The Preparation Of Corn Antihypertensive Peptide And Simulated Moving Bed Chromatography Separation Technology

In order to strengthen the utilization rate of corn protein powder,improved its added value.In this experiment,took the corn protein powder(protein content 70%)as raw materials.through pretreatment?hydrolysis technology?extraction and purification technology,to simulated moving bed chromatography separation of industrialization,finally got higher activity corn antihypertensive peptide.The main research contents and conclusions are as follows:In the comparative experiments between composite enzyme and single enzyme,the two kinds ways' conclusions were almost similar in ACE inhibition rate,integrated consider,chose single enzyme AS.1398 neutral protease for best hydrolysis enzyme and carried out the optimization of enzymatic hydrolysis conditions.Used single-factor experiment and orthogonal test to carry out the optimization of enzymatic hydrolysis conditions.Chose the pH value,temperature,substrate concentration,the amount of enzyme [E] / [S] of the single-factor,and the four factors to determine the parameter values L9(34)orthogonal test,through ACE inhibition rate in vitro testing and yield of peptide indicators to determine the optimum technology conditions.The result indicated that AS.1398 neutral protease was the best hydrolytic enzyme,when the hydrolysis time in 2h,pH 7.0,temperature 50 ?,5% substrate concentration,the amount of enzyme [E] / [S] 1.5:100,and the maximum ACE inhibition rate was 88.25%,the yield of peptide was 59.58%.In the steps of extraction and purification technology,firstly used ultrafiltration segregation,got the peptides which molecular weight below 1000 the ACE inhibition rate is higher than others,and then used sephadexG-15 to separate again,chose distilled water to elution and got four samples,after the examination,the sample 1 was better,its ACE inhibition rate reached to 91.21%,is was the target product,by HPLC examination,the purity and activity is higher,and the molecular weight at about 950 Da~1420 Da.After macroporous resin adsorption separation,in static adsorption and desorption examination,screened the XAD761 type resin for the best from four resins,not only its higher adsorption,but also its higher desorption rate,then through different pH value of the sample in the static adsorption test and different concentrations of desorption solvent in the desorption test,finally got the conclusions:pH6.0,desorption solvent(alcohol)concentration was 70% for the best;In dynamic adsorption and desorption examination,finally got XAD761 resin separation technology parameters of the corn antihypertensive peptide: the saturated adsorption was 17 mg/mL;the speed of feed material was 3.0 mL/min,the speed of water elution was 1.5 mL/min,the speed of 70% alcohol elution was 3 mL/min,the amount of elution solvent was 280 mL,and the speed of water material was 2.0 mL/min,the system has no alcohol.And compared ACE inhibition rate between the preseparation and the after separation,took the after separation ACE inhibition rate was 90.35%,further shows that the XAD-761 resin can did well in extract and purify corn antihypertensive peptide,and raise corn antihypertensive peptide extraction,segregation,and enrichment efficiency,provided a good data base for the next step of the industrialization of the separation.In the simulated moving bed chromatography separation technology test,which separated the higher activity corn antihypertensive peptide,ascertainment the concentration of sample was 16 mg/mL,the speed of the sample,the water elution1,the alcohol elution and the water elution2 discern as: 15.0 mL/min,20.0 mL/min,10.0 mL/min,26.0 mL/min,by three times repetition and verification experiment.The result was that the purity of corn antihypertensive peptide was 95.0%and the yield of it was 93.1%,its ACE inhibition rate was 93.8%.By HPLC detection analysis,the higher activity corn antihypertensive peptide,which molecular weight scope was 700 Da ~ 900 Da,and corn antihypertensive peptide's molecular weight for 776 Da accounted for more than 68%.And make use of the pepsin and chymotrypsin in simulation experiment,the experimental results show that,the higheractivity corn antihypertensive peptide can tolerate the effects of the pepsin and chymotrypsin in gastrointestinal tract,had strongger stability and the function of antihypertensive was better.