Separation And Characterization Of Protein Components In Bovine Tendon Acellular Matrix Materials

In this paper,the matrix of fresh bovine tendon decellularization is used as the raw material to study the preparation process,physical and chemical properties and functional properties of bovine tendon collagen.First,the basic ingredients of bovine tendon decellularized matrix material were measured;second,bovine tendon decellularized matrix material was used to extract collagen;and finally,the collagen samples obtained after separation and purification were studied for relevant physical and chemical properties and functionality.The test results are as follows:1.The moisture content of bovine tendon decellularized matrix material is 2.328%,the ash content is 2.3%,the fat content is 11.4%,and the protein content is 32.65%.2.The best process conditions for the extraction of bovine tendon decellularized matrix collagen by acid method are:acetic acid concentration 0.05 mol/L,material-liquid ratio 1:35,pH 2.5,extraction temperature 50℃,extraction time 48 h,under this condition The highest extraction rate was 40.12%.The order of the influence of each test factor on the collagen extraction rate was:pH>material-liquid ratio>extraction time>extraction temperature.3.The best process conditions for extraction of bovine tendon decellularized matrix collagen by ultrasound-assisted acid method are:ultrasonic time 17.6584 min,ultrasonic power 177.11 W,material-liquid ratio 1:30,pH 2.48,extraction temperature 51.58℃,extraction time 36 h Under this condition,the extraction rate is the highest,59.67%.The order of the influence of each test factor on the extraction rate of collagen is:ultrasonic power>ultrasonic time>extraction temperature>pH.Compared with acid extraction,the extraction rate of ultrasound-assisted acid extraction of collagen has been significantly improved by about 19.55%,and the extraction time has been greatly shortened by about 12 h.4.Using ultraviolet absorption spectroscopy and SDS-PAGE gel electrophoresis to confirm that the acid method and ultrasound-assisted acid method are collagen products with high purity and complete structure.Among them,the water absorption index of acid-extracted collagen is 12.91±0.12,the oil absorption index is about 28.02±0.22;the ultrasound-assisted acid-extracted collagen has a water absorption index of about 13.08±0.15,and the oil absorption index is about 28.22±0.19.5.Explore the influence of pH,NaCl concentration and temperature on the solubility of collagen.The results show that when pH=2,NaCl concentration is 0.2 mol/L and temperature is 40℃,it is the highest dissolution rate of each influencing factor.The highest solubility values??were 53.94%,61.41%,and 61.85%,and the highest solubility of ultrasound-assisted acid extraction collagen were 55.27%,63.54%,and 66.96%,respectively.In the range of values,the solubility of ultrasound-assisted acid-extracted collagen is slightly higher than that of acid method,and there is no significant difference between the two.6.Explore the effect of pH and NaCl concentration on the foaming and foaming stability,emulsifying and emulsifying stability of collagen.The results show that the best conditions for foaming are pH=12 and NaCl concentration is 0.2 mol/L At this time,the foamability of acid-extracted collagen is 48.04%and 44.54%,and the ultrasound-assisted acid-extracted collagen is 49.18%and 45.13%;the best conditions for foaming stability are pH=8 and NaCl concentration of 0.1 mol/At L,the foaming stability of acid-extracted collagen was 96.51%and 92.98%,and ultrasound-assisted acid-extracted collagen was 97.66%and 93.11%;the best conditions for emulsification were pH=2 and NaCl concentration of 0.4 mol/L At this time,the emulsification of acid-extracted collagen is 55.72%and 50.04%,and the ultrasound-assisted acid-extracted collagen is 56.15%and 50.26%;the best conditions for emulsion stability are pH=8 and NaCl concentration of 0.1 mol/L.The emulsion stability of acid-extracted collagen was 96.07%and 97.79%,and the ultrasound-assisted acid-extracted collagen was 96.08%and 97.81%.7.The DPPH,ABTS and FRAP methods were used to investigate the in vitro antioxidant activity of bovine tendon collagen.The results showed that both collagens have a certain antioxidant activity,and the antioxidant capacity increases with the increase of the concentration.The anti-oxidation effect of ultrasonic method is slightly better than that of acid method,but there is no significant difference.The IC₅₀0 of acid-and ultrasound-assisted acid-extracted collagen for DPPH clearance is 12.13 mg/mL and 12.03 mg/mL,respectively;the VC equivalents contained in acid-extracted and ultrasound-assisted acid-extracted collagen for ABTS clearance Approximately 0.039 umol/mg and 0.040 umol/mg;acid and ultrasound-assisted acid extraction of collagen to the FRAP clearance rate of the equivalent VC content per mg of collagen were 7.414 ug and 8.088 ug,respectively.8.The electron microscope was used to analyze the microstructure of collagen.The electron microscope conformation of acid-extracted collagen and ultrasound-assisted acid-extracted collagen are loose and porous honeycomb-like sheet-like structures.The ultrasound method is more hierarchical and the arrangement is more loose.