Preparation method of fish collagen tripeptide

Abstract

The present application belongs to the technical field of fish collagen preparation, and particularly relates to a preparation method of fish collagen tripeptide. The preparation method comprises raw material pretreatment, collagen peptide crude extraction, deodorization and decolorization, fine filtration, evaporation concentration and drying, so as to obtain fish collagen tripeptide with a molecular weight of 180-500 Dalton. The preparation method is simple in operation and free of organic reagents or inorganic reagents in the production process. The production process is simple, the product quality is high, the quality is excellent, the production energy consumption is saved, the production cost is greatly reduced, the collagen tripeptide content and GPH content of the product are higher than those of existing products on the market, the average molecular weight is small, the product quality is effectively controlled, the product has high safety, and the product has a wide application range.

Description

Technical Field

[0001] This invention belongs to the field of fish collagen preparation technology, specifically relating to a method for preparing fish collagen tripeptide. Background Technology

[0002] Currently, there are some studies on extracting collagen tripeptides from poultry skin and bones. The general process includes steps such as raw material pretreatment, enzymatic hydrolysis, purification, concentration, and spray drying. However, most of these processes use large amounts of organic or inorganic solvents during pretreatment, and employ ultrasound or resin treatment during enzymatic hydrolysis and purification. The production process is cumbersome, the product purity is low, and the production cost is high, thus failing to meet people's needs. Summary of the Invention

[0003] The technical problem to be solved by this invention is to provide a method for preparing fish collagen tripeptide, thereby overcoming at least one of the technical defects existing in the prior art. The adopted technical solution is as follows:

[0004] A method for preparing fish collagen tripeptide includes the following steps:

[0005] (1) Raw material pretreatment: Break the fish skin, soak it in water for 6 to 9 hours, wash it to remove impurities from the fish skin, and obtain clean broken fish skin.

[0006] (2) Extraction: Add the minced fish skin obtained in step (1) to purified water at a weight ratio of 1:4-6, heat to 70-75℃, add high-temperature protease, extract for 4-9 hours, heat to 85-90℃ and then inactivate the enzyme for 15-45 minutes, cool to 40-60℃, add protease A for enzymatic hydrolysis for 4-9 hours, and then inactivate the enzyme to obtain crude collagen peptide extract;

[0007] (3) Deodorization and decolorization: Add 4% to 10% of the weight of fish skin's composite activated carbon to the crude collagen extract obtained in step (2) and stir for 45 to 80 minutes to decolorize and deodorize. Filter to obtain a clear fish collagen solution after deodorization and decolorization.

[0008] (4) Fine filtration: The collagen clear liquid obtained in step (3) is subjected to ceramic membrane, ultrafiltration and nanofiltration to obtain collagen tripeptide stock solution;

[0009] (5) Finished product: The collagen tripeptide stock solution obtained in step (4) is evaporated and concentrated, and then spray-dried to obtain fish collagen tripeptide with a molecular weight of 180-500 Daltons.

[0010] The molecular weight was determined using the method specified in GB / T 22729; the collagen tripeptide content was determined using GB / T16631-2008; and the glycine-proline-hydroxyproline (GPH) content was determined using GB / T 16631-2008.

[0011] Preferably, in step (1), the fish skin is made from one or more of the following: cod, pollock, or haddock, and the water used for washing is tap water.

[0012] Preferably, in step (2), the high-temperature protease is a neutral protease derived from the fermentation of the Geobacillus sp. strain.

[0013] Preferably, in step (2), the total weight of the added high-temperature protease is 0.1% to 0.3% of the weight of the fish skin.

[0014] Preferably, in step (2), the protease A is bromelain refined from pineapple, neutral protease fermented by Bacillus subtilis, alkaline protease fermented by Bacillus licheniformis, and flavor protease fermented by Aspergillus oryzae, with a weight ratio of bromelain: neutral protease: alkaline protease: flavor protease = 1:5~12:5~12:0.2~0.8.

[0015] Preferably, in step (2), the total weight of proteinase A added is 1% to 8% of the weight of the fish skin.

[0016] Preferably, the composite activated carbon is a decolorizing activated carbon and a deodorizing activated carbon, with a weight ratio of 1:0.2 to 1.2 for the decolorizing activated carbon to the deodorizing activated carbon.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] This invention provides a method for preparing fish collagen tripeptides, which is simple to operate and requires no organic or inorganic reagents during production. The production process is simple, the product has high quality and excellent performance, saves energy consumption, and greatly reduces production costs. The collagen tripeptide content and GPH content of the product are higher than those of existing products on the market, the average molecular weight is small, the product quality is effectively controlled, the product has high safety, and the product has a wide range of applications. Attached Figure Description

[0019] Figure 1 This is a flowchart of the preparation method of the present invention. Detailed Implementation

[0020] The accompanying drawings are for illustrative purposes only; it should be understood that the embodiments mentioned below are merely used to explain the present invention, in order to facilitate the description of the present invention and simplify the description, and therefore should not be construed as limiting the present invention.

[0021] The principles and features of the present invention are described below with reference to embodiments. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified in the embodiments, conventional conditions or conditions recommended by the manufacturer should be followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0022] Example 1

[0023] like Figure 1 As shown, a method for preparing fish collagen tripeptide specifically includes the following steps:

[0024] (1) Raw material pretreatment: After crushing the cod skin, soak it in tap water for 6 hours and wash it. The amount of tap water used is 10 times the weight of the fish skin to obtain clean and minced fish skin.

[0025] (2) Extraction: The minced fish skin was added to purified water at a weight ratio of 1:4, heated to 70°C, and high-temperature protease was added. The mixture was extracted for 4 hours, heated to 85°C and then the enzyme was inactivated for 30 minutes. The temperature was lowered to 60°C, and protease A was added for enzymatic hydrolysis for 9 hours. Then the enzyme was inactivated to obtain crude collagen peptide extract.

[0026] The high-temperature protease is a neutral protease derived from the fermentation of *Geobacillus sp.* strain, and the total weight of the enzyme added is 0.3% of the fish skin weight; the protease A is derived from bromelain refined from pineapple, neutral protease fermented by *Bacillus subtilis*, alkaline protease fermented by *Bacillus licheniformis*, and flavor protease fermented by *Aspergillus oryzae*, with a weight ratio of bromelain:neutral protease:alkaline protease:flavor protease = 1:5:5:0.2; the total weight of protease A added is 8% of the fish skin weight;

[0027] (3) Deodorization and decolorization: Add 4% of the weight of fish skin of the crude collagen extract to the extract and stir for 80 minutes to decolorize and deodorize. Filter to obtain clear fish collagen solution after deodorization and decolorization. The composite activated carbon is decolorizing activated carbon and deodorizing activated carbon, with a weight ratio of 1:0.5.

[0028] (4) Fine filtration: The obtained collagen clear liquid is passed through a ceramic membrane, ultrafiltration and nanofiltration to obtain collagen tripeptide stock solution;

[0029] (5) Finished product: The collagen tripeptide stock solution obtained in step (4) is evaporated and concentrated, and then spray-dried to obtain the finished product.

[0030] The molecular weight determination results using the method of GB / T 22729 are shown in Table 1.

[0031] Table 1 shows the molecular weight of the collagen tripeptide prepared in Example 1 of this invention.

[0032]

[0033] Total weight average molecular weight (Mw): 539

[0034] The content of collagen tripeptide was tested according to GB / T 16631-2008; the content of glycine-proline-hydroxyproline (GPH) was tested according to GB / T 16631-2008, and the results are shown in Table 2.

[0035] Table 2 shows the glycine-proline-hydroxyproline (GPH) content in the product prepared in Example 1.

[0036] Test Item Test Method Test Results GPH content (mg / g) GB / T 16631-2008 3.41 Relative content (%) of collagen tripeptide (CTP) GB / T 16631-2008 31.2

[0037] As shown in Tables 1 and 2, the product prepared in Example 1 has a total weight-average molecular weight (Mw) of 539, a GPH content (mg / g) of 3.41, and a relative content (%) of collagen tripeptide (CTP) of 31.2. The product has a low average molecular weight and high collagen tripeptide and GPH content.

[0038] Example 2

[0039] A method for preparing fish collagen tripeptide specifically includes the following steps:

[0040] (1) Raw material pretreatment: After crushing the mixed fish skin of pollock and haddock, soak it in tap water for 9 hours and wash it. The amount of tap water used is 5 times the weight of the fish skin to obtain clean and minced fish skin.

[0041] (2) Extraction: The minced fish skin was added to purified water at a weight ratio of 1:6, heated to 75°C, and high-temperature protease was added. The mixture was extracted for 6 hours, heated to 90°C and then the enzyme was inactivated for 30 minutes. The temperature was lowered to 60°C, and the protease was added for enzymatic hydrolysis for 6 hours. Then the enzyme was inactivated to obtain crude collagen peptide extract.

[0042] The high-temperature protease is a neutral protease derived from the fermentation of *Geobacillus sp.* strain, and the total weight of the enzyme added is 0.2% of the fish skin weight; the protease is derived from bromelain refined from pineapple, neutral protease fermented by *Bacillus subtilis*, alkaline protease fermented by *Bacillus licheniformis*, and flavor protease fermented by *Aspergillus oryzae*, with a weight ratio of bromelain:neutral protease:alkaline protease:flavor protease = 1:10:10:0.4; the total weight of protease A added is 6% of the fish skin weight;

[0043] (3) Deodorization and decolorization: Add 10% of the weight of fish skin to the crude collagen extract and stir for 80 minutes to decolorize and deodorize. Filter to obtain clear fish collagen liquid after deodorization and decolorization.

[0044] (4) Fine filtration: The obtained collagen clear liquid is passed through a ceramic membrane, ultrafiltration and nanofiltration to obtain collagen tripeptide stock solution;

[0045] (5) Finished product: The collagen tripeptide stock solution obtained in step (4) is evaporated and concentrated, and then spray-dried to obtain the finished product.

[0046] The molecular weight determination results using the method of GB / T 22729 are shown in Table 3.

[0047] Table 3 shows the molecular weight of the collagen tripeptide prepared in Example 2 of this invention.

[0048]

[0049] Total weight average molecular weight (Mw): 528

[0050] The content of collagen tripeptide was tested according to GB / T 16631-2008; the content of glycine-proline-hydroxyproline (GPH) was tested according to GB / T 16631-2008, and the results are shown in Table 4.

[0051] Table 4 shows the glycine-proline-hydroxyproline (GPH) content in the product prepared in Example 2.

[0052] Test Item Test Method Test Results GPH content (mg / g) GB / T 16631-2008 3.44 Relative content (%) of collagen tripeptide (CTP) GB / T 16631-2008 32.0

[0053] As shown in Tables 3 and 4, the product prepared in Example 2 has a total weight-average molecular weight (Mw) of 528, a GPH content (mg / g) of 3.44, and a relative content (%) of collagen tripeptide (CTP) of 32.0. The product has a low average molecular weight and high collagen tripeptide and GPH content, both exceeding those of existing products on the market.

[0054] Of course, the above description is not intended to limit the present invention, and the present invention is not limited to the examples given above. Any changes, modifications, additions or substitutions made by those skilled in the art within the scope of the present invention should also fall within the protection scope of the present invention.

Claims

1. A method for preparing fish collagen tripeptide, characterized in that, Includes the following steps: (1) Raw material pretreatment: Break the fish skin, soak it in water for 6 to 9 hours, wash it to remove impurities from the fish skin, and obtain clean broken fish skin; (2) Extraction: Add the minced fish skin obtained in step (1) to purified water at a weight ratio of 1:4-6, heat to 70-75℃, add high-temperature protease, extract for 4-9 hours, heat to 85-90℃ and then inactivate the enzyme for 15-45 minutes, cool to 40-60℃, add protease A for enzymatic hydrolysis for 4-9 hours, and then inactivate the enzyme to obtain crude collagen peptide extract; (3) Deodorization and decolorization: Add 4% to 10% of the weight of fish skin composite activated carbon to the crude collagen peptide extract obtained in step (2) and stir for 45 to 80 minutes to decolorize and deodorize. Filter to obtain clear fish collagen solution after deodorization and decolorization. (4) Fine filtration: The collagen clear solution obtained in step (3) is passed through a ceramic membrane, ultrafiltration and nanofiltration to obtain collagen tripeptide stock solution; (5) Finished product: The collagen tripeptide stock solution obtained in step (4) is evaporated and concentrated, and spray-dried to obtain fish collagen tripeptide with a molecular weight of 180-500 Daltons. The fish skin used is from one or more of the following: cod, pollock, or haddock. The high-temperature protease is a neutral protease fermented from Geobacillus sp. strain. The total weight of the high-temperature protease added is 0.1% to 0.3% of the fish skin weight. The protease A is derived from bromelain refined from pineapple, neutral protease fermented from Bacillus subtilis, alkaline protease fermented from Bacillus licheniformis, and flavor protease fermented from Aspergillus oryzae, with a weight ratio of bromelain: neutral protease: alkaline protease: flavor protease = 1:5 to 12:5 to 12:0.2 to 0.

8. The total weight of protease A added is 1% to 8% of the fish skin weight.

2. The method for preparing a fish collagen tripeptide according to claim 1, characterized in that, In step (1), the water used for cleaning is tap water.

3. The method for preparing a fish collagen tripeptide according to claim 1, characterized in that, The composite activated carbon consists of decolorizing activated carbon and deodorizing activated carbon, with a weight ratio of 1:0.2 to 1.2 for the decolorizing activated carbon to the deodorizing activated carbon.

Images