Freezing denaturation resisting agent for golden tiger hybrid spotted fish meat protein and preparation method of freezing denaturation resisting agent

Abstract

The invention provides an anti-freezing denaturation agent for golden and tiger hybrid spotted fish meat protein and a preparation method of the anti-freezing denaturation agent. The invention relates to an anti-freezing denaturation agent for protein of Jinhu hybrid spotted fish. The anti-freezing denaturation agent is prepared from the following raw materials in parts by weight: 5 to 8 parts of trehalose-mannitol, 0.5 to 1 part of nano chitosan, 0.2 to 0.5 part of rosmarinic acid and 0.1 to 0.2 part of polysorbate-80. The trehalose-mannitol is prepared from trehalose and mannitol. Aiming at the characteristics of Jinhu hybrid spotted fish meat, trehalose-mannitol, nano chitosan, rosmarinic acid and polysorbate-80 are taken as raw materials and compounded to obtain a new formula of the protein anti-freezing denaturation agent for the Jinhu hybrid spotted fish meat, and the anti-freezing denaturation agent prepared by adopting the formula and the process disclosed by the invention can be used for inhibiting protein degradation and improving the anti-freezing denaturation effect of the Jinhu hybrid spotted fish meat. The decrease range of MP protein concentration, MP total sulfydryl content and MP Ca < 2 + >-ATPase activity of the antifreeze agent is obviously reduced, and the effect of the antifreeze agent is superior to that of a commercially available antifreeze agent. Moreover, the anti-freezing denaturation agent for the protein of the Jinhu hybrid spotted fish does not contain phosphorus, and has the advantages of safety and high efficiency.

Description

Technical Field

[0001] The invention relates to the field of grouper, and in particular to a golden tiger hybrid grouper meat protein anti-freezing denaturant and a preparation method thereof. Background Art

[0002] Golden Tiger hybrid grouper is a high-quality new species of grouper bred in recent years, characterized by rapid growth, strong disease resistance, and delicious meat. Myofibrillar protein accounts for up to 70% of its muscle. During freezing, ice crystal formation and oxidation easily disrupt the protein's spatial structure, leading to a decrease in water retention, a hardened texture, and a loss of flavor, severely restricting its export and advanced processing.

[0003] Existing technologies are mostly designed for common grouper or freshwater fish. However, due to hybrid advantages, the golden tiger hybrid grouper has a unique protein composition, and general antifreeze is difficult to adapt to its characteristics. Existing formulas lack targeting.

[0004] With the development of cold chain logistics, the market urgently needs protein anti-freeze denaturants that can maximize the nutritional value and taste of hybrid grouper. However, there is currently a lack of specialized anti-freeze denaturants tailored to the specific characteristics of hybrid grouper protein, and existing solutions have significant shortcomings in adaptability. The present invention is of great significance for the development of an efficient and safe anti-freeze denaturant specifically for hybrid grouper meat. Summary of the Invention

[0005] In view of this, the present invention provides a golden tiger hybrid grouper meat protein anti-freezing denaturant and a preparation method thereof.

[0006] The technical solution of the present invention is achieved as follows:

[0007] A golden tiger hybrid grouper meat protein anti-freezing denaturation agent comprises the following raw materials in parts by weight: 5-8 parts of trehalose-mannitol, 0.5-1 part of nano-chitosan, 0.2-0.5 part of rosmarinic acid, and 0.1-0.2 part of polysorbate 80; the trehalose-mannitol is prepared from trehalose and mannitol.

[0008] Preferably, the anti-freezing denaturation agent for hybrid grouper fish meat protein comprises the following raw materials in parts by weight: 7 parts of trehalose-mannitol, 0.8 parts of nano-chitosan, 0.3 parts of rosmarinic acid, and 0.15 parts of polysorbate 80.

[0009] Preferably, the trehalose-mannitol is prepared from trehalose and mannitol in a mass ratio of 3-5:1.

[0010] Preferably, the preparation method of trehalose-mannitol comprises the following steps: mixing trehalose and mannitol with 45-55°C deionized water at a material-liquid ratio of 1:2.5-3.5 (kg / L), cooling the mixture to 28-32°C at a rate of 0.4-0.6°C / min, and then cooling the mixture to 3-5°C at a rate of 0.9-1.1°C / min to obtain trehalose-mannitol.

[0011] Preferably, the particle size of the nano-chitosan is 100-150 nm.

[0012] The method for preparing the anti-freezing denaturation agent of golden tiger hybrid grouper meat protein of the present invention comprises the following steps:

[0013] (1) Nano-chitosan was mixed with 0.8%-1.2% v / v acetic acid solution to prepare a chitosan solution with a mass concentration of 1%-2%; the chitosan solution was sprayed onto trehalose-mannitol by a fluidized bed coating process to obtain a complex A;

[0014] (2) Rosmarinic acid and polysorbate 80 were mixed, 4%-6% v / v ethanol solution was added at a material-liquid ratio of 1:1.5-2.0, high pressure homogenization was performed, and the solvent was removed by rotary evaporation to obtain complex B;

[0015] (3) Mixing the complex A and the complex B, stirring and mixing, to obtain the target anti-freeze denaturing agent.

[0016] Preferably, in step (1), the fluidized bed coating process conditions are: inlet air temperature 58-62°C, air volume 24-26Hz, atomization pressure 0.5-0.6MPa, and spray rate 25-30ml / min.

[0017] Preferably, in step (2), the pressure of the high-pressure homogenization is 150-200 MPa, and the homogenization time is 3-8 min.

[0018] Preferably, in step (3), the stirring speed is 20-30 rpm and the stirring time is 20-40 min.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention aims at the characteristics of golden tiger hybrid grouper meat, uses trehalose-mannitol, nano-chitosan, rosmarinic acid, and polysorbate-80 as raw materials, and compounded to obtain a new formula of anti-freezing denaturant for golden tiger hybrid grouper meat protein. The anti-freezing denaturant prepared by the formula and process of the present invention can inhibit protein degradation, and its MP protein concentration, MP total thiol content, MP Ca 2+The decline in ATPase activity is significantly reduced, and the effect is superior to commercially available antifreeze agents, better preserving the flavor of the golden tiger hybrid grouper meat. Furthermore, the present invention's golden tiger hybrid grouper meat protein antifreeze denaturant exhibits excellent water retention, which helps preserve the flavor of the golden tiger hybrid grouper meat. Furthermore, the present invention's golden tiger hybrid grouper meat protein antifreeze denaturant is phosphorus-free, offering the advantages of safety and effectiveness. DETAILED DESCRIPTION

[0021] In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention.

[0022] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.

[0023] Unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources.

[0024] Example 1

[0025] A golden tiger hybrid grouper meat protein anti-freezing denaturation agent comprises the following raw materials in parts by weight: 7 parts of trehalose-mannitol, 0.8 parts of nano-chitosan, 0.3 parts of rosmarinic acid, and 0.15 parts of polysorbate 80; the particle size of the nano-chitosan is 100-150 nm; the trehalose-mannitol is prepared from trehalose and mannitol in a mass ratio of 4:1; and the preparation method of the trehalose-mannitol comprises the following steps: mixing trehalose and mannitol with 50°C deionized water at a material-liquid ratio of 1:3.0 (kg / L), cooling the mixture to 30°C at a rate of 0.5°C / min, and then cooling the mixture to 4°C at a rate of 1.0°C / min, to obtain the trehalose-mannitol.

[0026] The method for preparing the above-mentioned anti-freezing denaturation agent for golden tiger hybrid grouper meat protein comprises the following steps:

[0027] (1) Nano-chitosan was mixed with 1.0% v / v acetic acid solution to prepare a chitosan solution with a mass concentration of 1.5%; the chitosan solution was sprayed onto trehalose-mannitol using a fluidized bed coating process (inlet air temperature 60°C, air volume 25 Hz, atomization pressure 0.55 MPa, spray rate 28 ml / min) to obtain complex A;

[0028] (2) Rosmarinic acid and polysorbate 80 were mixed, and 5% v / v ethanol solution was added at a material-liquid ratio of 1:1.8. The mixture was homogenized under high pressure (homogenization pressure of 180 MPa, homogenization time of 5 min), and the solvent was removed by rotary evaporation to obtain complex B;

[0029] (3) Compound A and compound B were mixed and stirred (stirring speed was 25 rpm, stirring time was 30 min) to obtain the target anti-freeze denaturant.

[0030] Example 2

[0031] A golden tiger hybrid grouper meat protein anti-freezing denaturation agent comprises the following raw materials in parts by weight: 5 parts of trehalose-mannitol, 0.5 parts of nano-chitosan, 0.2 parts of rosmarinic acid, and 0.1 parts of polysorbate 80; the particle size of the nano-chitosan is 100-150 nm; the trehalose-mannitol is prepared from trehalose and mannitol in a mass ratio of 3:1; and the preparation method of the trehalose-mannitol comprises the following steps: mixing trehalose and mannitol with 50°C deionized water at a material-liquid ratio of 1:3.5 (kg / L), cooling the mixture to 30°C at a rate of 0.5°C / min, and then cooling the mixture to 4°C at a rate of 1.0°C / min, to obtain the trehalose-mannitol.

[0032] The preparation method of the above-mentioned golden tiger hybrid grouper meat protein anti-freezing denaturant is the same as that in Example 1.

[0033] Example 3

[0034] A golden tiger hybrid grouper meat protein anti-freezing denaturation agent comprises the following raw materials in parts by weight: 8 parts of trehalose-mannitol, 1.0 part of nano-chitosan, 0.5 part of rosmarinic acid, and 0.2 part of polysorbate 80; the particle size of the nano-chitosan is 100-150 nm; the trehalose-mannitol is prepared from trehalose and mannitol in a mass ratio of 5:1; and the preparation method of the trehalose-mannitol comprises the following steps: mixing trehalose and mannitol with 50°C deionized water at a material-liquid ratio of 1:2.5 (kg / L), cooling the mixture to 30°C at a rate of 0.5°C / min, and then cooling the mixture to 4°C at a rate of 1.0°C / min, to obtain the trehalose-mannitol.

[0035] The preparation method of the above-mentioned golden tiger hybrid grouper meat protein anti-freezing denaturant is the same as that in Example 1.

[0036] Comparative Example 1

[0037] The difference from Example 1 is that mannitol is replaced with sorbitol. A golden tiger hybrid grouper meat protein anti-freeze denaturation agent comprises the following raw materials in parts by weight: 7 parts trehalose-sorbitol, 0.8 parts nano-chitosan, 0.3 parts rosmarinic acid, and 0.15 parts polysorbate 80; the nano-chitosan has a particle size of 100-150 nm. The trehalose-sorbitol is prepared from trehalose and sorbitol in a mass ratio of 4:1. The preparation method of the trehalose-sorbitol comprises the following steps: mixing trehalose and sorbitol with 50°C deionized water at a material-liquid ratio of 1:3.0 kg / L, cooling the mixture to 30°C at a rate of 0.5°C / min, and then cooling the mixture to 4°C at a rate of 1.0°C / min, to obtain the trehalose-sorbitol.

[0038] Comparative Example 2

[0039] The difference from Example 1 is that nano-chitosan is replaced by carboxymethyl chitosan. A golden tiger hybrid grouper meat protein anti-freezing denaturant comprises the following raw materials in parts by weight: 7 parts of trehalose-mannitol, 0.8 parts of carboxymethyl chitosan, 0.3 parts of rosmarinic acid, and 0.15 parts of polysorbate 80.

[0040] Comparative Example 3

[0041] The difference from Example 1 is that rosmarinic acid is replaced by tea polyphenols. A golden tiger hybrid grouper meat protein anti-freezing denaturant comprises the following raw materials in parts by weight: 7 parts of trehalose-mannitol, 0.8 parts of nano-chitosan, 0.3 parts of tea polyphenols, and 0.15 parts of polysorbate 80.

[0042] Comparative Example 4

[0043] The difference from Example 1 is that the preparation method of trehalose-mannitol includes the following steps: mixing trehalose and mannitol with 50°C deionized water at a material-liquid ratio of 1:3.0 in kg / L, and directly cooling to 4°C (without controlling the cooling rate) to obtain trehalose-mannitol.

[0044] Comparative Example 5

[0045] The difference from Example 1 is that the nano-chitosan uses 80-90 nm nano-chitosan. A golden tiger hybrid grouper meat protein anti-freeze denaturation agent comprises the following raw materials in parts by weight: 7 parts trehalose-mannitol, 0.8 parts nano-chitosan, 0.3 parts rosmarinic acid, and 0.15 parts polysorbate 80; the nano-chitosan has a particle size of 80-90 nm.

[0046] Test Example 1

[0047] Study Groups 1-8: Anti-freeze denaturation agents of Examples 1-3 and Comparative Examples 1-5.

[0048] Positive control group: food-grade compound phosphate (commercially available antifreeze, Henan Zhongchen Biotechnology Co., Ltd.);

[0049] Blank group: no treatment.

[0050] Fresh golden tiger hybrid grouper meat was prepared into surimi to evaluate its antifreeze activity.

[0051] The dorsal muscles of the golden tiger hybrid spot were minced into minced fish using a meat grinder, rinsed three times with deionized water, centrifuged at 10,000 r / min for 10 min, and the precipitate was collected to obtain minced fish.

[0052] In the study groups 1-8, 0.5% by mass of the antifreeze denaturant of Examples 1-3 and Comparative Examples 1-5 was added to the surimi; the positive control group used 0.5% by mass of a commercial antifreeze (complex phosphate) added to the surimi, and the blank group was not treated; after mixing evenly, the mixture was placed at -20°C for freezing and thawing at room temperature with running water (deionized water) for 1 hour, and the freeze-thaw cycle was repeated 5 times, with each cycle lasting 3 days. After 5 freeze-thaw cycles, the surimi myofibrillar protein (MP) was extracted according to the conventional method, and the surimi MP protein concentration, surimi MP total thiol content, surimi MP Ca content were determined. 2+ -ATPase activity was compared with that on day 0, and the percentage decrease of each index after 5 freeze-thaw cycles was calculated. The results are shown in Table 1 below.

[0053] Among them, the protein concentration of surimi MP was determined by using a biuret protein quantitative kit, and the total thiol content of surimi MP was determined by using a total thiol content kit. 2+ -ATPase activity using ultra-trace Ca 2 + -ATPase assay kit.

[0054] Table 1 MP protein concentration, MP total thiol content, MP Ca 2+ -Decrease in ATPase activity

[0055]

[0056] The above results show that, compared with the blank group, the use of the anti-freeze denaturant of Examples 1-3 is beneficial to inhibiting protein degradation, and the MP protein concentration, MP total thiol content, MP Ca 2+ -ATPase activity decrease percentage is significantly reduced, and the effect is better than commercial antifreeze. Compared with Example 1, the decrease rate of Comparative Examples 1-4 is significantly increased; and compared with Example 1, the decrease rate of Comparative Example 5 is also increased.

[0057] Example 4

[0058] The difference from Example 1 is: the preparation method of the anti-freezing denaturant of golden tiger hybrid grouper meat protein.

[0059] (1) Nano-chitosan was mixed with 1.0% v / v acetic acid solution to prepare a chitosan solution with a mass concentration of 1%; the chitosan solution was sprayed onto trehalose-mannitol using a fluidized bed coating process (inlet air temperature 60°C, air volume 25 Hz, atomization pressure 0.55 MPa, spray rate 28 ml / min) to obtain complex A;

[0060] (2) Rosmarinic acid and polysorbate 80 were mixed, and 5% v / v ethanol solution was added at a material-liquid ratio of 1:1.5. The mixture was homogenized under high pressure (homogenization pressure of 150 MPa, homogenization time of 8 min), and the solvent was removed by rotary evaporation to obtain complex B;

[0061] (3) Compound A and compound B were mixed and stirred (stirring speed was 20 rpm, stirring time was 40 min) to obtain the target anti-freeze denaturant.

[0062] Example 5

[0063] The difference from Example 1 is: the preparation method of the anti-freezing denaturant of golden tiger hybrid grouper meat protein.

[0064] (1) Nano-chitosan was mixed with 1.0% v / v acetic acid solution to prepare a chitosan solution with a mass concentration of 2%; the chitosan solution was sprayed onto trehalose-mannitol using a fluidized bed coating process (inlet air temperature 60°C, air volume 25 Hz, atomization pressure 0.55 MPa, spray rate 28 ml / min) to obtain complex A;

[0065] (2) Rosmarinic acid and polysorbate 80 were mixed, and 5% v / v ethanol solution was added at a material-liquid ratio of 1:2.0. The mixture was homogenized under high pressure (homogenization pressure of 200 MPa, homogenization time of 3 min), and the solvent was removed by rotary evaporation to obtain complex B;

[0066] (3) Compound A and compound B were mixed and stirred (stirring speed was 30 rpm, stirring time was 20 min) to obtain the target anti-freeze denaturant.

[0067] The anti-freezing denaturant prepared in Example 4-5 was used for freezing and preserving the meat of hybrid grouper fish, and had a good anti-freezing denaturation effect.

[0068] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A golden tiger hybrid grouper meat protein anti-freezing denaturant, characterized in that: The invention comprises the following raw materials in parts by weight: 5-8 parts of trehalose-mannitol, 0.5-1 part of nano-chitosan, 0.2-0.5 part of rosmarinic acid and 0.1-0.2 part of polysorbate 80; the trehalose-mannitol is prepared from trehalose and mannitol.

2. The anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 1, characterized in that The trehalose-mannitol is prepared from trehalose and mannitol in a mass ratio of 3-5:

1.

3. The anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 2, characterized in that The preparation method of trehalose-mannitol comprises the following steps: mixing trehalose and mannitol with 45-55°C deionized water at a material-liquid ratio of 1:2.5-3.5 (kg / L), cooling the mixture to 28-32°C at a rate of 0.4-0.6°C / min, and then cooling the mixture to 3-5°C at a rate of 0.9-1.1°C / min to obtain trehalose-mannitol.

4. The anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 1, characterized in that The particle size of the nano-chitosan is 100-150 nm.

5. The anti-freezing denaturant of golden tiger hybrid grouper meat protein according to any one of claims 1 to 4, characterized in that: The invention comprises the following raw materials in parts by weight: 7 parts of trehalose-mannitol, 0.8 parts of nano-chitosan, 0.3 parts of rosmarinic acid and 0.15 parts of polysorbate 80.

6. The method for preparing the anti-freezing denaturant of golden tiger hybrid grouper meat protein according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) Nano-chitosan was mixed with 0.8%-1.2% v / v acetic acid solution to prepare a chitosan solution with a mass concentration of 1%-2%; the chitosan solution was sprayed onto trehalose-mannitol by a fluidized bed coating process to obtain a complex A; (2) Rosmarinic acid and polysorbate 80 were mixed, 4%-6% v / v ethanol solution was added at a material-liquid ratio of 1:1.5-2.0, high pressure homogenization was performed, and the solvent was removed by rotary evaporation to obtain complex B; (3) Mixing the complex A and the complex B, stirring and mixing, to obtain the target anti-freeze denaturing agent.

7. The method for preparing the anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 6, characterized in that: Step (1), the fluidized bed coating process conditions are: inlet air temperature 58-62°C, air volume 24-26Hz, atomization pressure 0.5-0.6MPa, and spray rate 25-30ml / min.

8. The method for preparing the anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 6 or 7, characterized in that: In step (2), the pressure of the high-pressure homogenization is 150-200 MPa, and the homogenization time is 3-8 min.

9. The method for preparing the anti-freezing denaturant of golden tiger hybrid grouper meat protein according to claim 6, characterized in that: In step (3), the stirring speed is 20-30 rpm and the stirring time is 20-40 min.

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