Preparation, Structural Characterization And In Vitro Efficacy Of Yanbian Cattle Bone Collagen Peptide

Bovine bone collagen peptide is currently the most common exogenous bone collagen peptide.The protein content in bovine bone from different varieties and different parts is different.Studies have shown that the protein content in the ribs of Yanbian Yellow Cattle is higher than that in other parts of the bovine bone,but there are few studies on it and it has not been fully used.In order to achieve high-value by-products of bovine bone,Yanbian Yellow Cattle rib was used as the raw material in this paper to prepare bovine bone collagen peptide by enzyme method,analyze its amino acid composition,and resolve its structure by chromatography,electrophoresis,and electron microscopy,to study the in vitro efficacy of bovine bone collagen peptide,which would provide technical support for the comprehensive utilization of Yanbian Yellow Cattle rib.The specific results are as follows:(1)Preparation of bovine bone collagen peptide.Using Yanbian Yellow Cattle rib as raw material,hydrolysis degree as index,the optimal enzyme for collagen peptide preparation from bovine bone meal was screened out as trypsin from five enzymes.On the basis of single factor experiment,the optimum enzymolysis conditions were obtained by response surface methodology:enzyme-substrate ratio 7.28:100,pH8,enzymolysis time 4 h,enzymolysis temperature 37?;Under these conditions,the degree of hydrolysis reached 12.26%.(2)Structural characterization of bovine bone collagen peptide.The amino acid composition and content of bovine bone collagen peptide were analyzed,The total amino acid(TAA)content is 332.72 mg/g,of which Gly 43.97 mg/g is the most,which is the classical amino acid composition of collagen;the UV spectrum results show that the bovine bone collagen peptide has the strongest absorption peak at 227 nm;the infrared spectrum results show that the bovine bone collagen peptide has a characteristic absorption peak at 3325-1 cm;the SDS-PAGE results show that the bovine bone collagen peptide has a characteristic absorption peak at 150 kDa ? Belt,at about 130 kDa ?1??2 two electrophoresis bands;Scanning electron microscopy showed that bovine bone collagen showed a relatively smooth lamellar structure;There was no strong X-ray diffraction peak in the X-ray diffraction pattern of bovine bone collagen peptide.(3)Effects of bovine bone collagen peptide in vitro.The scavenging abilities of the bovine bone collagen peptide prepared by enzymolysis for different free radicals are concentration-dependent,and the scavenging abilities are enhanced with the increase of concentration,so the bovine bone collagen peptide has good in vitro antioxidant effect;The IC50 values for DPPH·,ABTS+,OH,and superoxide radicals scavenging were 9.46 mg/mL,1.55 mg/mL,4.35 mg/mL,and 7.67 mg/mL,respectively.Bovine bone collagen peptide had a certain inhibitory effect on ?-glucosidase but the inhibitory ability did not increase when the concentration reached a certain level.It had a promoting effect on alpha-amylase;The binding rate of bovine bone collagen peptide cholate was less than 20%and did not change with the increase of concentration.Bovine bone collagen peptide showed a good inhibitory effect on ACE activity with the IC50 value of 4.52 mg/mL.To sum up,the collagen peptide of Yanbian Yellow Cattle bone prepared by trypsin hydrolysis belongs to type I collagen peptide,which has good antioxidant,?-glucosidase inhibition and ACE activity in vitro.And it has certain cholate binding ability.Therefore,the study on extracting collagen peptide from ribs of Yanbian Yellow Cattle can provide reference for screening and preparing products with antioxidant,hypoglycemic and hypotensive functions.