Computer Simulation Of Proteolysis Of Protein To Produce Antihypertensive Peptides And Production Of ACE Inhibitory Peptides From Hydrolysates Of Defatted Wheat Germ Protein In Continuous Member Reactor

Owning to the high safety and no sides effects,to produce the ACE（angiotensin I-converting enzyme） inhibitory peptides from food source is evoking great interest in treating and preventing high blood pressure.However,in the present study to produce ACE inbibitory peptides,chose Enzyme is a tedious,time-consuming and blind.So it is key important to improve it,bioinformatics can do it.In this study,an ACE inhibitory peptide bioinformatics’ analysis system was built with MySQL and PHP language,which containing information of ACE inhibitory peptides and proteases collected from published documents and bioinformatics’ web,functions of search ACE inhibitory peptides in the selected protein sequence,theoretically digested protein sequence,gastrointestinal proteolysis in silico and related result analysis were developed.Rubisco protein from alfalfa and casein were selected to be analyzed in this system,the result shows:to the dates in the system,the large subunit of Rubisco protein contains 217 ACE inhibitory peptides,while small subunit contains 49 ACE inhibitory peptides.Chymotrypsin C,Proteinase K,Thermolysin and Bomelain are good proteases to produce ACE inhibitory peptides from Rubisco,and Chymotrypsin C was the best choice.The ACE inhibitory activity of hydrolysate of Chymotrypsin C,Proteinase K and Bomelain are better than that hydrolysate from gastrointestinal proteolysis of Rubisco in silico,so it is valuable to produce ACE inhibitory peptides with protease which are not be from intestinal tract.To casein,αs1-Casein contained 65 kinds ACE inhibitory peptides and amount to 90 peptides;αs2-Casein contained 56 kinds and amount to 78 peptides;β-Casein has 87 kinds and up to 114 peptides;κ-Casein has 55 kinds and up to 68 peptides.Gastrointestinal digestion can only release a few ACE inhibitory peptides.So it is useful to produce ACE inhibitory peptides from the enzyme which are not come from gastrointestinal tract.To test prediction sequences of IC₅₀ of hydrolysate of casein with differe enzyme in experiment,the prediction results shows good accordance with the experiment results, so it is valueable to help chose enzyme in an bioinformatic way.There are several prombles in production of ACE inhibitory peptides from Defatted Wheat Germ Protein（DWPG）,such as low conversion rate aod enzyme can not be reused.In order to increase its production and overcome these prombels, production of ACEI peptides from hydrolysates of DWPG in continuous membrance reactor were studied.In this study,the suitable enzyme and membrance to product ACE inhibitory peptides from hydrolysates of DWPG were chose,the influences of hydrolysis time,enzyme concentration（[E]/[S]）,hydrolysis temperature,pH,peptides flux on the conversion rate of DWPG without substrates feeding were studied.Base on these,the mathematic model was established to describe the process of continuous hydrolysis of DWPG with the stability analysis.The production ability and operation stability to product the ACE inhibitory peptide from DWGP in continuous membrane reactor with the optimized substrate concentration from the mathematic model and the stability of product（DWPG＜5000） were invested.（1） Owing to the sequence of deflated wheat grem protein were unkown,it can not to choose enzyme in the assist analytic system.So the quantitative structure-activity relationship（QSAR） of ACE inhibitory peptides was built to get the structure information of ACE inhibitory peptides（R=0.928）,base on the structure information,Acalase and Neutrase were chosen as the candidate enzymes.Through the analysis of DH and ACE inhibitory rate of hydrolysate of different enzymes, Acalase was chosen to produce ACE inhibitory peptides from hydrolysates of DWPG. The DH of wheat germ protein with Acalase were fitted with the process of concentration and time,using L-B method to deal with ditferent substrate concentrations and initial reaction rate,kinetic constants Km=2.234gl^-1 and Vmax= 0.1022 gl^-lmin^-1 were obtained.（2） Several membranes with molecular weight cut-offs（MWCFs） of 10KDa、5kDa and1kDa were used to filter the hydrolysate.The 5kDa MWCF membrane was selected by assessing the IC₅₀ and recovery rate of.infiltration liquid of different MWCFs.The constructed memrance reactor can intercept of substrance and enzyme. Enzyme can be proected by substance.（3） The influences of hydrolysis time,enzyme concentration（[E]/[S]）,hydrolysis temperature,pH,peptides flux on the conversion rate of DWPG without substrates feeding were studied.Base on these,the mathematic model was established to describe the process of continuous hydrolysis of DWPG with the stability analysis. The production ability and operation stability to product the ACE inhibitory peptide from DWGP in continuous membrane reactor was invested with the optimized substrate concentration from the mathematic model,Km=8.1633 g l^-1:Vmax=0.7895 g l^-1min^-1.The results show that taking advantage of the membrane reactor,the protein convection rate and utilization efficiency of enzyme can be increased 26.30%and 4.625 times,respectively.During the process,both the concentration and the ACE inhibitory rate of the product were keep stability.Therefore,membrane reactor provided an efficient way to prepare active peptides.（4） Although the IC₅₀ of HDWGP＜5000 increase from 0.45 mg/ml to 0.86mg/ml, it also means good ACE inhibitory activity.In addition,HDWGP＜5000 has good high temperature stability and good tolerance of the acid-base.