Continuous fluid extraction method of collagen peptide

Abstract

The invention provides a continuous fluid extraction method of collagen peptide, and belongs to the technical field of natural protein peptide extraction. The method comprises the following steps: carrying out deliming and / or degreasing treatment on animal raw materials, homogenizing the obtained material, heating, transferring the homogenized material to an enzymolysis tank, adding protease into the enzymolysis tank, adjusting the pH value of the system, carrying out enzymolysis treatment and filtration in sequence, heating the filtered material to a decoloration temperature in the conveying process, and carrying out refined filtration after the decoloration is finished, so as to obtain the collagen peptide. According to the method provided by the invention, the use efficiency of equipment is improved, and the equipment investment is saved; the uniformity of the auxiliary materials is improved in an auxiliary material feeding mode; meanwhile, the collagen peptide powder can be obtained by shortening the whole process time to 4-8 hours, so that the pollution risk is greatly reduced, the product quality is improved, and the purpose of saving the production energy consumption is achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of natural protein peptide extraction, and particularly relates to a continuous fluid extraction method for collagen peptide. Background Art

[0002] Bioactive peptides are a general term for different peptides formed by 25 natural amino acids in proteins with different compositions and arrangements, ranging from dipeptides to complex linear and cyclic structures. They are multifunctional compounds derived from proteins. Bioactive peptides have various functions in human metabolism and physiological regulation, are easily digested and absorbed, and have effects such as promoting immunity, hormone regulation, antibacterial, antiviral, lowering blood pressure, and lowering blood lipids. They have extremely high food safety and are currently the most popular research topic in the international food industry and functional factors with great development prospects.

[0003] The sources of bioactive peptides are mainly proteins obtained from organisms, and most are the reuse of processing by-products. For example, active peptides are processed with fish skin, fish scales, cowhide, soybean meal, etc. as raw materials; some are also produced for the need of promoting absorption, such as collagen peptide, sea cucumber peptide, etc. The extraction methods of bioactive peptides include water extraction, acid extraction, alkali extraction, and enzyme extraction. The extraction of active peptides by enzymatic hydrolysis has been relatively mature technically. The principle is to remove impurities from the target protein, and through certain parameters such as the ratio of material to water, temperature, enzyme amount, pH, and time, cut the macromolecular protein into small pieces and dissolve it in water, and remove the insoluble matter from the material after the reaction. Further operations such as decolorization, deodorization, and refinement are carried out on the enzymatic hydrolysate, and then concentration, sterilization, and drying are carried out. The above processes of cleaning, crushing, enzymatic hydrolysis, filtration, decolorization, clarification, concentration, re-clarification, sterilization, and drying all require reaction kettles to hold the solution. Each batch of materials enters the next process after the reaction in a single process is completed. The materials have temperature rise and fall times, waiting times for the current batch of materials to be transported into the tank, and output times after the reaction ends at each step. It often takes 24 - 48 hours from feeding to drying out the powder, but the core processing time for the materials (enzymatic hydrolysis, decolorization) is only 2 - 3 hours, and the rest of the time is static waiting for feeding and discharging or temperature rise and fall. The disadvantages of this traditional scheme are particularly obvious: 1. The number of required equipment is large and the investment is high; 2. The longer the material is online, the greater the influence of heat treatment on color and smell; 4. The longer the online time, the greater the risk of bacterial contamination, directly resulting in deterioration of product quality. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a continuous fluid extraction method for collagen peptide, which greatly reduces the material production time and ensures product quality.

[0005] The present invention provides a continuous fluid extraction method for collagen peptide, comprising the following steps: Perform deashing and / or degreasing treatment on animal raw materials to obtain pretreated materials; Homogenize the pre-treated material to obtain a homogenate; During the transportation of the homogenate to the enzymolysis tank, heat it up to the enzymolysis temperature, and at the same time add protease and adjust the pH value of the system to the enzymolysis tank. After the transfer is completed, directly carry out enzymolysis treatment, inactivate the enzyme, and obtain an enzymolysis product; Carry out primary filtration on the enzymolysis product, heat it up to the decolorization temperature during the transportation of the obtained primary filtrate material, and after decolorization and fine filtration, obtain collagen peptides.

[0006] Preferably, the animal raw material includes at least one of the following: animal skin, animal bones, animal tendons, animal sinews, fish scales, and animal meat.

[0007] Preferably, the animal bones or fish scales include acid deashing and alkali degreasing; The animal skin, animal tendons, animal sinews, or animal meat includes alkali degreasing.

[0008] Preferably, the particle size of the material in the homogenate is 0.01 - 0.1 mm.

[0009] Preferably, the protease includes at least one of the following: neutral protease, alkaline protease, papain, and trypsin.

[0010] Preferably, the primary filtration or fine filtration is completed by using one of the following instruments: plate and frame filter press, candle filter, and centrifuge.

[0011] Preferably, the initial filtration pressure of the plate and frame filter press is 0.01 MPa, maintained for 15 min, and then the filtration pressure is increased to 0.1 - 0.15 MPa.

[0012] Preferably, the decolorization is completed by using activated carbon; the addition amount of the activated carbon is 5% - 30% of the solid content in the primary filtrate material; During the decolorization process, the activated carbon is added during the transportation of the primary filtrate material.

[0013] Preferably, after the fine filtration, a fine filtrate is obtained, and it further includes concentrating, sterilizing, and drying the fine filtrate.

[0014] Preferably, the concentration method includes using a nanofiltration membrane, falling film type double-effect or triple-effect concentration equipment, or scraper concentration equipment to complete; The sterilization is completed by using pulsed intense light sterilization method; The drying includes spray drying; the inlet air temperature of the spray drying is 170 - 200 °C, the outlet air temperature is 70 - 100 °C, and the atomizer frequency is 30 - 50 HZ.

[0015] A continuous fluid extraction method for collagen peptides of the present invention comprises the following steps: performing deashing and / or degreasing treatment on animal raw materials to obtain pretreated materials; homogenizing the pretreated materials to obtain a homogenate; heating the homogenate to an enzymatic hydrolysis temperature during the conveying process to an enzymatic hydrolysis tank, and simultaneously adding protease and adjusting the system pH value to the enzymatic hydrolysis tank, directly performing enzymatic hydrolysis treatment after the transfer is completed, inactivating the enzyme to obtain an enzymatic hydrolysis product; performing primary filtration on the enzymatic hydrolysis product, heating to a decolorization temperature during the conveying process of the obtained primary filtered material, and performing decolorization and fine filtration to obtain collagen peptides. After the animal raw materials are pretreated by deashing and / or degreasing in the present invention, it is beneficial to remove fat and ash in the animal raw materials, reduce impurity components, and lay a foundation for subsequent enzymatic hydrolysis. At the same time, in the present invention, homogenization is performed after pretreating the animal raw materials and before enzymatic hydrolysis, reducing the volume of the raw materials, which is beneficial to shortening the enzymatic hydrolysis time and improving the uniformity of the molecular weight distribution of enzymatic hydrolysis. In addition, in the present invention, the processes of adjusting the enzymatic hydrolysis temperature, pH value, and adding protease are synchronized during the transfer process of the enzymatic hydrolysis environment materials, and the feeding method is adopted, omitting the time for adjusting the enzymatic hydrolysis conditions after feeding, greatly shortening the overall duration of the preparation process, achieving the purpose of continuous extraction, thereby reducing the probability of product browning and avoiding product deterioration and bacterial contamination during long-term extraction. Experiments show that in the enzymatic hydrolysis process, the traditional method is used to adjust the pretreated materials to suitable conditions in the enzymatic hydrolysis tank and then add enzyme solution for enzymatic hydrolysis. Since the enzymatic hydrolysis process involves heating, adjusting the pH value, and feeding large-volume materials, it takes a long time, resulting in a greatly reduced enzymatic hydrolysis efficiency. However, in the present invention, by synchronizing the processes of adjusting the pH value, enzyme solution, and heating measures during the material conveying process, the material transfer time and the time for heating and adjusting the pH after the transfer ends can be saved, greatly shortening the enzymatic hydrolysis process time and effectively improving the enzymatic hydrolysis efficiency, thereby ensuring the quality of the enzymatic hydrolysis product. It can be seen that the method provided by the present invention can greatly improve the product quality by shortening the extraction cycle, ensuring that the prepared collagen peptides have good color, smell, and are not contaminated or deteriorated.

[0016] At the same time, the method provided by the present invention improves the use efficiency of equipment and saves equipment investment; the material addition method is changed from the traditional per-tank addition to the feeding method, improving the uniformity of auxiliary materials; the entire process time is shortened to 4 - 8 hours to prepare collagen peptide powder; the risk of process pollution is sharply reduced, and the product quality is better; production energy consumption is saved.

[0017] Furthermore, the present invention specifically limits the particle size of the materials in the homogenate to 0.01 - 0.1 mm, which is convenient for conveying and heating, the material particle size is small, the enzymatic hydrolysis uniformity is high, and energy consumption is saved; in addition, the present invention also specifically limits the sterilization to adopt pulsed intense light sterilization technology, which has no hindrance at low temperature, and the low temperature ensures that the product has the best color, smell, and taste, does not undergo thermal reactions, and the unobstructed sterilization improves the material passing efficiency. Description of the Drawings

[0018] Figure 1 This is the equipment composition diagram of the continuous fluid extraction method of collagen peptide for the implementation of the present invention. Note: 1: Raw material pretreatment unit, 2: Feeding unit, 3: Meat grinder, 4: Colloid mill, 5: Conveying pipeline, 6: Enzymolysis tank, 7: Plate and frame filter press, 8: Turnover tank, 9: Concentration tower, 10: Drying tower. Detailed implementation mode

[0019] The present invention provides a continuous fluid extraction method for collagen peptide, which includes the following steps: Perform deashing and / or degreasing treatment on animal raw materials to obtain pretreated materials; Homogenize the pretreated materials to obtain a homogenate; During the conveying process of transferring the homogenate to the enzymolysis tank, heat it to the enzymolysis temperature, add protease and adjust the pH value of the system at the same time. After the transfer is completed, directly perform enzymolysis treatment and inactivate the enzyme to obtain an enzymolysis product; Successively filter, decolorize and finely filter the enzymolysis product to obtain collagen peptide.

[0020] The present invention performs deashing and / or degreasing treatment on animal raw materials to obtain pretreated materials.

[0021] In the present invention, the animal raw materials preferably include at least one of the following: animal skin, animal bones, animal tendons, animal sinews, fish scales and animal meat. The species of the animals preferably include cattle, sheep, horses, donkeys and mules. The fish include marine fish and / or freshwater fish. In the embodiments of the present invention, the continuous fluid extraction method of collagen peptide is illustrated by taking bovine bone, tilapia skin and cod skin as examples respectively.

[0022] In the present invention, the animal bones or fish scales preferably include acid deashing and alkali degreasing. The animal skin, animal tendons, animal sinews or animal meat preferably include alkali degreasing. The acid for acid deashing preferably includes organic acid and / or inorganic acid; such as hydrochloric acid, sulfuric acid, phosphoric acid, citric acid. When performing acid deashing, the volume concentration of the acid is preferably 1% - 5%, and can be 3.5%; the treatment time of the acid is preferably 12 - 48h, and can be 24h. The alkali in the alkali degreasing includes sodium hydroxide or potassium hydroxide. When performing alkali degreasing, the mass concentration of the alkali is preferably 0.1% - 1%, and can be 0.5%; the treatment time of the alkali is preferably 12 - 72h, and can be 48h. The materials after treatment are washed with water until the pH value is within the range of 7 - 9.

[0023] After obtaining the pretreated materials, the present invention homogenizes the pretreated materials to obtain a homogenate.

[0024] In the present invention, the homogenization is to gradually perform crushing treatment on the pretreated materials to form a homogenate with uniform particles. The methods of the crushing treatment successively include a meat grinder and a colloid mill.

[0025] In the present invention, the particle size of the materials in the homogenate preferably includes 0.01 - 0.1 mm, more preferably 0.02 - 0.08 mm, may also be 0.04 - 0.07 mm, or may be 0.05 - 0.06 mm. The material particles in the homogenate are uniform.

[0026] After obtaining the homogenate, in the present invention, the homogenate is heated to the enzymatic hydrolysis temperature during the transportation process to the enzymatic hydrolysis tank, while adding protease and adjusting the pH value of the system. After the transfer is completed, enzymatic hydrolysis treatment is directly carried out, and the enzyme is inactivated to obtain the enzymatic hydrolysis product.

[0027] In the present invention, the protease preferably includes at least one of the following: neutral protease, alkaline protease, papain, and trypsin. It can be a combination of neutral protease and alkaline protease, a combination of neutral protease and papain, a combination of papain and trypsin, a combination of alkaline protease and papain, a combination of papain and trypsin, and a combination of alkaline protease and trypsin. It can also be a combination of neutral protease, alkaline protease, and papain, a combination of neutral protease, alkaline protease, and trypsin, a combination of alkaline protease, papain, and trypsin, or can be a combination of neutral protease, alkaline protease, papain, and trypsin. When the protease contains more than two kinds, each protease is compounded according to equal enzyme activity. The addition amount of the protease is 1500 - 2000 U / g 蛋白 , or may be 1500 U / g 蛋白 . The method of adjusting the pH value of the system is preferably adjusted according to the optimal pH value of each protease. For example, the optimal pH value of neutral protease is 6.5 - 8; the optimal pH value of alkaline protease is 9 - 11; the optimal pH value of papain is 5 - 7; the optimal pH value of trypsin is 8 - 9. When the protease is a combination, it is preferred that each protease performs enzymatic hydrolysis step by step. The temperature of the enzymatic hydrolysis is preferably 50 - 60 °C, and can be 55 °C.

[0028] In the present invention, the method of inactivating the enzyme preferably includes high-temperature enzyme inactivation. The temperature of the high-temperature enzyme inactivation is preferably 90 - 98 °C, and can be 95 °C. The time of the high-temperature enzyme inactivation is preferably 5 - 15 min, and can be 10 min. The purpose of inactivating the enzyme is to inactivate the protease and avoid excessive enzymatic hydrolysis.

[0029] After obtaining the enzymatic hydrolysis product, in the present invention, the enzymatic hydrolysis product is successively subjected to primary filtration, decolorization, and fine filtration to obtain collagen peptides.

[0030] In the present invention, the primary filtration or fine filtration is preferably completed by using one of the following instruments: plate and frame filter press, candle filter, and centrifuge. The initial filtration pressure of the plate and frame filter press is preferably 0.01 MPa and is maintained for 15 min, and then the filtration pressure is preferably increased to 0.1 - 0.15 MPa. The centrifugal speed of the centrifuge is preferably 3000 - 15000 r / min and can be 4000 - 10000 r / min.

[0031] In the present invention, the decolorization is preferably completed by using activated carbon. The added mass of the activated carbon is preferably 5% - 30% of the solids in the primary filtered material and can be 10%. The solution temperature during decolorization is preferably 50 - 90°C and can be 70°C. The purpose of decolorization is to remove unnecessary impurities in the filtrate.

[0032] In the present invention, after fine filtration, a fine filtrate is obtained, and preferably, concentration, sterilization, and drying of the fine filtrate are further included.

[0033] In the present invention, the concentration method preferably includes using a nanofiltration membrane, falling film type double-effect or triple-effect concentration equipment, or scraping concentration equipment to complete. The concentration is preferably concentrated to 2 - 5 times the original volume. The sterilization preferably includes using a pulsed intense light sterilization method to complete. The parameters of the pulsed intense light sterilization method are for a pulsed sterilizer with 30 lamp tubes, the peak power of a single lamp tube is greater than 1.5 megawatts, and the spectral range is 190 nm to 1200 nm. The drying preferably includes spray drying; the inlet air temperature of the spray drying is preferably 170 - 200°C and can be 180 - 190°C; the outlet air temperature is preferably 70 - 100°C and can be 80°C; the atomizer frequency is preferably 30 - 50 Hz and can be 40 Hz.

[0034] In the present invention, after drying, packaging and quality control are also preferably included. The packaging is preferably completed under aseptic conditions, and the packaging requirements are firm sealing, straight sealing line, no false sealing, accurate weighing during bagging, clear labeling, and clean and tidy packaging materials. The quality control includes metal detection, average molecular weight detection, sensory detection, color detection, and other index detections. The qualified requirement for metal detection is that no metal foreign objects corresponding to the specifications of the metal detection test block are detected. The average molecular weight detection includes detection according to the GB31645 standard. The color detection is preferably to prepare a 10% aqueous solution with the prepared collagen peptide and measure the absorbance at 420 nm. The other index detections include detections according to the GB31645 standard.

[0035] In the present invention, preferably, a device for implementing the continuous fluid extraction method of collagen peptide is also provided, as shown in Figure 1, including a raw material pretreatment unit, a feeding unit, a meat grinder, a colloid mill, a conveying pipeline, an enzymatic hydrolysis tank, a plate and frame filter, a turnover tank, a concentration tower and a drying tower that are connected in sequence through pipelines. The raw material pretreatment unit is used for the pretreatment of animal raw materials. The raw material pretreatment unit is preferably an animal raw material soaking pool. The present invention has no special restrictions on the shape, specifications and materials of the soaking pool. The feeding unit is used to transport the pretreated animal raw materials into the meat grinder. The feeding unit is preferably a conveyor belt. The meat grinder and the colloid mill are used to crush animal raw materials to form a homogeneous slurry. The conveying pipeline is used to transport the original slurry to the enzymatic hydrolysis tank, and the side wall of the conveying pipeline is also connected to an acid-base tank, a protease tank and a heating device through pipelines, which is used to synchronously add protease, pH value adjusting reagent and heating of materials during transportation, shortening the time for adjusting pH value, temperature and adding protease after feeding, so as to shorten the fluid extraction cycle. The plate and frame filter is used for primary filtration and fine filtration. A heating jacket is also arranged at the filtration outlet of the plate and frame filter, which is used to heat the primary filtered material to the decolorization temperature. The turnover tank is used for temporary storage and decolorization of the primary filtered material to meet the decolorization requirements. The concentration tower is used for concentration. The drying tower is used for vacuum drying. When the device of the present invention is used to realize continuous extraction of collagen peptides, the degree of continuous extraction is greatly improved. At the same time, since the feeding, pH value adjustment, protease addition and enzymatic hydrolysis temperature are synchronized, the entire cycle of the extraction process is greatly shortened, the utilization rate of each device is improved, the product yield is increased within a certain time, and at the same time, problems such as bacterial contamination, color browning and deterioration are avoided, and the product quality is greatly improved.

[0036] The following is a detailed description of a continuous fluid extraction method for collagen peptides provided by the present invention in conjunction with embodiments, but they cannot be understood as a limitation on the protection scope of the present invention.

[0037] Example 1 A continuous extraction method for collagen derived from tilapia skin 1. Pretreatment: Using fresh tilapia skin as raw material, adding 0.5% sodium hydroxide based on the mass of the raw material, adding water according to a material-water ratio of 1:3, and treating for 24 h for degreasing. After completion, wash the raw material until the pH value is 8 and set aside.

[0038] 2. Crushing (homogenization): Mix the washed raw material with water according to a ratio of dry protein (the amount of water removed from the raw material) to water of 1:5, transfer the material liquid to an emulsification tank, and process it through emulsification equipment (colloid mill, homogenizer) to make the particle size of the slurry 0.01 cm, and there is no particle feeling when touching by hand. Before transportation, adjust the parameters of the heating jacket to make the temperature of the slurry at the outlet of the heating jacket 55 °C and prepare to start transportation; Among them, the dry protein content = protein content / (1 - moisture content).

[0039] 3. Enzymatic hydrolysis: After the transportation starts, add neutral protease from Bacillus subtilis into the transportation pipeline (the filtering port with a heating jacket) at a ratio of 1500 U / g 蛋白 , and conduct an enzymatic hydrolysis reaction at 50°C for 60 min (the reaction time starts from the material transportation time, and the reaction time is longer than the transportation time. After the transportation ends, the material stays in the enzymatic hydrolysis tank to reach the enzymatic hydrolysis time). After the reaction ends, raise the temperature to 90°C and keep it warm for 15 min to complete enzyme inactivation, obtaining the enzymatically hydrolyzed product after enzyme inactivation.

[0040] 4. Primary filtration: Clarify the enzymatically hydrolyzed product using a plate and frame filter press, and adjust the pressure of the plate and frame filter press to 0.1 - 0.2 mpa; the pressure difference between the inlet and outlet shall not exceed 0.1 mpa, obtaining the primary filtrate.

[0041] 5. Decolorization: Adjust the temperature of the heating jacket of the transportation pipeline at the filter outlet to raise the temperature of the primary filtered material to 60°C after passing through the heating jacket. Add activated carbon for decolorization and acid to the pipeline at the back end of the heating jacket. The added mass of activated carbon is 5% of the solid content in the primary filtered material. Start timing for 30 min from the addition of carbon (the filtration time is less than 30 min). After decolorization ends, prepare for fine filtration.

[0042] 6. Fine filtration: Select a plate and frame filter press for fine filtration. At the initial stage of filtration, the pressure is 0.01 mpa. After maintaining it for 15 min, gradually increase the filtration pressure to between 0.1 mpa and 0.15 mpa, obtaining the fine filtrate.

[0043] 7. Concentration: Concentrate the fine filtrate using a nanofiltration membrane, and the concentration of the discharged material at the outlet is 30% (mass percentage of solid content).

[0044] 8. Sterilization: Sterilize the concentrated liquid. The specific conditions are a pulsed sterilizer with 30 lamp tubes, the peak power of a single lamp tube is greater than 1.5 megawatts, and the spectral range is from 190 nm to 1200 nm, obtaining the sterilized material.

[0045] 9. Drying: Spray-dry the sterilized material, control the inlet air temperature of the spray drying tower to be 170°C, the outlet air temperature to be 90°C, and the atomizer frequency to be 23 HZ - 25 HZ, obtaining the collagen peptide powder from tilapia skin.

[0046] 10. Inner packaging: Package under aseptic conditions to obtain the finished product; 12. Detection of other product indicators 1) Average molecular weight: Detect the molecular weight distribution according to the detection method in the GB31645 standard.

[0047] 2) Sensory evaluation: Develop a sensory evaluation form (see Table 1) and have 5 people evaluate.

[0048] 3) Color: Determined by absorbance value. Prepare a 10% (mass percentage) solution and measure the absorbance value at 420 nm.

[0049] 4) Other physical and chemical indicators: Comply with the provisions of GB31645 standard.

[0050] Table 1 Sensory Evaluation Table

[0051] Results: 1. The detection results of the average molecular weight of the product are shown in Table 2. The proportion of small molecular peptides below 1000 Da is more than 80%.

[0052] Table 2 Average Molecular Weight

[0053] 2. Sensory evaluation 5 people evaluated that the taste had no abnormal smell and bitterness, and the smell had a very slight collagen characteristic flavor without abnormal smell.

[0054] 3. Color: OD 420nm is 0.06.

[0055] 4. Other physical and chemical indicators: Comply with the provisions of GB 31645 standard. At the same time, no metal foreign objects are detected.

[0056] Example 2 A continuous extraction method for collagen from fresh cowhide 1. Pretreatment: Using fresh cowhide as raw material, add 1% (by mass of the raw material) of sodium hydroxide, add water according to the ratio of material to water of 1:4, and treat for 48 h for degreasing. After completion, wash the fresh cowhide with water until the pH value is 8 - 9.

[0057] 2. Crushing (homogenization): The washed raw materials are put into the emulsifying tank according to the ratio of dry protein to water of 1:10, and treated by a colloid mill to obtain a slurry with a particle diameter of 0.02 cm for the materials in the slurry, and there is no particle feeling when touched by hand. Before transportation, adjust the parameters of the heating jacket so that the temperature of the slurry at the outlet of the heating jacket is 50 °C, and prepare to start transportation; 3. Enzymatic hydrolysis: After the transportation starts, add alkaline protease from Bacillus subtilis into the transportation pipeline (after the outlet of the heating jacket) at a ratio of 1000 U / g 蛋白 for enzymatic hydrolysis reaction at 55 °C for 30 min (the reaction time starts from the material transportation time, the reaction time is longer than the transportation time, and the material stays in the temporary storage tank after the transportation ends to reach the enzymatic hydrolysis time). After completion, raise the temperature to 95 °C and keep it warm for 15 min to inactivate the enzyme, obtaining an enzyme-inactivated enzymatic hydrolysis product.

[0058] 4. Primary filtration: Use a centrifuge for clarification, centrifuge at a speed of 4000 r / min, and separate the supernatant as the primary filtrate.

[0059] 5. Decolorization: Adjust the heating jacket of the conveying pipeline at the outlet of the filter to make the temperature of the material reach 70°C after passing through the heating jacket. Add activated carbon for decolorization and acid to the pipeline at the rear end of the heating jacket. The amount of activated carbon is 8% of the mass of solids in the solution (mass of solids = volume of solution × concentration of solution). Start timing 60 minutes after adding the carbon (filtration time is less than 60 minutes). After decolorization is completed, prepare for fine filtration.

[0060] 6. Fine Filtration: Select a plate and frame filter press for fine filtration. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining for 15 minutes, gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain fine filtrate; 7. Concentration: The concentration method uses falling film type triple effect concentration, and the outlet material concentration is 45% (mass percentage of solids).

[0061] 8. Sterilization: The sterilization method is pulsed intense light sterilization technology. The specific parameters are a pulsed sterilizer with 30 lamp tubes, the peak power of a single lamp tube is greater than 1.5 MW, and the spectral range is from 190 nm to 1200 nm.

[0062] 9. Spray Drying: The drying method uses spray drying. Control the inlet air temperature of the spray drying tower to be 170°C and the outlet air temperature to be 70°C, and the atomizer frequency to be 23 HZ - 25 HZ to obtain collagen peptides from fresh cowhide.

[0063] 10. Inner Packaging: Packaging is carried out under aseptic conditions; Packaging requirements: The seal is firm, the sealing line is straight, there is no false seal, the bagging weighing is accurate, the label is clear, and the packaging material is clean and tidy.

[0064] 11. Metal Detection: No metal foreign objects corresponding to the specifications of the metal detection test block shall be detected.

[0065] 12. The detection methods for other product indicators are the same as those recorded in Example 1.

[0066] Results 1. The results of the average molecular weight of the product are shown in Table 3. The proportion of small molecule peptides below 1000 Da is approximately 90%.

[0067] Table 3 Average Molecular Weight

[0068] 2. Sensory Evaluation 5 people evaluated that the taste has no strange smell and bitterness, and the smell has a very slight characteristic flavor of collagen and no strange smell.

[0069] 3. Color: OD 420nm is 0.08.

[0070] 4. Other physical and chemical indicators: comply with the provisions of GB 31645 standard. Meanwhile, no metal foreign matters are detected.

[0071] Example 3 A continuous extraction method of collagen from fresh fish scales 1. Pretreatment: Using fresh fish scales as raw materials, add 1% hydrochloric acid by the mass of the raw materials for deashing, add water according to the material-water ratio of 1:2.5, wash off the acid solution after 24h, then add 1% sodium hydroxide by the mass of the raw materials, add water according to the material-water ratio of 1:2.5, and treat for 48h for degreasing. After that, wash the fresh fish scales with drinking water until the pH value is 8 - 9.

[0072] 2. Crushing (homogenization): Put the washed raw materials into the emulsification tank according to the material-water ratio controlled by dry protein of 1:8, and process through a series of emulsification equipment. The size of the material is 0.1 cm, and there is no granular feeling when touched by hand. Adjust the parameters of the heating jacket before transportation to make the temperature of the slurry at the outlet of the heating jacket 60 °C, and prepare to start transportation; 3. Enzymatic hydrolysis: After the transportation starts, add papain into the transportation pipeline (after the outlet of the heating jacket) at a ratio of 2000 U / g 蛋白 for reaction for 90 min (the reaction time is counted from the start time of material transportation, the reaction time is longer than the transportation time, and the material stays in the temporary storage tank after transportation ends to reach the enzymatic hydrolysis time). After that, raise the temperature to 95 °C and keep it warm for 10 min to inactivate the enzyme, obtaining the inactivated enzymatic hydrolysis product.

[0073] 4. Primary filtration: The inactivated enzymatic hydrolysis product is clarified using a centrifuge, and centrifuged at a speed of 4000 r / min for a certain time, and the supernatant is collected.

[0074] 5. Decolorization: Adjust the heating jacket of the transportation pipeline at the outlet of the filter to make the temperature of the material reach 65 °C after passing through the heating jacket. Add activated carbon and acid for decolorization to the pipeline at the back end of the heating jacket. The amount of activated carbon is 10% of the solid content in the solution. Start timing from the addition of carbon for 60 min (the filtration time is less than 60 min). After decolorization, prepare for fine filtration.

[0075] 6. Fine filtration: The decolorized liquid is finely filtered using a plate and frame filter press. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining for 15 min, gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate; 7. Concentration: Concentrate the fine filtrate using a nanofiltration membrane with a pore size of 150 Da to make the concentration of the liquid reach 25% (mass percentage of solid content), obtaining the concentrated liquid.

[0076] 8. Sterilization: Sterilize the concentrated liquid using pulsed intense light sterilization technology. The specific parameters are a pulsed sterilizer with 30 lamps, the peak power of a single lamp is greater than 1.5 megawatts, and the spectral range is from 190 nm to 1200 nm, obtaining the sterilized concentrated liquid.

[0077] 9. Drying: The sterilized concentrated solution is spray-dried. The inlet air temperature of the spray drying tower is controlled at 200 °C, the outlet air temperature is 100 °C, and the atomizer frequency is 25 HZ to obtain the collagen powder derived from fresh fish scales.

[0078] 10. Inner packaging: Packaging is carried out under sterile conditions; packaging requirements: the seal is firm, the sealing line is straight, there is no false seal, the weighing during bagging is accurate, the labeling is clear, and the packaging material is clean and tidy.

[0079] 11. Metal detection: No metal foreign objects corresponding to the specifications of the metal detection test block shall be detected.

[0080] 12. The determination methods of other product indicators are the same as those recorded in Example 1.

[0081] Results 1. The results of the average molecular weight of the product are shown in Table 4. The proportion of small molecular peptides below 1000 Da is about 80%.

[0082] Table 4 Average molecular weight of the product

[0083] 2. Sensory evaluation 5 people evaluated that the taste had no strange smell or bitterness, the smell had a very slight characteristic flavor of collagen, and there was no strange smell.

[0084] 3. Color: OD 420nm is 0.07.

[0085] 4. Other physical and chemical indicators: comply with the provisions of GB 31645 standard. At the same time, no metal foreign objects are detected.

[0086] Comparative Example 1 A continuous extraction method of collagen derived from fresh fish scales, without adopting the preparation process of adding materials and enzymes simultaneously 1. Pretreatment: Using fresh fish scales as raw materials, add 1% hydrochloric acid based on the mass of the raw materials for deashing, add water according to the material-water ratio of 1:2.5, wash off the acid solution after 24 h, then add 1% sodium hydroxide based on the mass of the raw materials, add water according to the material-water ratio of 1:2.5, and treat for 48 h for defatting. After completion, wash the fresh fish scales with domestic drinking water until the pH value is 8-9.

[0087] 2. Enzymolysis: The washed fish scales are fed into the enzymolysis tank, heated to 60 °C, and papain is added in a ratio of 2000 U / g 蛋白 for enzymolysis for 90 min. After completion, heat up to 95 °C and keep warm for 10 min to inactivate the enzyme, obtaining the inactivated enzymolysis product.

[0088] 3. Primary filtration: The enzyme-hydrolyzed product after enzyme inactivation is clarified by a centrifuge. Centrifuge at a speed of 4000 r / min for a certain time, and collect the supernatant and transfer it to the decolorization tank.

[0089] 4. Decolorization: After all the slurry in one tank is centrifuged, heat the decolorizing solution to 65°C, add activated carbon to the supernatant. The amount of activated carbon is 10% of the substrate mass, and keep it warm at 65°C for 60 min. After decolorization, prepare for fine filtration.

[0090] 5. Fine filtration: Fine filter the decolorized slurry with a plate and frame filter press. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining it for 15 min, gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate; 6. Concentration: Concentrate the fine filtrate using a nanofiltration membrane with a pore size of 150D molecular weight to make the slurry concentration reach 25% to obtain the concentrated solution.

[0091] 7. Sterilization: Sterilize the concentrated solution using pulsed intense light sterilization technology. The specific parameters are a pulsed sterilizer with 30 lamp tubes, the peak power of a single lamp tube is greater than 1.5 megawatts, and the spectral range is from 190 nm to 1200 nm to obtain the sterilized concentrated solution.

[0092] 8. Drying: Spray-dry the sterilized concentrated solution, control the inlet air temperature of the spray drying tower to be 200°C, the outlet air temperature to be 100°C, and the atomizer frequency to be 25 HZ to obtain the collagen powder from fresh fish scales.

[0093] 9. Inner packaging: Package under aseptic conditions; Packaging requirements: The seal is firm, the sealing line is straight, there is no false seal, the bagging and weighing are accurate, the label is clear, and the packaging material is clean and tidy.

[0094] 10. Metal detection: No metal foreign matter corresponding to the specification of the metal detection test block shall be detected.

[0095] 11. The determination methods of other product indicators are the same as those recorded in Example 1.

[0096] Results 1. The average molecular weight results of the product are shown in Table 5. The proportion of small molecular peptides below 1000 Da is about 43%.

[0097] Table 5 Average molecular weight of the product

[0098] 2. Sensory evaluation Evaluated by 5 people: All felt that the taste was significantly bitter and there was a relatively obvious fishy smell.

[0099] 3. Color: OD 420nm is 0.157.

[0100] 4. Other physical and chemical indicators: Comply with the provisions of GB 31645 standard. At the same time, no metal foreign matters are detected.

[0101] Comparative Example 2 A continuous extraction method of collagen derived from tilapia skin 1. Pretreatment: Using fresh tilapia skin as raw material, adding 0.5% sodium hydroxide based on the mass of the raw material, adding water according to the ratio of material to water of 1:3, and treating for 24 h for defatting. After completion, wash the raw material until the pH value is 8 and set aside.

[0102] 2. Enzymatic hydrolysis: Feed the washed fish skin into the enzymatic hydrolysis tank, heat up to 50 °C, and add neutral protease derived from Bacillus subtilis at a ratio of 1500 U / g 蛋白 . Enzymatically hydrolyze for 60 min, and after completion, heat up to 95 °C and keep warm for 10 min to inactivate the enzyme, obtaining the inactivated enzymatic hydrolysis product.

[0103] 3. Primary filtration: Clarify the enzymatic hydrolysis product using a plate and frame filter press, adjust the pressure of the plate and frame filter press to 0.1 - 0.2 mpa; the pressure difference between the inlet and outlet shall not exceed 0.1 mpa, obtaining the primary filtrate.

[0104] 4. Decolorization: After all the liquid in one tank is centrifuged, heat up the decolorizing liquid to 60 °C, add activated carbon to the supernatant, and the amount of activated carbon is 5% of the substrate mass, and keep warm at 60 °C for 30 min. After decolorization, prepare for fine filtration.

[0105] 5. Fine filtration: Select the plate and frame filter press for fine filtration. For the initial filtration of the plate and frame filter, the pressure is 0.01 mpa. After maintaining for 15 min, gradually increase the filtration pressure to between 0.1 mpa and 0.15 mpa, obtaining the fine filtrate.

[0106] 6. Concentration: Concentrate the fine filtrate using a nanofiltration membrane, and the concentration of the outlet material is 30% (mass percentage of solids).

[0107] 7. Sterilization: Sterilize the concentrated liquid using pulsed intense light sterilization technology. The specific conditions are a pulsed sterilizer with 30 lamp tubes, the peak power of a single lamp tube is greater than 1.5 megawatts, and the spectral range is from 190 nm to 1200 nm, obtaining the sterilized material.

[0108] 8. Drying: Spray-dry the sterilized material, control the inlet air temperature of the spray drying tower to be 170 °C, the outlet air temperature to be 90 °C, and the atomizer frequency to be 23 HZ - 25 HZ, obtaining the collagen peptide powder derived from tilapia skin.

[0109] 9. Inner packaging: Package under aseptic conditions to obtain the finished product; Results: 1. The detection results of the average molecular weight of the product are shown in Table 6. The proportion of small molecular peptides below 1000 Da is about 61%.

[0110] Table 6 Average molecular weight

[0111] 2. Sensory evaluation Evaluation by 5 persons: All felt that the taste was significantly bitter and the smell had an obvious fishy smell.

[0112] 3. Color: OD 420nm It is 0.132.

[0113] 4. Other physical and chemical indicators: Conform to the provisions of GB 31645 standard. At the same time, no metal foreign objects are detected.

[0114] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A continuous fluid extraction method for collagen peptides, characterized in that, It includes the following steps: Debone and / or degrease the animal raw materials to obtain pretreated materials; Homogenize the pretreated materials to obtain a homogenate; During the transportation of the homogenate to the enzymolysis tank, heat it up to the enzymolysis temperature, and at the same time add protease to the enzymolysis tank and adjust the system pH value. After the transfer is completed, directly carry out enzymolysis treatment, inactivate the enzyme, and obtain an enzymolysis product; Carry out primary filtration on the enzymolysis product, heat it up to the decolorization temperature during the transportation of the obtained primary filtered materials, and after decolorization and fine filtration, obtain collagen peptides.

2. The continuous fluid extraction method of collagen peptide according to claim 1, characterized in that, The animal raw materials include at least one of the following: animal skins, animal bones, animal tendons, animal sinews, fish scales, and animal meat.

3. The continuous fluid extraction method of collagen peptide according to claim 2, characterized in that, The animal bones or fish scales include acid deboning and alkali degreasing; The animal skins, animal tendons, animal sinews, or animal meat include alkali degreasing.

4. The continuous fluid extraction method of collagen peptide according to claim 1, characterized in that, The particle size of the materials in the homogenate is 0.01 - 0.1 mm.

5. The continuous fluid extraction method of collagen peptide according to claim 1, wherein The protease includes at least one of the following: neutral protease, alkaline protease, papain, and trypsin.

6. The continuous fluid extraction method of collagen peptide according to claim 1, wherein The primary filtration or fine filtration is completed by using one of the following instruments: plate and frame filter press, candle filter, and centrifuge.

7. The continuous fluid extraction method of collagen peptide according to claim 6, characterized in that The initial filtration pressure of the plate and frame filter press is 0.01 MPa, maintained for 15 min, and then the filtration pressure is increased to 0.1 - 0.15 MPa.

8. The continuous fluid extraction method of collagen peptide according to claim 1, characterized in that The decolorization is completed by using activated carbon; The addition amount of the activated carbon is 5% - 30% of the solid content in the primary filtered materials; The activated carbon is added during the transportation of the primary filtered materials.

9. The continuous fluid extraction method of collagen peptide according to any one of claims 1 to 8, characterized in that, After fine filtration, a fine filtrate is obtained, and it further includes concentrating, sterilizing, and drying the fine filtrate.

10. The continuous fluid extraction method of collagen peptide according to claim 9, characterized in that, The method of concentration includes using nanofiltration membrane, falling film type double-effect or triple-effect concentration equipment, or scraper concentration equipment to complete; The sterilization is completed by using pulsed intense light sterilization method; The drying includes vacuum drying; the inlet air temperature of the vacuum drying is 170 - 200 °C, the outlet air temperature is 70 - 100 °C, and the atomizer frequency is 30 - 50 Hz.

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