The Study Of Rapeseed Protein ACE Inhibitory Preparation And Seperated

Hypertension give priority to with increased blood pressure and is the important cause of cerebrovascular diseases and its risk factors. Common blood pressure medications have many adverse effects. It forcing people to continuously improve for more safe blood pressure medications. Rapeseed is one of main oil-bearing crops in our country. Rapeseeddregs is widely regarded as feed of animals, skim rapeseed contain 30%-40%protein. It is a potential, ideal for high quality protein resources and the raw material of antihypertensive peptides.The test established RP-HPLC of the determination on ACEI of rapeseed protein in vitro activity detection. The result showed that the time of reaction was 50 min and the rapeseed protein hydrolysate had to deal with ethyl acetate.The method was further validated by captopril.Takeing rapeseed protein as the raw material, comparing the alkali protease, neutral protease, acid protease and pineapple proteinase in its hydrolysis effect, alkali protease was selected, because it get solution with higher enzyme hydrolysis degree and ACE peptide inhibitory activity by alkali protease. And its hydrolysis degree and inhibitory activity 14.25% and 53.30% respectively.And with alkali protease for hydrolysis enzyme,The effects of concentration of substrate,hydrolysis time,[E],temperature and initial pH value added degree of hydrolysis and the ACE peptide inhibitory activity were studied. The optimal conditions were that concentration of substrate 4%,time 3h, [E] 6AU/g,reaction temperature 50℃,the pH range 9. Based on these, the experiment of ultrasonic pretreatment of rapeseed protein was carried out. Through Response Surface Analysis (RSA),the results showed that the optimum conditions of ultrasonic pretreatment were 23.5min of pretreatment time,202.8 W of ultrasonic power and 49.5℃of temperature.In order to remove macromolecular proteins, ultrafiltration was investigated with lOkDa ultrafiltration membrane. The results show that the optimum conditions of ultrafiltration were 12g/L of concentration of solvent, and 200KPa of pressure Active fraction was separated by gel. The effect of eluent was studied. The results showed that water is good eluent and get 3# with the highest ACE inhibited activity. The 3# peak's sample was separated by HPLC. The gradient elution conditions of mobile phase was decided, c# peak is the sample with the highest active, more than 95% purity and 0.0118 mg/mL IC50.The stability of rapeseed protein hydrolysate were studied. Angiotensin-I-converting enzyme inhibitory pepetides of rapeseed decreased in the inhibition of effect to ACE with temperature rise and the time extension. And the results of intestinal enzyme action showed that digesting of rapeseed protein hydrolysate around the active were almost the same, and approved by HPLC experiment. It explained that the rapeseed protein hydrolysate had good digestive enzymes tolerability.