Preparation And Identification Of Lipid-lowering Bioactive Peptides From Camel Blood

The death toll from cardiovascular disease accounts for more than 31% of the global death toll,and hyperlipidemia is a dangerous cause of cardiovascular disease,which has become a hot spot of national concern.The pathogenesis of hyperlipidemia is complicated and difficult to prevent.The clinical manifestations are high serum triglyceride and cholesterol levels,and the side effects of drugs used in treatment are large,which often brings severe discomfort to patients.Therefore,there is an urgent need to develop natural-derived bioactive substances to assist in the treatment of hyperlipidemia.In recent years,food-derived bioactive peptides have attracted much attention due to their high safety factor and obvious effects.A large number of studies have shown that camel blood is rich in nutrients and high in total protein content.Many bioactive peptides are encrypted in the primary structure of camel blood protein,which has biological activities such as lowering blood sugar and anti-oxidation.Therefore,this experiment aims to use neutral protease,flavour protease,papain and alkaline protease to hydrolyze camel blood protein,and compare its enzymatic hydrolysis process,the pancreatic lipase inhibition ability,molecular weight distribution and peptide sequence Research in order to obtain highly active bioactive peptides from camel blood for lowering blood lipids.(1)The optimal enzymatic hydrolysis process for different protease hydrolysis camel blood protein is different.Affected by the restriction site,the pancreatic lipase inhibition ability of the enzymatic hydrolysate product is also different.Taking the pancrelipase inhibition rate of the enzymatic hydrolysate product as a judgement index,four proteases with more advantages are selected from the seven proteases.The pancreatic lipase inhibition rate of the enzymatic hydrolysis products obtained under the respective optimal enzymatic hydrolysis process conditions from high to low is: flavour protease hydrolysis product(FW)> papain hydrolysis product(MG)> neutral protease hydrolysis product(ZX)>Alkaline protease digestion product(JX).The optimal enzymatic hydrolysis process of flavor protease is 3% enzyme dosage,enzymatic hydrolysis time 3 h,enzymatic hydrolysis temperature 65 ?,enzymatic hydrolysis pH7.5.SDS-PAGE gel electrophoresis showed that camel blood macromolecular proteins had been effectively enzymatically digested and the enzymatic hydrolysis effect was good.The MALDI-TOF-MS spectrum of FW has the most elution peaks,and there are more small peptides in FW.(2)Peptide sequence analysis showed that there were 144 identical peptides in ZX,FW,MG and JX,and 406,1973,22 and 167 different peptides,respectively.It can be seen that the peptides of the enzymatic hydrolysis products of different proteases are rich in types,mainly from hemoglobin,serum albumin,spectrin and fibrin.Among them,hemoglobin is most decomposed by the four proteases and is the main source of protein decomposition.(3)The pancreatic lipase inhibition rate of fraction 2(FW2)after chromatography of the flavour protease enzymatic hydrolysis product was 51.72%,which was significantly higher than that of fraction 1(FW1)and fraction 3(FW3)(P<0.05).107 peptides with potential biological activity,the molecular weights are mainly concentrated in 400?1400Da.The results of Pepsite 2 show that PFF,FPMF,FPAF and FPHF bind to the active site of pancrelipase most significantly.It was verified by artificial synthesis that the inhibitory ability of a single peptide pancrelipase from strong to weak was PFF>FPMF>FPAF>FPHF.The pancreatic lipase inhibition ability of mixed peptides was the strongest,which was significantly higher than that of 4 single peptides(P<0.05).Compared with BIOPEP,Peptide DB,Swe Pep and EROPMoscow,there is no search record,indicating that new biologically active peptides have been discovered in this experiment.