阅读说明：本技术 一种淡水鱼鱼鳞胶原蛋白肽粉的制备方法 (Preparation method of freshwater fish scale collagen peptide powder ) 是由 尤娟 张皓炎 姜越 尹涛 胡杨 熊善柏 于 2020-05-09 设计创作，主要内容包括：本发明涉及一种淡水鱼鱼鳞胶原蛋白肽粉的制备方法,称取干燥鲈鱼鱼鳞,用蒸馏水冲洗；接着用NaOH溶液进行脱杂蛋白处理,用蒸馏水洗涤至中性；再加入HCl溶液进行脱钙处理,再用蒸馏水洗涤至中性；脱钙后湿鱼鳞与水混合,搅拌,用纱布过滤,得滤液为胶原蛋白原液,冷冻保存备用；将胶原蛋白原液中加入酶量进行酶解反应,结束后置于沸水浴中终止酶解反应；用超滤设备对酶解液进行超滤,得到胶原蛋白肽液；用冷冻干燥机对肽液进行冷冻干燥,然后用气流粉碎机进行处理,得到胶原蛋白肽粉。本发明采用“热水提取、碱性蛋白酶酶解”的工艺,既可以避免其他低价值蛋白肽的掺入,还同时能够确保胶原蛋白肽的均一性及稳定性,延长贮藏时期。(The invention relates to a preparation method of scale collagen peptide powder of freshwater fish, weigh the dry perch fish scale, wash with distilled water; then carrying out deproteinization treatment by using NaOH solution, and washing to be neutral by using distilled water; adding HCl solution for decalcification treatment, and washing with distilled water to neutrality; mixing the decalcified wet fish scales with water, stirring, filtering with gauze to obtain a filtrate as collagen stock solution, and freezing and storing for later use; adding enzyme into the collagen stock solution for enzymolysis reaction, and placing the collagen stock solution in a boiling water bath to terminate the enzymolysis reaction after the enzymolysis reaction is finished; carrying out ultrafiltration on the enzymolysis liquid by using ultrafiltration equipment to obtain collagen peptide liquid; and (3) carrying out freeze drying on the peptide liquid by using a freeze dryer, and then treating by using an airflow pulverizer to obtain collagen peptide powder. The invention adopts the processes of hot water extraction and alkaline protease enzymolysis, thereby not only avoiding the doping of other low-value protein peptides, but also ensuring the uniformity and stability of the collagen peptide and prolonging the storage period.)

1. A preparation method of freshwater fish scale collagen peptide is characterized by comprising the following steps: the method comprises the following steps:

1) weighing dried weever scales, and washing with distilled water;

2) adding NaOH solution to carry out deproteinization treatment, and washing with distilled water to be neutral;

3) adding HCl solution for decalcification treatment, and washing with distilled water to neutrality;

4) mixing the decalcified wet fish scales with water, stirring, filtering with gauze to obtain a filtrate as collagen stock solution, and freezing and storing for later use;

5) adding enzyme into the collagen stock solution for enzymolysis reaction, and placing the collagen stock solution in a boiling water bath to terminate the enzymolysis reaction to obtain collagen peptidase hydrolysate;

6) carrying out ultrafiltration on the enzymolysis liquid by using ultrafiltration equipment to obtain collagen peptide liquid;

7) and (3) carrying out freeze drying on the peptide liquid by using a freeze dryer, and then treating by using an airflow pulverizer to obtain collagen peptide powder.

2. The method of claim 1, wherein: the feed-liquid ratio of the weever scales in the step 2) to the NaOH solution is w/v =1: 10-1: 8, the concentration of the NaOH solution is 0.08-0.12 mol/L, the treatment temperature of the impurity-removed protein is 20-30 ℃, and the treatment time is 4-5 hours.

3. The method of claim 1, wherein: the feed-liquid ratio of the perch scales to the HCL solution in the step 2) is w/v =1: 10-1: 8, the concentration of the HCl solution is 0.4-0.6 mol/L, the impurity removal treatment temperature is 20-30 ℃, and the treatment time is 1.5-2 hours.

4. The method of claim 1, wherein: and 3) mixing the decalcified wet fish scales with water =1: 5-1: 4(w/v), wherein the stirring temperature is 70-75 ℃, and the stirring time is 6-7 hours.

5. The method of claim 1, wherein: the adding amount of the enzyme in the step 4) is 0.50-1.00% of the mass of the protein stock solution, the enzyme is alkaline protease, the enzymolysis pH is 9-10, the enzymolysis temperature is 35-40 ℃, and the enzymolysis time is 30-40 min.

6. The method of claim 1, wherein: in the step 1), before the washing with distilled water, yeast is added for fermentation treatment, the mass ratio of the weever scales to the yeast is 100 (0.5-1.5), and the fermentation treatment is carried out for 3-4 hours at the temperature of 25-35 ℃.

Technical Field

The invention belongs to the field of freshwater fish processing, and particularly relates to a preparation method of freshwater fish scale collagen peptide powder.

Background

The fish scales contain a large amount of protein and mineral substances, wherein organic matters account for 41-55 percent. A large amount of byproducts generated in the processing process of the special freshwater fish are high in nutritive value and are increased along with the increase of the total processing amount of aquatic products in China, but a large amount of fish processing byproducts are processed into animal feed or are directly discarded, so that not only is great resource waste caused, but also the environment is seriously polluted. Therefore, the research on the processing by-products of the freshwater fish is developed, so that the comprehensive benefits of the processing and utilization of aquatic products in China and the economic added value of the freshwater fish can be improved, and the environmental pollution can be avoided. At present, more researches and reports on fish scale collagen active peptide exist, but in raw material selection, the tilapia, which is a tropical fish, is mostly used as a carrier to prepare the collagen peptide, and some enterprises also use carp fishes such as grass carp and the like as carriers to prepare the collagen peptide; meanwhile, in the extraction and enzymolysis process, the existing method also has different degrees of influence on the stability and uniformity of the collagen peptide product, and the production process of 'complex enzyme extraction and complex enzyme enzymolysis' is mostly adopted, but the production process can mix proteins which are contained in fish scales and have low nutritive value for human bodies, such as sclerostin, and the like, into the collagen, so that the relative nutritive value of the collagen peptide product is reduced; in addition, some enterprises adopt a production process of acid extraction and pepsin enzymolysis, which can avoid the problems, but the production process takes a long time and reduces the enterprise efficiency for production.

Disclosure of Invention

The invention provides a preparation method of freshwater fish scale collagen peptide powder, which takes characteristic freshwater fish processing by-product scales as raw materials, takes weever scales as research carriers, and comparatively screens collagen endonucleases suitable for scale oriented enzymolysis to produce the high-value high-homogeneity collagen peptide powder.

The technical scheme adopted by the invention is as follows:

a preparation method of freshwater fish scale collagen peptide powder comprises the following steps:

1) weighing dried weever scales, and washing with distilled water;

2) adding NaOH solution to carry out deproteinization treatment, and washing with distilled water to be neutral;

3) adding HCl solution for decalcification treatment, and washing with distilled water to neutrality;

4) mixing the decalcified wet fish scales with water, stirring, and filtering with gauze to obtain filtrate as collagen stock solution;

5) adding enzyme into the collagen stock solution for enzymolysis reaction, and placing the collagen stock solution in a boiling water bath to terminate the enzymolysis reaction after the enzymolysis reaction is finished; obtaining collagen peptidase hydrolyzed solution.

6) Carrying out ultrafiltration on the enzymolysis liquid by using ultrafiltration equipment to obtain collagen peptide liquid;

7) and (3) carrying out freeze drying on the peptide liquid by using a freeze dryer, and then treating by using an airflow pulverizer to obtain collagen peptide powder.

Preferably, the feed-liquid ratio of the weever scales in the step 2) to the NaOH solution is 1: 10-1: 8, the concentration of the NaOH solution is 0.08-0.12 mol/L, the treatment temperature for removing the impurity protein is 20-30 ℃, and the treatment time is 4-5 hours.

Preferably, the feed-liquid ratio of the weever scales in the step 3) to the NaOH solution is 1: 10-1: 8, the concentration of the HCl solution is 0.4-0.6 mol/L, the impurity removal treatment temperature is 20-30 ℃, and the treatment time is 1.5-2 hours.

Preferably, the wet fish scales and water after decalcification in the step 4) are mixed at a stirring temperature of 70-75 ℃ for 6-7 hours, wherein the ratio of water to water is 1: 5-1: 4 (w/v).

Preferably, in the step 5), the adding amount of the enzyme is 0.50-1.00% of the mass of the protein stock solution, the enzyme is alkaline protease, the enzymolysis pH is 9-10, the enzymolysis temperature is 35-40 ℃, and the enzymolysis time is 30-40 min.

Preferably, in the step 1), before the washing with distilled water, yeast is added for fermentation treatment, the mass ratio of the weever scales to the yeast is 100 (0.5-1.5), and the fermentation treatment is carried out for 3-4 hours at the temperature of 25-35 ℃.

The invention has the following beneficial effects:

1. the method takes the fish scales of the special freshwater fish processing by-products as raw materials, takes the fish scales of weever as research carriers, comparatively screens out the collagen incision enzyme suitable for the directional enzymolysis of the fish scales, optimizes the enzymolysis technology, evaluates the practicability and effectiveness of the fish scales by a popular sensory evaluation method, removes the fishy smell contained in the raw materials, improves the purity, uniformity and absorption efficiency of the enzymolysis collagen short peptide, produces the high-value collagen high-uniformity collagen peptide powder, explores the storage performance of the collagen high-value collagen peptide powder, and realizes the high-value utilization of the special freshwater fish processing by-products.

2. The invention adopts the processes of hot water extraction and alkaline protease enzymolysis, can avoid the doping of other low-value protein peptides, greatly shorten the production time, ensure the uniformity and stability of the collagen peptide, prolong the storage period and bring greater benefits for enterprises.

Drawings

The invention is further illustrated by the following figures and examples.

FIG. 1 hydroxyproline standard curve;

FIG. 2 different enzyme treatment particle size distributions;

FIG. 3 shows particle size distribution of alkaline protease at different treatment times;

FIG. 4 shows the molecular weight distribution of peptide fragments of fish scale collagen peptide prepared by different enzymes;

FIG. 5 shows the effect of different enzymes on SDS-PAGE bands of a collagen stock solution of a fish scale of weever;

FIG. 6 shows the change of total thiol content with prolonged storage time at different humidity;

FIG. 7 comparison of the change in endogenous fluorescence intensity at different storage stages at different humidity;

FIG. 8 sensory evaluation scores with prolonged storage time;

Detailed Description

The invention is further described with reference to the following figures and detailed description.