.PH,reaction effects of temperature,time,and other factors on the effect of hydrolysis are carried out in this detailed study by design-response surface methodology.This process parameters are studied to establish a regression model fitting equation:Y=18.86+0.31781 × X1-0.072617 × X2-1.43488 × X3+0.32250 × X1 × X2+0.23750× X1 × X3-0.21 × X2 × X3-1.28373 × X12-0.83873 × X22-0.00539683 ×X32(P=0.0074,R2=0.9606).The best ratio of neutral protease and papain is 1:1,the optimum PH is 6.73,hydrolysis temperature is 52.5 ℃,enzyme dosage is 6958U/g,substrate concentration is 4%,hydrolysis time is 3h.Under this condition,the degree of hydrolysis is max.(4)The use of ultrafiltration membrane technology,the hydrolysates are separated different molecular sizes,and greater access to the molecular weight of less than 3kDa and over than 3kDa.The two parts of different molecular weight are tested respectively.With Vc as a positive control,using clear hydroxyl radical,DPPH radical and total antioxidant activity,comparing the size of their in vitro antioxidant activity.The results showed that over than 3kDa had a better antioxidant activity.The tests about the immune activity of hydrolysates are mainly about carbon particle clearance test.The experimental results show that the hydrolysates have a good immune activity.