CN109097428B

CN109097428B - Preparation method of fish protein peptide

Info

Publication number: CN109097428B
Application number: CN201811014730.1A
Authority: CN
Inventors: 叶晨
Original assignee: Ningbo Haizhiyuan Biotechnology Co ltd
Current assignee: Ningbo Haizhiyuan Biotechnology Co ltd
Priority date: 2018-08-31
Filing date: 2018-08-31
Publication date: 2021-04-06
Anticipated expiration: 2038-08-31
Also published as: CN109097428A

Abstract

The invention relates to the technical field of biology, and particularly discloses a preparation method of a fish protein peptide, which comprises a raw material preparation step, wherein the raw material comprises marine low-value small trash fish and aquatic product leftovers; crushing raw materials; fish paste enzymolysis: transferring the fish paste in the crushing step into an enzymolysis tank, and adding water into the enzymolysis tank, wherein the mass ratio of the water to the fish paste is 1: 1, mixing water and fish paste to form a mixture A, wherein the enzymolysis temperature is 50-70 ℃, the pH value during enzymolysis is 7.0, and the enzymolysis time is 2-6 h; enzyme deactivation; preparing small peptide liquid; spray drying the enzymolysis liquid; and (3) microbial fermentation: mixing Aspergillus oryzae into the solid in the small peptide liquid preparation step to form a mixture C, wherein the water content in the mixture C is 30-60%, the temperature is 25-45 ℃, and the fermentation time is 16-28h to form a mixture D; drying; and (5) mixing and crushing. The scheme can extract the protein peptide from the low-value trash fish and aquatic product leftovers, thereby reducing the processing cost of the protein peptide.

Description

Preparation method of fish protein peptide

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a preparation method of a fish protein peptide.

Background

Due to the rapid development of aquaculture and special aquaculture in China, the demand of animal protein feed is increased very rapidly, and the animal protein feed has the advantages of high protein content, high meat yield of animals, easiness in matching with other components and the like, but fish meal produced in China is generally low in grade, the processing technology is laggard, the fish meal price is low, and the problem that some fish meals with high protein content, such as special meal for prawns, special meal for turtles and the like, depend on import is serious.

At present, main economic fish resources in the ocean wither, and low-value small trash fishes cause insufficient fishing and processing due to various defects of the trash fishes. A large amount of leftovers (head, tail, bone and internal organs) are produced in the processing process of low-value small trash fish and aquatic products and are generally simply processed into fish powder or even discarded, so that resource waste and environmental pollution are caused. Therefore, it is necessary to research how to utilize the low-value small trash fish and aquatic product processing leftovers.

The small peptide is an intermediate product in the process of degrading protein into amino acid by animals, is an important nutrient substance of the animals, is one of important modes of nutrition absorption and metabolism of protein except the amino acid, has mutually independent absorption mechanisms with free amino acid, does not interfere with each other, is beneficial to the absorption of the amino acid and accelerates the synthesis of the protein. The peptide product is added into animal feed, so that the utilization rate of amino acid can be improved, the occurrence of diseases can be reduced, the production performance can be fully exerted, and the economic benefit can be improved.

At present, a plurality of preparation methods of bioactive peptides are available, an enzymolysis method and a microbial method are common, but the cost of external enzymes in the enzymolysis method is high, the enzymolysis time of small peptides prepared by the microbial method is too long, the cost is high, and after the conventional protease enzymolysis or microbial hydrolysis method is adopted, the maximum protein hydrolysis amount in raw materials can only reach 26%, wherein the content of polypeptide below 3000Da is only 83%, and the hydrolysis efficiency and the hydrolysis degree are difficult to realize industrial mass production.

Disclosure of Invention

The invention aims to provide a preparation method of fish protein peptide, which is used for extracting protein peptide from low-value small trash fish and aquatic product leftovers so as to reduce the processing cost of the protein peptide.

In order to achieve the above object, the basic scheme of the invention is as follows: a preparation method of fish protein peptide comprises the following steps:

raw material preparation: preparing raw materials, wherein the raw materials comprise marine low-value small trash fish and aquatic product leftovers;

a crushing step: putting the raw materials into a pulverizer, starting the pulverizer to pulverize, and pulverizing the raw materials into 5-10 meshes to form fish paste, and placing the fish paste in a raw material pool for later use;

an enzymolysis step: transferring the fish paste in the crushing step into an enzymolysis tank, and adding water into the enzymolysis tank, wherein the mass ratio of the water to the fish paste is 1: 1, mixing water and fish paste to form a mixture A, wherein the enzymolysis temperature is 50-70 ℃, the pH value during enzymolysis is 7.0, and the enzymolysis time is 2-6 h;

enzyme deactivation: heating the mixture A in the enzymolysis step to 90 ℃, and preserving the temperature for 10min to inactivate the enzyme to form a mixture B;

the preparation method of the small peptide liquid comprises the following steps: filtering the mixture B in the enzyme deactivation step, wherein the aperture of a filter screen is 40-100 meshes to form primary filter enzymatic hydrolysate and solid, and collecting the primary filter enzymatic hydrolysate and the solid respectively; then concentrating the primary filtered enzymatic hydrolysate to ensure that the water content of the primary filtered enzymatic hydrolysate is 30-70%, and collecting the enzymatic hydrolysate;

and (3) spray drying of the enzymolysis liquid: drying the enzymatic hydrolysate in the enzyme deactivation step by a spray dryer, controlling the pressure of the spray dryer to be 0.05-0.19Mpa, controlling the temperature at the inlet of the spray dryer to be 180-230 ℃ and the temperature at the outlet of the spray dryer to be 80-100 ℃, and obtaining dried enzymatic powder for later use;

and (3) microbial fermentation: mixing Aspergillus oryzae into the solid in the small peptide liquid preparation step to form a mixture C, wherein the water content in the mixture C is 30-60%, the temperature is 25-45 ℃, and the fermentation time is 16-28h to form a mixture D;

and (3) drying: taking out the mixture D in the microbial fermentation step, and drying the mixture D at a low temperature of 60-90 ℃ to form a dried material;

mixing and crushing: mixing the dried material with the enzymolysis powder in the enzymolysis liquid spray drying step to form a fish protein peptide crude product; carrying out micro-crushing treatment on the fish protein peptide to form a fish protein peptide finished product.

The principle and advantages of the basic scheme are as follows: 1. the raw materials prepared in the raw material preparation step comprise marine low-value trash fish and aquatic product leftovers, so that the two materials of the marine low-value trash fish and the aquatic product leftovers can be effectively utilized to prepare the fish protein peptide, and the yield of the whole protein peptide is increased; meanwhile, the marine low-value small trash fish and aquatic product leftovers are cheap and easy to obtain, and the processing cost of the protein peptide can be effectively reduced; meanwhile, the low-value marine fishes comprise anchovies, fringed shrimps, caper carps, carpesia and syngnathus, endogenous enzymes in the low-value marine fishes are developed, the stability is good, and the degradation of the macromolecular proteins of the fishes and the preparation of the protein peptides of the marine fishes are facilitated in the process of preparing the protein peptides; the fish types in the whole raw material are various, and the flavor and the nutritive value of the final fish protein peptide finished product can be effectively improved.

2. In the crushing step, the raw materials are ground to form fish paste, so that fish can be effectively reduced; the fish paste is convenient to use and carry out enzymolysis processing in an enzymolysis step, and is convenient to reduce rapid and efficient enzymolysis in an enzymolysis tank; meanwhile, because the marine low-value small trash fish and the aquatic product leftovers contain rich endogenous enzymes, the raw materials can be subjected to enzymolysis automatically, and the cost of adding exogenous enzymes is greatly reduced; the quality proportion of control water and raw materials can let abundant release of endogenous enzyme in this in-process, and the regulation of temperature and pH value can let endogenous enzyme's effect play the maximize simultaneously, realizes the quick enzymolysis of raw materials, adopts this in-process after, the hydrolysis of raw materials to the volume of protein can improve about 8%, and wherein below 3000Da polypeptide content can also increase about 8%.

3. The enzyme deactivation step is set for further fermentation in the microorganism fermentation step in the follow-up process, and meanwhile, the condition that the fermentation process of the endogenous enzyme on the fish paste cannot be controlled in the follow-up processing process can be avoided.

4. And a small peptide liquid preparation step, wherein the mixture A in the enzymolysis step can be filtered to form primary filter enzymolysis liquid and solid, the primary filter enzymolysis liquid can be further concentrated into enzymolysis liquid, the solid can be further fermented in the subsequent step, and the protein in the solid is refined into small peptides.

5. In the step of spray drying of the enzymatic hydrolysate, the enzymatic hydrolysate can be concentrated under high pressure, and the enzymatic hydrolysate is effectively concentrated into powder, so that the enzymatic hydrolysate can be conveniently mixed with a subsequent dried material and packaged; the inlet and the outlet of the spray dryer are controlled, so that the solid and the liquid in the enzymolysis liquid can be effectively separated, and the enzymolysis powder at the outlet can be conveniently collected.

6. In the microbial fermentation step, the aspergillus oryzae can further carry out microbial degradation on macromolecular protein in solid, so that the solid is decomposed into protein, the amount of the degradable solid is less in the process, the degradation speed of the whole raw material can be effectively improved, the bitter and fishy smell in the solid can be removed, and the follow-up eating is facilitated.

7. And a drying step, namely drying the mixture D after microbial fermentation, so as to be convenient for mixing with enzymolysis powder in the enzymolysis liquid spray drying step in the mixing and crushing step to form a fish protein peptide crude product, and then carrying out micro-crushing on the fish protein peptide crude product to form a fish protein peptide finished product, wherein the fish protein peptide finished product is convenient to eat.

In conclusion, the method adopts the marine low-value small trash fish and the aquatic product leftovers as the raw materials for preparing the fish protein peptide, and carries out enzymolysis on the raw materials by utilizing endogenous enzymes originally contained in the raw materials; after mixture B after preliminary enzymolysis progresses and filters, carries out microbial fermentation to the solid wherein again, carries out further degradation to the solid, and after two steps combine, can improve the content of the protein that the raw materials formation after hydrolysising, and then improve the fermentation efficiency of whole raw materials for the production efficiency of fish protein peptide, reduction in production cost.

Further, in the crushing step, before crushing the raw materials, the raw materials are firstly crushed into powder with the weight ratio of 1: 1-2, mixing with water, thawing, dispersing, and pulverizing.

Because the raw materials adopt the little trash fish of ocean low value and aquatic products leftover bits and pieces, it generally need freeze fresh-keeping when the transportation, mixes with water before smashing little trash fish of ocean low value and aquatic products leftover bits and pieces, is in order to unfreeze the raw materials, reduces subsequent crushing pressure, can improve the crushing efficiency of raw materials, and the raw materials of being convenient for form the fish paste fast.

Further, in the pulverization step, after the raw material is pulverized, the mesh number of the formed fish paste is not more than 10.

The mesh number of the fish paste is controlled to be within 10, so that the fish meat in the fish paste has small particles, rapid enzymolysis and microbial degradation can be conveniently carried out in subsequent steps, and the speed of decomposing raw materials into protein can be increased.

Further, in the enzymolysis step, powdery calcium chloride is added into the fish paste in the enzymolysis tank and uniformly mixed, so that the concentration of a calcium chloride solution in the fish paste is 0.02-0.03 mol/L.

In the enzymolysis step, calcium chloride is used for adjusting the pH value of the fish paste, so that the pH value of the fish paste during enzymolysis is 7.0, endogenous enzymes in the fish paste are in the optimal pH value, and the rapid enzymolysis of the endogenous enzymes on the fish paste is facilitated.

Further, in the enzymolysis step, when the pH value during enzymolysis is controlled, hydrochloric acid solution with the concentration of 1.0-2.0mol/L or sodium hydroxide solution with the concentration of 1.0-2.0mol/L is used for regulating the pH value.

In the enzymolysis process, the pH value of the fish paste also changes, and the pH value of the fish paste can be adjusted by adopting a hydrochloric acid solution or a sodium hydroxide solution, so that the activity of endogenous enzymes is kept all the time.

Further, in the enzymolysis step, the mixture A is stirred at the rotating speed of 40-70 r/min while enzymolysis is carried out.

When enzymolysis is carried out, the mixture A is stirred, so that endogenous enzymes in the mixture A can be conveniently and fully contacted with the mixture, and fish in fish paste can be conveniently and fully subjected to enzymolysis.

Further, in the step of spray drying the enzymatic hydrolysate, the pressure of a spray dryer is controlled to be 0.1-0.13Mpa, the temperature at the inlet of the spray dryer is 210 ℃, and the temperature at the outlet of the spray dryer is 90 ℃.

The temperature of the inlet and the outlet of the spray dryer is further controlled, and the temperature is adaptive to the temperature required by the enzymolysis liquid, so that the enzymolysis powder in the enzymolysis liquid is conveniently and quickly separated from the moisture, and the enzymolysis powder can be quickly dried.

Further, in the step of microbial fermentation, the water content in the mixture C is 40-50%, the temperature is 30-40 ℃, and the fermentation time is 18-24 h.

When the solid is subjected to microbial degradation, the microorganisms are placed in the mixture C with the water content of 40-50%, and the temperature is controlled within the range of 30-40 ℃, so that the microorganisms can rapidly metabolize, and the proteins in the mixture C are rapidly decomposed into small peptides.

Further, in the drying step, the temperature of the dried mixture D is low at 70-80 ℃.

The drying temperature is kept low, so that the small peptides or proteins in the mixture D can be prevented from deteriorating.

Further, in the enzymolysis step, the enzymolysis temperature is 55-60 ℃.

The control of the enzymolysis temperature is to provide a proper temperature for the endogenous enzyme, so that the endogenous enzyme can fully play a role.

Drawings

FIG. 1 is a flow chart of an embodiment of the method of the present invention for preparing a fish protein peptide according to example 1;

FIG. 2 is a graph showing the degree of hydrolysis of the raw materials in examples 1 to 4 when different hydrolysis periods were used.

Detailed Description

The following is further detailed by the specific embodiments:

example 1

The preparation method of the fish protein peptide in the embodiment 1 is basically as shown in the attached figure 1, and comprises the following specific steps:

raw material preparation: preparing raw materials including marine low-value small trash fish and aquatic product leftovers, wherein the marine low-value small trash fish mainly comprises anchovy, shrimp, capelin, carpesium and syngnathus; the aquatic product leftovers comprise fish heads, fish skins, fish tails and fish internal organs;

a crushing step: mixing the frozen low-value small trash fish and aquatic product leftovers with water, wherein the mass ratio of the raw materials to the water is 1: 2, unfreezing the raw materials by water, and fishing out after the raw materials are unfrozen and dispersed; putting the unfrozen raw materials into a pulverizer, starting the pulverizer to pulverize to form fish paste, enabling the mesh number of the fish paste to be less than or equal to 10, and then placing the fish paste in a raw material pool for standby;

an enzymolysis step: transferring the fish paste in the crushing step into an enzymolysis tank, and adding water into the enzymolysis tank, wherein the mass ratio of the water to the fish paste is 1: 1, mixing water and fish paste to form a mixture A; then adding powdery calcium chloride into the fish paste in the enzymolysis tank and uniformly mixing to ensure that the concentration of a calcium chloride solution in the fish paste is 0.025mol/L, then stirring the mixture A at a rotating speed of 50-60 r/min, fully performing enzymolysis on the fish paste, wherein in the enzymolysis process, the enzymolysis temperature is 55-60 ℃, the pH value during enzymolysis is 7.0, and the enzymolysis time is 2-6 h; in the enzymolysis process, when the pH value is more than 7, hydrochloric acid solution with the concentration of 1.0-2.0mol/L is used for adjusting the pH value of the mixture A; when the pH value is less than 7, adjusting the pH value of the mixture A by using a sodium hydroxide solution with the concentration of 1.0-2.0 mol/L;

the preparation method of the small peptide liquid comprises the following steps: filtering the mixture B in the enzyme deactivation step, wherein the aperture of a filter screen is 60-80 meshes to form primary filter enzymatic hydrolysate and solid, and collecting the primary filter enzymatic hydrolysate and the solid respectively; then concentrating the primary filtered enzymatic hydrolysate to ensure that the water content of the primary filtered enzymatic hydrolysate is 40-60%, and collecting the enzymatic hydrolysate;

and (3) spray drying of the enzymolysis liquid: drying the enzymatic hydrolysate in the enzyme deactivation step by a spray dryer, controlling the pressure of the spray dryer to be 0.1-0.13Mpa, controlling the temperature at the inlet of the spray dryer to be 210 ℃ and the temperature at the outlet of the spray dryer to be 90 ℃, and obtaining dried enzymatic hydrolysate powder for later use;

and (3) microbial fermentation: mixing Aspergillus oryzae into the solid in the small peptide liquid preparation step to form a mixture C, wherein the water content in the mixture C is 40-50%, the temperature is 30-40 ℃, and the fermentation time is 18h to form a mixture D; when the fermentation time is long, as shown in figure 2, the mass of the fish protein peptide obtained by hydrolyzing the raw material is 30% of the total mass of the raw material, so that the hydrolysis degree of the raw material in the prior art is effectively improved;

and (3) drying: taking out the mixture D in the microbial fermentation step, and drying the mixture D at a low temperature of 70-80 ℃ to form a dried material;

Example 2

Example 2 is different from example 1 in that, as shown in fig. 2, in the microbial fermentation step, the fermentation time is 36 hours, and after the microbial fermentation hydrolysis, the mass of the fish protein peptide obtained by hydrolysis from the raw material is 31% of the total mass of the raw material, which is improved compared with the hydrolysis degree of the raw material in example 1.

Example 3

Example 3 is different from example 1 in that, as shown in fig. 2, in the microbial fermentation step, the fermentation time period is 30 hours, and after the microbial fermentation hydrolysis, the mass of the fish protein peptide obtained by hydrolysis from the raw material is 32% of the total mass of the raw material, which is improved compared with the hydrolysis degree of the raw material in example 1.

Example 4

Example 4 is different from example 1 in that, as shown in fig. 2, in the microbial fermentation step, the fermentation time is 24 hours, after the microbial fermentation hydrolysis, the mass of the fish protein peptide obtained by hydrolysis from the raw material is 33% of the total mass of the raw material, which is the optimum time for the raw material to be hydrolyzed into protein, and the hydrolysis degree is the maximum value, so that the hydrolysis efficiency and the hydrolysis degree of the raw material can be effectively improved.

The foregoing is merely an example of the present invention and common general knowledge in the art of specific structures and/or features of the invention has not been set forth herein in any way. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.

Claims

1. A preparation method of a fish protein peptide is characterized by comprising the following steps:

an enzymolysis step: transferring the fish paste in the crushing step into an enzymolysis tank, and adding water into the enzymolysis tank, wherein the mass ratio of the water to the fish paste is 1: 1, mixing water and fish paste to form a mixture A, wherein the enzymolysis temperature is 50-70 ℃, the pH during enzymolysis is =7.0, and the enzymolysis time is 2-6 h;

enzyme deactivation: heating the mixture A in the enzymolysis step to 90 ℃, and preserving heat for 5-15min to inactivate enzyme to form a mixture B;

2. The method according to claim 1, wherein the raw materials are pulverized in a weight ratio of 1: 1-2, mixing with water, thawing, dispersing, and pulverizing.

3. The method according to claim 2, wherein the fish protein peptide is obtained by pulverizing the raw material to obtain a fish paste having a mesh size of 10 or less.

4. The method for preparing fish protein peptide according to claim 3, wherein in the enzymolysis step, powdery calcium chloride is added into the fish paste in the enzymolysis tank and uniformly mixed, so that the concentration of the calcium chloride solution in the fish paste is 0.02-0.03 mol/L.

5. The method according to claim 4, wherein the pH value is adjusted by a hydrochloric acid solution with a concentration of 1.0-2.0mol/L or a sodium hydroxide solution with a concentration of 1.0-2.0mol/L during the enzymolysis step.

6. The method for preparing fish protein peptide according to claim 5, wherein in the enzymolysis step, the mixture A is stirred at a rotating speed of 40-70 r/min while enzymolysis is carried out.

7. The method for preparing a fish protein peptide according to claim 5, wherein in the step of spray-drying the enzymatic hydrolysate, the pressure of the spray-dryer is controlled to be 0.1-0.13MPa, the temperature at the inlet of the spray-dryer is 210 ℃, and the temperature at the outlet of the spray-dryer is 90 ℃.

8. The method for preparing fish protein peptide according to claim 6, wherein the water content in the mixture C in the step of microbial fermentation is 40-50%, the temperature is 30-40 ℃, and the fermentation time is 18-24 h.

9. The method of claim 7, wherein the temperature of the oven-dried mixture D is 70-80 ℃ in the oven-drying step.

10. The method according to claim 8, wherein the enzymolysis temperature is 55-60 ℃.