Acid Enzyme Combination Of Bullfrog Skin Collagen Extraction Process Parameters Optimization

This topic is the optional topic, in order to take enhances the use of the bullfrog skin waste, to extract the bullfrog skin collagen. Bullfrog skin deep processing lays theoretical basis, and first applied acid enzyme combination that bullfrog skin extracted successfully the collagen. This experiment mainly research collagen extraction process pretreatment conditions of solutions, the use of acid extract collagen enzyme combination with the optimum technological conditions, use response surface method on four main parameters affecting the extraction process to optimize the obtained optimum extraction program, extract and detect the type of collagen and amino acid composition.First:In extracting collagen protein for bullfrog skin before the pretreatment conditions for process optimization, analysis the pigment and fat that are contained in bullfrog skin of collagen and quality of the influence of guarantee do not affect collagen protein yield, under the premise of bleaching, degreasing are technology processing, through the sensory, single factor experiment, orthogonal experiment, determined the bullfrog skin to extract collagen the best pretreatment process conditions before, and at the same time, the place of different condition available products are compared. Testing to determine the optimum conditions of pretreatment, the liquid materials as than,10%n-butyl alcohol solution as degreasing agents,10℃conditions skim8h. The best decoloring technology for:10℃conditions,0.1mol/L Na OH solution (including1%H2O2solution) decoloring16h. Under the treatment conditions, the bullfrog skin degreasing rate of43.78%.Second:This test of the basic components of bullfrog skin were measured and analyzed by drawing hydroxyproline standard curve, to determine the hydroxyproline content in collagen, and getting bullfrog skin collagen content. Experimental measurement of bullfrog skin of its moisture content was9.71%and19.45%, ash content, fat content of2.35%,42.58%of the total protein, collagen content was28.32%.Third:the type of acid extracted collagen, comparison tests, to determine the type of acid is the best acid. Enzyme amount of collagen extracted, the extraction temperature and extraction liquid ratio, extraction time than four factors were single factor experiment, by the response surface method for the optimization analysis, application software Design Expert after optimization of the optimum technological conditions for:10℃, acetic acid combined with2.5‰pepsin,1:40solid to liquid ratio, extraction24h. Extracted under the condition bullfrog skin collagen content of27.98%.Fourth:Collagen extracted UV spectra measured its maximum absorption peak at222nm at the characteristics of this type I collagen absorption peak match, and can be seen from the spectra of collagen tryptophan low levels of tyrosine and phenylalanine, the collagen of high purity. Collagen extracted from the amino acid composition analysis, the glycine content in about1/3, the total amount of proline and hydroxyproline accounted for23.92%of the total amino acid, methionine, histidine, tyrosine content of very low, these characteristics are consistent with the characteristics of type I collagen. Extracted from bullfrog skin get is I type collagen, and high purity.