Study On Extraction, Nature And Peptide Activity Of Collagen From The Yellow River Carp Fish-scale

In the machining process of the Yellow River carp, fish scales were mostlydiscarded as waste or processing of fish feed, which contains rich collagen is not verygood use. Collagen has a natural, low toxicity, biodegradability, low immune activity,moisturizing and nutrition and is widely used in the food industry, cosmetics,biomedical materials field. In this paper, collagen is extracted from carp fish scales,and the extraction process is optimized. The research of the nature and the activity ofcollagen peptide provides a theoretical basis for the development and utilization ofcarp fish scale collagen. The contents and results are as follows:1. Determination of collagen content measured by hydroxyproline standard curvemethod, the test measured the standard curve: y=0.0652x+0.0028, R²=0.9995, toprove that good linear relationship of concentration and absorbance. The bestconditions for scales demineralization process optimization are: hydrochloric acidconcentration of1.0mol/L, liquid ratio of8:1, the removal time of150min, theremoval rate of92.4%.2. Scale collagen extraction process optimization results: optimum conditions ofacid extraction are the concentration of acetic acid solution of0.54mol/L, theextraction time of42h, extract feed ratio of10:1, under the conditions of thisparameter, the Yellow River carp ichthyosis collagen extraction rate of the theoreticalprediction value is15.75%, the actual measured extraction rate is16.89%. Optimalparameters of enzyme extraction are pepsin solution concentration of13.50mg/mL, theextraction time of12h, extract feed ratio of10:1, under the conditions of thisparameter, the theoretical prediction of the Yellow River carp fish scale collagenextraction rate is15.06%, and the actual measured extraction rate is16.75%.3. UV scans showed carp fish scales PSC and ASC has a strong absorption at232nm wavelength, in line with the characteristics of type I collagen. Infrared spectrum scanning analysis shows in3074.71cm^-1,2933.16cm^-1,2869.42cm^-1, near the placeof weak absorption is collagen characteristic absorption peak. SDS-PAGE gelelectrophoresis showed that PSC and ASC has three chains, three chains, two α chain（α₁, α₂） and a β chain （two α chain crosslinked body）, this and related literaturereported the characteristics of type I collagen consistent. Water absorption and water-holding capacity of the test can directly see the extracted collagen gel with good waterabsorption, poor water absorption of commercially available gelatin; commerciallyavailable gelatin scale collagen extracted water retention contrast we can see: fishscale collagen certain amount of moisture can be maintained within the storage period,the effect is better than the commercially available gelatin.4. The research results of collagen peptide activity: the hydrolysis of neutralprotease AS1398is the strongest, and the degree of hydrolysis is71.84%,corresponding enzymatic product inhibition of ACE activity rate effect is the best, theinhibition rate of51.43%. Enzymatic hydrolysis process select AS1398neutralprotease optimal enzyme. Conducted tests on several important factors ACE collagenpeptide extraction process can be seen: the extraction temperature of45℃, pH7.5,enzyme and substrate quality volume ratio of1.5%, and liquid to solid ratio of20:1foroptimum extraction conditions, under this condition, the degree of hydrolysis is75.9%,ACE inhibition rate was55.8%.Components I, II, III and IV were higher than ACEinhibition hydrolysis without ultrafiltration treatment liquid, component IV （<2000u）ACE inhibitory highest rate of82.9%, while the rate of hydrolysis of ACE inhibitorydope only72.4%.