Preparation method of high-activity fish protein powder
Through the synergistic action of nematose enzymes and microbial fermentation, the problem of insufficient decomposition of proteins in the preparation of traditional fish protein powder is solved, efficient multi-stage degradation of fish proteins and maximum release of nutrients, and the stability and functionality of the product are improved.
Patent Information
- Application Number
- CN202510054698.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-14
- Publication Date
- 2025-04-11
AI Technical Summary
Traditional fish protein powder preparation methods lead to insufficient decomposition of proteins and other macromolecular substances, low content of small peptides and free amino acids in the product, low nutritional value, and prone to deterioration during storage, affecting stability and safety.
The method of synergistic action of bacterase is used to initially hydrolyze deep-sea fish proteins through metalloproteinase and biological coenzyme, and then fermentation of microorganisms such as Bacillus Bacillus Bacillus halophilus, further decompose small-molecular peptides and other organic substances, and combine with drying media to form a nanocapsule structure to protect the stability of the product.
It achieves efficient multi-stage degradation of fish protein, maximizes the release of nutrient active ingredients, improves the quality of the product and the content of functional ingredients, and ensures the stability and safety of the product.
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Figure CN119799829A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of seafood processing, and in particular to a method for preparing high-activity fish protein powder. Background Art
[0002] Fish protein refers to protein extracted from fish, typically obtained from fish (such as deep-sea and freshwater fish) through enzymatic hydrolysis or fermentation. Rich in essential amino acids, fish protein possesses high nutritional value and diverse biological activities. As a high-quality protein source, fish protein can be converted into fish protein powder through enzymatic hydrolysis or microbial fermentation. This product efficiently degrades large fish proteins into small peptides and free amino acids, while retaining and releasing a variety of functional ingredients. It has widespread applications in the food and feed industries, pharmaceuticals, and cosmetics.
[0003] Traditional fish protein powder preparation methods mainly rely on single enzymatic hydrolysis or simple fermentation, resulting in incomplete degradation, failure to fully decompose proteins and other macromolecules, low content of small peptides and free amino acids in the product, and low nutritional value; at the same time, products made by traditional processes are prone to deterioration during storage, affecting their stability and safety; in addition, traditional methods have low efficiency in utilizing the synergistic effects of microorganisms and enzymes, resulting in low levels of functional ingredients such as antioxidants and antimicrobial peptides, resulting in a narrow application range of related products. Summary of the Invention
[0004] In view of the shortcomings of the prior art, the object of the present invention is to provide a method for preparing high-activity fish protein powder.
[0005] A method for preparing high-activity fish protein powder comprises the following steps: (1) Raw material pretreatment: clean the deep-sea fish, remove impurities, crush them using a crusher, transfer the crushed raw materials into the enzymatic hydrolysis tank, add brown sugar and / or molasses, yeast paste and clean water, and mix them evenly; (2) Bacterial enzyme degradation process: metalloproteinases and biological coenzymes are added to the enzymatic hydrolysis tank for preliminary enzymatic hydrolysis; after the preliminary enzymatic hydrolysis, Bacillus velezensis, halophilic Bacillus, Fengda salt-tolerant microorganisms and Microalgae Monosaccharides are introduced for bacterial enzyme degradation to obtain bacterial enzyme degradation liquid; (3) Drying and packaging: The enzyme degradation solution is fully mixed with the chitosan solution, and then sent to a spray drying tower for spray drying to obtain highly active fish protein powder, which is then sealed and packaged and stored in a cool and dry place; The high-activity fish protein powder comprises the following raw materials in parts by weight: 1-20 parts of deep-sea fish, 5-10 parts of brown sugar and / or molasses, 1-5 parts of metalloproteinase, 1-5 parts of biological coenzyme, 1-3 parts of Bacillus velezensis agent, 1-3 parts of halophilic Bacillus, 1-5 parts of yeast extract, 1-10 parts of Fengda salt-tolerant microorganisms, 1-3 parts of microalgae monads, and 1-70 parts of clean water.
[0006] Preferably, the Bacillus Velez, Bacillus halophilus, salt-tolerant microorganisms of Funda and Microalgae Mononas are pre-activated and cultured, the basic formula of the activation culture medium includes brown sugar and / or molasses as a carbon source, yeast extract as a nitrogen source and clean water, the activation culture conditions are pH 6.5-7.0, temperature 30-37° C., and activation time 12-24 hours.
[0007] Preferably, metalloproteinases are added to the activation medium to activate degradation-related genes of the microorganisms in advance.
[0008] Preferably, the pH value of the preliminary enzymatic hydrolysis is 6.5-7.0, the temperature is 30-37° C., and the time is 2-4 hours.
[0009] Preferably, the pH value of the bacterial enzyme degradation is 5.5-7.5, the temperature is 30-50° C., and the time is 24-48 hours.
[0010] Preferably, the amount of chitosan solution used is calculated based on the mass of chitosan, with 10-50 grams of chitosan added to each liter of bacterial enzyme degradation solution.
[0011] Preferably, the chitosan is used as the drying medium, and the drying medium is any one of chitosan, α-cyclodextrin and maltodextrin.
[0012] Preferably, the spray drying temperature is 80-100° C., the feed rate is 1-3 L / h, and the nozzle pressure is 1.5-3.0 bar.
[0013] Preferably, the spray drying is carried out in two steps, first spray drying at 80-100° C. for 20-40 minutes, and then drying at a low temperature below 40° C. under vacuum conditions for 2-4 hours.
[0014] Preferably, before step (3) drying and packaging, an inactivation and termination reaction is also included: the bacterial enzyme degradation solution is heated to 80-90°C for 10-30 minutes to inactivate the enzyme and stop microbial activity, and then quickly cooled to room temperature.
[0015] Description of the raw materials used in the above scheme: 1. Metalloproteases: The presence of metalloproteases makes substrates more easily utilized by microorganisms, thereby improving the efficiency of the entire degradation process. In addition, metalloproteases can activate the degradation-related genes of certain microorganisms, start the metabolic activities of these microorganisms in advance, improve the efficiency of enzymatic hydrolysis and fermentation, accelerate the degradation process, and promote the production of functional ingredients such as antioxidants and antimicrobial peptides, giving the product more health benefits.
[0016] 2. Biocoenzymes: Biocoenzymes such as NADH / NADPH and ATP are essential cofactors in microbial metabolism, enhancing metabolic efficiency and maintaining healthy microbial growth. By participating in various biochemical reactions, they accelerate substrate conversion and ensure the rapid release of nutrients. Biocoenzymes are not only key players in microbial metabolism but also serve as signaling molecules to promote synergy between different microorganisms, forming complex, mutually beneficial symbiotic networks.
[0017] 3. Bacillus Velez agent: As an efficient protein-degrading bacterium, it can secrete a variety of secondary metabolites, such as antimicrobial peptides, plant growth stimulants and other functional metabolites, to improve the quality and function of the final product. Its metabolites not only enhance the health function of the product, but may also have health benefits such as anti-inflammatory and antioxidant. The presence of Bacillus Velez can form a synergistic effect with other microorganisms, optimize the entire degradation process, and improve the stability of the product and the content of functional ingredients.
[0018] 4. Halophilic Bacillus: Halophilic Bacillus has a high degree of environmental adaptability and can remain active under high salinity or other extreme conditions, ensuring the stability and reliability of the entire treatment process. In addition, there is a synergistic effect between Halophilic Bacillus and Bacillus Velez, which improves the overall degradation efficiency.
[0019] 5. Microalgae: also known as single-cell algae or microalgae, is a microorganism capable of photosynthesis. It provides an additional source of energy in the preparation of highly active fish protein powder, supporting the growth and metabolic activities of other microorganisms. The metabolites produced by Microalgae have natural antioxidant effects, protecting other ingredients from oxidative damage and ensuring the stability and activity of functional ingredients.
[0020] The beneficial effects achieved by the present invention are as follows: 1. The present invention uses metalloproteinases and biological coenzymes to preliminarily hydrolyze deep-sea fish proteins through the synergistic effect of bacteria and enzymes, decomposes large molecular proteins into small peptides and free amino acids, improves the efficiency of subsequent microbial treatment, releases trace nutrients originally bound to large molecular proteins, introduces microbial fermentation such as Bacillus velezensis and Bacillus halophilus, further decomposes small molecular peptides and other organic substances, produces more easily absorbed small molecular compounds, and secretes a variety of secondary metabolites, thereby achieving efficient and comprehensive multi-stage degradation of fish protein, maximizing the release of nutritional active ingredients, and helping to improve the quality and function of the final product; finally, the fish protein powder is protected by a drying medium to form a nanocapsule structure, effectively ensuring the stability and safety of the highly active fish protein powder.
[0021] 2. The present invention activates the degradation-related genes of microorganisms through metalloproteinases, so that the microorganisms immediately enter an efficient working state when introduced into the enzymatic hydrolysis tank. Carbon and nitrogen sources are used to provide rich nutrition for the microorganisms, promoting their rapid reproduction and metabolism. Halophilic Bacillus and Fengda salt-tolerant microorganisms are selected to ensure that the entire treatment process is stable and reliable. The microalgae can carry out photosynthesis, providing an additional energy source, and act as a natural antioxidant to protect other components from oxidative damage, thereby enhancing the self-regulation ability and long-term stability of the system.
[0022] 3. The present invention adopts a two-step drying method. First, spray drying is used to quickly remove most of the moisture, so that the drying medium such as chitosan forms a stable nanocapsule structure during the spray drying process, effectively protecting the functional components in the fish protein powder from the influence of the external environment. Then, low-temperature vacuum drying is used to further reduce the moisture content, protect heat-sensitive components, and extend the service life. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 The present invention is a schematic flow chart of the preparation method of the high-activity fish protein powder. DETAILED DESCRIPTION
[0024] The technical solution of the present invention is further described in detail below with reference to specific embodiments.
[0025] In the present invention, unless otherwise specified, all equipment and raw materials can be purchased from the market or are commonly used in the industry. The methods in the following embodiments, unless otherwise specified, are all conventional methods in the art.
[0026] Example 1: A method for preparing highly active fish protein powder, such as Figure 1 As shown, the following steps are included: (1) Raw material pretreatment: 10 kg of deep-sea fish were cleaned, impurities were removed, and crushed using a crusher. The crushed raw materials were transferred to an enzymatic hydrolysis tank, 5 kg of brown sugar, 1.5 kg of yeast paste and 25 liters of clean water were added and mixed evenly; (2) Bacterial enzyme degradation process: prepare activation culture medium, including 2.5 kg brown sugar, 1.5 kg yeast extract and 15 liters clean water, adjust the pH value to 6.8, the temperature to 35 °C, add 1 kg metalloproteinase, inoculate 2 kg of Bacillus velezensis, 1.5 kg of halophilic Bacillus, 5 kg of Fengda salt-tolerant microorganisms and 2 kg of Microalgae Mononas into the activation culture medium, and activate for 12 hours; Add 2 kg of metalloproteinase and 3 kg of biological coenzyme into the enzymatic hydrolysis tank, set the pH value to 6.8, the temperature to 35 ° C, and the time to 3 hours for preliminary enzymatic hydrolysis; The activated microorganisms and their culture medium were introduced into an enzymatic hydrolysis tank for enzyme degradation. The pH value of the enzyme degradation was 6.5, the temperature was 37°C, and the time was 36 hours to obtain an enzyme degradation solution. (3) Inactivation and termination reaction: Heat the enzyme degradation solution to 85°C for 20 minutes and then quickly cool it to room temperature to inactivate the enzyme and stop microbial activity; (4) Drying and packaging: The enzyme degradation solution and chitosan solution were fully mixed, 30 g of chitosan was added to each liter of enzyme degradation solution, and the solution was sent to a spray drying tower for spray drying at a temperature of 90 °C, a feed rate of 2 L / h, and a nozzle pressure of 2.0 bar to obtain highly active fish protein powder. After drying, the solution was sealed and packaged and stored in a cool and dry place.
[0027] Example 2: A method for preparing highly active fish protein powder, such as Figure 1 As shown, the following steps are included: (1) Raw material pretreatment: 15 kg of deep-sea fish were cleaned, impurities were removed, and crushed using a crusher. The crushed raw materials were transferred to an enzymatic hydrolysis tank, and 5 kg of molasses, 2 kg of yeast paste and 35 liters of clean water were added and mixed evenly; (2) Bacterial enzyme degradation process: prepare activation culture medium, including 3 kg of brown sugar, 2 kg of yeast extract and 20 liters of clean water, adjust the pH value to 6.5 and the temperature to 32 °C, add 2 kg of metalloproteinase, inoculate 2.5 kg of Bacillus velezensis, 2.5 kg of Bacillus halophilus, 8 kg of Fengda salt-tolerant microorganisms and 2 kg of Microalgae Mononas into the activation culture medium, and activate for 18 hours; Add 2 kg of metalloproteinase and 3 kg of biological coenzyme into the enzymatic hydrolysis tank, set the pH value to 6.5, the temperature to 32 ° C, and the time to 2 hours for preliminary enzymatic hydrolysis; The activated microorganisms and their culture medium were introduced into an enzymatic hydrolysis tank for enzyme degradation. The pH value of the enzyme degradation was 7.0, the temperature was 45°C, and the time was 30 hours to obtain an enzyme degradation solution. (3) Inactivation and termination reaction: Heat the enzyme degradation solution to 80°C for 30 minutes and then quickly cool it to room temperature to inactivate the enzyme and stop microbial activity; (4) Drying and packaging: The enzyme degradation solution was fully mixed with the α-cyclodextrin solution, 40 g of α-cyclodextrin was added to each liter of enzyme degradation solution, and the solution was sent to a spray drying tower for spray drying. The solution was spray dried at 85 °C for 40 minutes, with a feed rate of 1.5 L / h and a nozzle pressure of 1.8 bar. The solution was then dried under low-temperature vacuum conditions below 40 °C for 2 hours to obtain highly active fish protein powder. After drying, the solution was sealed and packaged and stored in a cool and dry place.
[0028] Example 3: A method for preparing highly active fish protein powder, such as Figure 1 As shown, the following steps are included: (1) Raw material pretreatment: 20 kg of deep-sea fish were cleaned, impurities were removed, and crushed using a crusher. The crushed raw materials were transferred to an enzymatic hydrolysis tank, and 7 kg of brown sugar, 4 kg of yeast paste and 50 liters of clean water were added and mixed evenly; (2) Bacterial enzyme degradation process: prepare activation culture medium, including 3 kg of brown sugar, 1 kg of yeast extract and 20 liters of clean water, adjust the pH value to 7.0 and the temperature to 37 °C, add 2 kg of metalloproteinase, inoculate 3 kg of Bacillus velezensis, 3 kg of Bacillus halophilus, 10 kg of Fengda salt-tolerant microorganisms and 3 kg of Microalgae Mononas into the activation culture medium, and activate for 24 hours; Add 3 kg of metalloproteinase and 5 kg of biological coenzyme into the enzymatic hydrolysis tank, set the pH value to 7.0, the temperature to 37 ° C, and the time to 4 hours for preliminary enzymatic hydrolysis; The activated microorganisms and their culture medium are introduced into an enzymatic hydrolysis tank for enzyme degradation. The pH value of the enzyme degradation is 7.5, the temperature is 50°C, and the time is 24 hours to obtain an enzyme degradation solution. (3) Inactivation and termination reaction: Heat the enzyme degradation solution to 90°C for 10 minutes and quickly cool it to room temperature to inactivate the enzyme and stop microbial activity; (4) Drying and packaging: The enzyme degradation solution and chitosan solution were fully mixed, 50 g of chitosan was added to each liter of enzyme degradation solution, and the solution was sent to a spray drying tower for spray drying at a temperature of 100 °C, a feed rate of 3 L / h, and a nozzle pressure of 3.0 bar to obtain highly active fish protein powder. After drying, the solution was sealed and packaged and stored in a cool and dry place.
[0029] Comparative Example 1: Referring to Example 1, but without using metalloproteinase and biological coenzyme, the preparation method of high-activity fish protein powder includes the following steps: (1) Raw material pretreatment: same as in Example 1; (2) Bacterial enzyme degradation process: prepare activation culture medium, including 2.5 kg brown sugar, 1.5 kg yeast extract and 15 liters clean water, adjust the pH value to 6.8 and the temperature to 35 °C, inoculate 2 kg of Bacillus velezensis, 1.5 kg of halophilic Bacillus, 5 kg of Fengda salt-tolerant microorganisms and 2 kg of Microalgae Mononas into the activation culture medium, and activate for 12 hours; The activated microorganisms and their culture medium were introduced into the enzymatic hydrolysis tank for enzyme degradation. The pH value of the enzyme degradation was 6.5, the temperature was 37°C, and the time was 39 hours (plus the original 3 hours of initial enzymatic hydrolysis) to obtain the enzyme degradation solution. (3) Inactivation and termination reaction: same as in Example 1; (4) Drying and packaging: The same as in Example 1 was used to prepare high-activity fish protein powder.
[0030] Comparative Example 2: Referring to Example 1, but without using Bacillus Velez and Bacillus halophilus, the preparation method of high-activity fish protein powder comprises the following steps: (1) Raw material pretreatment: same as in Example 1; (2) Bacterial enzyme degradation process: prepare activation culture medium, including 2.5 kg brown sugar, 1.5 kg yeast extract and 15 liters clean water, adjust the pH value to 6.8, the temperature to 35 ° C, add 1 kg metalloprotease, inoculate 7 kg Fengda salt-tolerant microorganisms (plus the original Velez Bacillus) and 3.5 kg Microalgae Monosaccharides (plus the original halophilic Bacillus) into the activation culture medium, and the activation time is 12 hours; Add 2 kg of metalloproteinase and 3 kg of biological coenzyme into the enzymatic hydrolysis tank, set the pH value to 6.8, the temperature to 35 ° C, and the time to 3 hours for preliminary enzymatic hydrolysis; The activated microorganisms and their culture medium were introduced into an enzymatic hydrolysis tank for enzyme degradation. The pH value of the enzyme degradation was 6.5, the temperature was 37°C, and the time was 36 hours to obtain an enzyme degradation solution. (3) Inactivation and termination reaction: same as in Example 1; (4) Drying and packaging: same as in Example 1; The high-activity fish protein powder prepared in Examples 1-3 and Comparative Examples 1-2 was subjected to relevant performance tests; 1. Moisture content determination: According to GB / T 6435-2014 "Determination of moisture in feeds", accurately weigh about 2-5 grams of sample and place it in the sample tray of the moisture analyzer. Start the instrument, heat the temperature to 105℃, set the temperature for 30 minutes, record the final reading, and calculate the moisture content percentage.
[0031] 2. Protein content determination: According to GB / T 6432-2018 "Determination of crude protein in feeds", spread the sample evenly on the sample plate to ensure uniform thickness. Use the pre-calibrated model to scan the sample. The analysis software automatically calculates and displays the protein content.
[0032] 3. Determination of total bacterial count: According to GB 4789.2-2016 "National Food Safety Standard - Microbiological Examination of Food - Determination of Total Colony Count", take 1 gram of sample, add 9 ml of sterile saline, mix well, use an ATP swab to dip into the sample solution, wipe it gently, insert it into the detector, start the detector, record the RLU value (relative light unit), and convert it into total bacterial count according to the standard curve.
[0033] 4. pH value determination: According to GB / T 9829-2008 "Determination of pH value in feed", mix the sample with an equal volume of distilled water, stir thoroughly, insert a calibrated pH meter into the sample suspension, and record the pH value after the reading stabilizes.
[0034] 5. Sensory evaluation: In accordance with GB / T 8321.1-2000 "Guidelines for the Rational Use of Pesticides (I)", visual inspection: observe the color, particle size and uniformity of the sample to ensure that there are no foreign matter and lumps; odor inspection: smell the sample for any peculiar or unusual odor to ensure that there is no corruption or other unpleasant odor; color inspection: compare with the standard sample to ensure that the color is consistent and there is no obvious color difference.
[0035] The performance test results are shown in the table below.
[0036] Table 1 Test results of various properties of high-activity fish protein powder
[0037] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Anyone skilled in the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by one of ordinary skill in the art without departing from the spirit and technical principles disclosed herein are intended to be covered by the claims of the present invention.
Claims
1. A method for preparing high-activity fish protein powder, characterized in that: The following steps are involved: (1) Raw material pretreatment: clean the deep-sea fish, remove impurities, crush them using a crusher, transfer the crushed raw materials into an enzymatic hydrolysis tank, add brown sugar and / or molasses, yeast paste and clean water, and mix them evenly; (2) Bacterial enzyme degradation process: metalloproteinases and biological coenzymes are added to the enzymatic hydrolysis tank to perform preliminary enzymatic hydrolysis; after the preliminary enzymatic hydrolysis, Bacillus Velez, Bacillus halophilus, Fengda salt-tolerant microorganisms and Microalgae Mononas are introduced to perform bacterial enzyme degradation to obtain bacterial enzyme degradation liquid; (3) Drying and packaging: The enzyme degradation solution is fully mixed with the chitosan solution, and then sent to a spray drying tower for spray drying to obtain highly active fish protein powder, which is then sealed and packaged and stored in a cool and dry place; The high-activity fish protein powder comprises the following raw materials in parts by weight: 1-20 parts of deep-sea fish, 5-10 parts of brown sugar and / or molasses, 1-5 parts of metalloproteinases, 1-5 parts of biological coenzymes, 1-3 parts of Bacillus Velez, 1-3 parts of halophilic Bacillus, 1-5 parts of yeast extract, 1-10 parts of Fengda salt-tolerant microorganisms, 1-3 parts of microalgae monosaccharides, and 1-70 parts of clean water.
2. The method for preparing a highly active fish protein powder according to claim 1, characterized in that: The Velez Bacillus, halophilic Bacillus, Funda salt-tolerant microorganisms and Microalgae Mononas are pre-activated and cultured, the basic formula of the activation culture medium includes brown sugar and / or molasses as a carbon source, yeast extract as a nitrogen source and clean water, the activation culture conditions are pH 6.5-7.0, temperature 30-37°C, and activation time 12-24 hours.
3. The method for preparing a high-activity fish protein powder according to claim 2, characterized in that: Metalloproteinases were added to the activation medium.
4. The method for preparing a highly active fish protein powder according to claim 1, characterized in that: The pH value of the preliminary enzymatic hydrolysis is 6.5-7.0, the temperature is 30-37° C., and the time is 2-4 hours.
5. The method for preparing a highly active fish protein powder according to claim 1, characterized in that: The pH value of the bacterial enzyme degradation is 5.5-7.5, the temperature is 30-50° C., and the time is 24-48 hours.
6. The method for preparing a high-activity fish protein powder according to claim 1, characterized in that: The amount of chitosan solution is calculated based on the mass of chitosan, and 10-50 grams of chitosan is added to each liter of bacterial enzyme degradation solution.
7. The method for preparing a high-activity fish protein powder according to claim 6, characterized in that: The chitosan is used as a drying medium, and the drying medium is any one of chitosan, α-cyclodextrin and maltodextrin.
8. The method for preparing a high-activity fish protein powder according to claim 1, characterized in that: The spray drying temperature is 80-100° C., the feed rate is 1-3 L / h, and the nozzle pressure is 1.5-3.0 bar.
9. The method for preparing a high-activity fish protein powder according to claim 8, characterized in that: The spray drying is carried out in two steps, firstly spray drying at 80-100° C. for 20-40 minutes, and then drying at a low temperature vacuum condition below 40° C. for 2-4 hours.
10. The method for preparing a high-activity fish protein powder according to claim 1, characterized in that: Before step (3) drying and packaging, the process also includes inactivation and termination of the reaction: heating the enzyme degradation solution to 80-90°C for 10-30 minutes to inactivate the enzyme and stop the microbial activity, and then rapidly cooling the solution to room temperature.