Research On The Key Technologies Of Immune Corn Bran Polysaccharides

This study was supported by Changchun Science and Technology Plan Program?13NK03?.Using corn bran as the raw material,four main tests about CBPS were carried out including extraction technology,purification and structure,quality control,research of immune activity in vitro.?1?The efficient extraction technologies of CBPS were researched.One-factor-at-a-time?OFAT?and response surface metrology?RSM?experiment were used to optimize the conditions of tests.Extraction rate of CBPS was selected as the measurement index,four different technologies including hot water extraction,cellulase extraction,pulsed electric field?PEF?assisted extraction,electron bean irradiation?EBI?assisted extraction.The results were obtained about hot water extraction,it showed that the optimum extracting process were as followed:extraction time of 3.5 h,extraction temperature of 70 ?,solid liquid radio of 1:35.On these conditions the extraction rate was 7.81±0.34%.The regression model of cellulase was Y=13.99-2.32X₁-0.43X₂+0.15X₃-0.03X₁X₂+0.41X₁X₃-0.14X₂X₃-5.01X₁²-1.63X₂²-0.38X₃²,R²=0.9979,which got a result of 13.8±0.10 %.In the study of EBI,the result was the best when the irradiation dose got to 108.7 k Gy.The regression model of PEF was Y=15.54+0.37 X₁+0.074 X₂+0.46 X₃+0.56 X₁X₂+0.063 X₁X₃+0.17 X₂X₃-0.77 X₁₂-0.84 X₂₂-0.84 X₃₂,R²=0.9853,the optimum parameters were electric field intensity of 25 k V/cm,electric field frequency of 2080 Hz,liquid solid radio of 42:1 m L/g.At the same time,the material was scanned under scanning electron microscope?SEM?.With the increase of the irradiation dose,the surface of the material was more and more rough.?2?The separation and purification of CBPS and structure of a preliminary study were researched.Using AB-8 macroporous adsorption resin,the static adsorption and dynamic adsorption combination of decoloring and deproteinization.Optimum process parameters of static adsorption,respectively,the water extraction process of polysaccharide,1.5 h,40 ?,liquid column on pH 6.0,liquid mass concentration on column 3 mg/m L;Enzyme extraction process of polysaccharide: adsorption time 2.0 h,adsorption temperature 40 ?,the column liquid pH 6.0,liquid mass concentration on column 4mg/m L;Electron beam irradiation assisted enzymatic extraction polysaccharides: 2.0 h,adsorption time,adsorption temperature 45 ?,liquid column on pH 6.0,liquid mass concentration on column 3 mg/m L;High-voltage pulsed electric field assisted enzymatic extraction polysaccharides: 1.5 h adsorption time,adsorption temperature 45 ?,liquid column on pH 6.0,liquid mass concentration on column 4 mg/m L.Four methods to extract polysaccharides had a same characteristic on CH3,CH2,CH stretching vibration,and C=O stretching vibration,bending vibration of C-H.The number average molecular weight of four methods respectively 84589 Da and 92968 Da,103462 Da,135637 Da,173564 Da,173564 Da.?3?The change of the water as the research object,with the help of LF-NMR technology.Polysaccharide contained in three different states of water components,combined water,immobilized water,free water.Determination of trace 48 h,water in CBPS increased under 25 ?,and had a dynamic change under 4 ?.?4?Immune activity in vitro studies by the method of cell biology,by Griess method of NO release quantity for testing,establishing standard curve: Y = 0.0143 + 0.0671 X,R²=0.9957,good linear relationship.With the increase of polysaccharide concentrations of NO release a quantity to increase,when the polysaccharide concentration is 800 ?g/m L NO release a quantity to achieve maximum 47.93±1.76 microns,and different methods to extract the polysaccharide difference is not significant.Using the method of ELISA inspection on production of TNF alpha,build up the standard of binary quadratic curve model is: Y=0.2662-10^-6 X² + 0.003 X,R²=0.9988,the fitting relationship is better.With the increase of polysaccharide concentrations,the concentration of TNF alpha increases,when the polysaccharide concentration reached 800 ?g/m L concentration of TNF alpha best 725.79±4.21 pg/m L,and different methods to extract the polysaccharide differences are not significant.