Preparation Of Soy Protein Hydrolysate And Its Application In Cell Culture

Soy contains about 40%protein,which provides the eight essential amino acids.This study use double enzyme Hydrolysis of soy protein to prepare its hydrolyzate and test its physical and chemical properties.Then using it in cell cultrue.This study provide a theoretical basis for the application of soy protein hydrolysate in cell culture.Firstly,the paper are described background and status of protein hydrolyzates,plant protein hydrolysates,preparation of soy protein hydrolysate and followed by the hydrolysis of soy protein by double enzymatic hydrolysis to prepare the soy protein hydrolyzate.Twokindsofnon-animal-derivedprotease(papain,alkaline protease,compound protease and neutral protease)were used in hydrolysis.The degree of hydrolysis was taken as an index,And the degree of hydrolysis of four kinds of enzymes in different proportions was tested in combination with each other,and the optimal enzymes for hydrolysis were selected.On the basis of single factor test,the orthogonal test was used to optimize the enzymatic hydrolysis of soy protein.The enzymolysis product was added into the medium to detect the growth of animal cells.The results showed that the optimum hydrolysis conditions were as follows:temperature 60?,pH value 7.0 and enzyme/substrate(E/S)=1:4(Mass ratio),hydrolysis time of 4 h,the degree of hydrolysis of 43.7%.Testing of the physical and chemical properties showed that the soy protein hydrolysate has good solubility and good thermal stability at the pH values of 2,4,6,8 and 10.The prepared protein hydrolyzates also has a good foaming which are easy to dissipate.So it will mechanical damage of cells if you add it into the medium.The study also made free amino acid detection of soy protein hydrolysate.The results show that it contains a large number of amino acids,especially the concentration of glutamine.It is contains2.038%.By measuring the osmotic pressure of the mediumwhich added soy protein hydrolyzates,there was no significant difference between the medium and the medium without hydrolyzate.Finally,we reaserched the cell subculture,growth curve and cell metabolism in the cell culture by adding soy protein hydrolysates in medium.The results showed that:Soy protein hydrolysates can stimulate the growth of SP20,CHO cells.By MTT assay,the soy protein hydrolysate was added to the culture medium with 0.25%concentration.And the two cells were stable for more than 15generations.In addition,cell morphology did not change in the process.During the cell culture,the maximum density of SP20 and CHO cells were respectively 14.80×10~5 cells/mL and 13.89×10~5 cells/mL which are higher than the control cells.Their cell double time(DT)was respectively 18.1 h and 18.6h.During the metabolism test,it is found that cells in the form of more energy efficient use of glucose with soy protein hydrolysate compared with the control group.And the accumulation of lactate acid slightly higher than the control group.It is good at the synthesis of other biological molecules.In this paper,we studied the optimization of the preparation process of soy protein hydrolysate,the detection of physicochemical properties and the research on the growth of cells.It is found that soy protein hydrolysate can promote cell growth.And it also provides a theoretical basis in cell culture and serum substitute,which has a good prospect for development and application.