Animal enzyme and its preparation method and application

Abstract

The present invention relates to the field of medical technology, and in particular to an animal enzyme, a preparation method thereof and an application thereof. The animal enzyme comprises, by weight: animal source protein I: 40-60 parts; animal source protein II comprises: black soldier fly larvae: 10-20 parts, silkworm pupae: 10-20 parts; flaxseed: 4-8 parts; honeysuckle: 4-8 parts; Viola yedoensis: 4-8 parts; compound enzyme comprises: acid protease: 0.2-0.6 parts, lipase: 0.1-0.3 parts, cellulase: 0.1-0.3 parts; fermentation agent I comprises: Bacillus amyloliquefaciens 0.5-2 parts, Bacillus subtilis 0.5 0.2-1 parts; fermentation agent II comprises: Lactobacillus plantarum 1 0.5-2 parts, Lactobacillus acidophilus 0.5 0.2-1 parts; sugar: 1-3 parts; water: 80-120 parts; the animal enzyme in the present invention has a certain direct inactivation effect on feline calicivirus FCV virus.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an animal enzyme and a preparation method and application thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Viral infection is one of the major risk factors that endanger the health and even life of pets. Felinecalicivirus (FCV) is a pathogen that causes respiratory diseases in cats. It belongs to the Caliciviridae family and the Vesivirus genus. The disease is widely spread among cats. It has a high risk of disease, is difficult to cure, and has a high mortality rate, especially for kittens, where the mortality rate can reach 67%. At present, the treatment for this disease mainly uses interferon, broad-spectrum antiviral drugs, antibiotics, anti-inflammatory drugs, etc., but the treatment effect is not very ideal.

[0004] Animal enzymes are products containing specific biologically active ingredients that are made from animal protein as the main raw material and fermented by microorganisms. At present, most animal enzymes used for feeding use only use animal protein as the main raw material, and are mostly used for feeding poultry (such as chickens) and livestock (cattle, pigs, etc.) to improve the immunity, feed-to-meat ratio, live piglet rate, healthy piglet rate, etc. of animals, such as CN 111838408 A. However, there are no reports on animal enzymes for pets, especially cats and their viral infections. Summary of the invention

[0005] Based on the problems existing in the prior art, the present invention uses a certain proportion of black soldier fly larvae and silkworm pupae as animal protein sources, combined with flaxseed, honeysuckle and Viola yedoensis Chinese medicinal ingredients, and selects a variety of probiotics for mixed fermentation to prepare animal enzymes with certain antiviral activity. In view of this, the purpose of the present invention is to provide an animal enzyme and its preparation method and application.

[0006] Specifically, the technical solution of the present invention is as follows:

[0007] In a first aspect of the present invention, an animal enzyme is provided, which is made from the following raw materials in parts by weight:

[0008] Animal source protein I: 40~60 parts;

[0009] Animal source protein II includes: black soldier fly larvae: 10-20 parts, silkworm pupae: 10-20 parts;

[0010] Flaxseed: 4-8 parts; Honeysuckle: 4-8 parts; Viola yedoensis: 4-8 parts;

[0011] The compound enzyme includes: acid protease: 0.2-0.6 parts, lipase: 0.1-0.3 parts, cellulase: 0.1-0.3 parts;

[0012] The fermentation agent I comprises: 0.5-2 parts of Bacillus amyloliquefaciens and 0.5-0.2-1 parts of Bacillus subtilis;

[0013] The fermentation agent II includes: 0.5-2 parts of Lactobacillus plantarum 1 and 0.5-0.2-1 parts of Lactobacillus acidophilus;

[0014] Sugar: 1~3 parts;

[0015] Water: 80~120 parts;

[0016] The animal source protein I is one or more of livestock meat or poultry meat.

[0017] In a specific embodiment, the animal enzyme is made from the following raw materials in parts by weight:

[0018] Animal source protein I: 45-55 parts;

[0019] Animal source protein II includes: black soldier fly larvae: 12-18 parts, silkworm pupae: 12-18 parts;

[0020] Flaxseed: 5-7 parts; Honeysuckle: 5-7 parts; Viola yedoensis: 5-7 parts;

[0021] Compound enzyme: acid protease: 0.2~0.4 parts, lipase: 0.1~0.3 parts, cellulase: 0.1~0.3 parts;

[0022] The fermentation agent I comprises: 0.8-1.2 parts of Bacillus amyloliquefaciens and 0.3-0.8 parts of Bacillus subtilis;

[0023] The fermentation agent II includes: 0.8-1.2 parts of Lactobacillus plantarum and 0.3-0.8 parts of Lactobacillus acidophilus;

[0024] Sugar: 1~2 parts;

[0025] Water: 90~110 parts.

[0026] In a specific embodiment, the sugar is selected from glucose, starch or a combination thereof.

[0027] In a second aspect of the present invention, a method for preparing the animal enzyme according to the first aspect is provided, comprising the following steps:

[0028] (1) Mix the ground flaxseed, honeysuckle, Viola yedoensis, and the ground animal protein I and animal protein II with water, and then sterilize them;

[0029] (2) Add sugar to the sterilized material in step (1) and mix well;

[0030] (3) Adding the complex enzyme to the material treated in step (2), mixing well, and performing enzymolysis at 30-37°C for 0.5-2 h;

[0031] (4) adjusting the pH of the material treated in step (3) to 7.0-8.0, inoculating fermentation agent I, and aerobically fermenting at 30-37°C for 20-30 h;

[0032] (5) Add fermentation agent II to the material after fermentation in step (4), and perform anaerobic fermentation at 30-37°C for 24-48 hours.

[0033] In a specific embodiment, in step (1), the mesh size of the flaxseed, honeysuckle and Viola yedoensis after pulverization is 50-100 mesh; the mesh size of the animal source protein I and animal source protein II after refining is 100-200 mesh.

[0034] In a specific embodiment, in step (1), the sterilization temperature is 80-100° C. and the time is 1-4 hours.

[0035] In a specific embodiment, in step (3), the enzymatic hydrolysis temperature is preferably 35-37°C.

[0036] In a specific embodiment, in step (4), the pH of the material treated in step (3) is adjusted to 7.4-7.5.

[0037] In a specific embodiment, in step (4), the temperature of aerobic fermentation is preferably 35-37° C., and the fermentation time is preferably 20-28 h.

[0038] In a specific embodiment, in step (5), the temperature of anaerobic fermentation is preferably 35-37° C., and the fermentation time is preferably 24-36 h.

[0039] In the third aspect of the present invention, there is provided a use of the animal enzyme described in the first aspect in the preparation of animal antiviral drugs.

[0040] Preferably, the animal is a cat.

[0041] Preferably, the virus is feline calicivirus (FCV).

[0042] The present invention has the following beneficial effects:

[0043] The animal enzyme in the present invention contains a certain proportion of black soldier fly larvae and silkworm pupae as animal protein sources, combined with flaxseed, honeysuckle and Viola yedoensis Chinese medicinal ingredients, and a variety of probiotics are selected for mixed fermentation. The obtained animal enzyme has a certain direct inactivation effect on feline calicivirus (FCV). DETAILED DESCRIPTION

[0044] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used in the present application have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0045] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0046] The following examples are only used to illustrate the embodiments of the present invention and do not constitute any limitation to the present invention. Those skilled in the art will appreciate that various modifications may be made without departing from the spirit and scope of the present invention, and these modifications all fall within the scope of protection of the present invention.

[0047] Bacteria and sources:

[0048] Bacillus amyloliquefaciens, Bacillus subtilis, Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus licheniformis were purchased from ATCC.

[0049] Example 1 Preparation of Animal Enzyme 1

[0050] The raw materials of the animal enzyme in this embodiment include the following in parts by weight:

[0051] Animal source protein I: 50 parts;

[0052] Animal source protein II includes: black soldier fly larvae: 15 parts, silkworm pupae: 15 parts;

[0053] Flaxseed: 6 parts; Honeysuckle: 6 parts; Viola yedoensis: 6 parts;

[0054] The compound enzyme includes: acid protease: 0.3 parts, lipase: 0.2 parts, cellulase: 0.2 parts;

[0055] The fermentation agent I comprises: 1 part of Bacillus amyloliquefaciens and 0.5 part of Bacillus subtilis;

[0056] The fermentation agent II includes: 1 part of Lactobacillus plantarum and 0.5 part of Lactobacillus acidophilus;

[0057] Sugar: 1.5 parts;

[0058] Water: 100 parts.

[0059] Preparation method of animal enzyme 1:

[0060] (1) Animal source protein I (pork, chicken, duck in a ratio of 1:1:1) and animal source protein II were crushed and ground to about 140 mesh; flaxseed, honeysuckle, and Viola yedoensis were crushed to 80 mesh, mixed with water, and sterilized at 80°C for 2 h;

[0061] (2) Add sugar to the sterilized material in step (1) and stir evenly;

[0062] (3) Add the complex enzyme to the material treated in step (2), stir evenly, and perform enzymolysis at 37°C for 1 h;

[0063] (4) adjusting the pH value of the material after the treatment in step (3) to 7.4-7.5, inoculating the fermentation agent I, and performing aerobic fermentation at 37°C under stirring conditions, and taking out part of the material at 16 h, 20 h, 24 h, 28 h, 32 h, and 36 h for the next step;

[0064] (5) adding fermentation agent II to the material after fermentation in step (4), and performing anaerobic fermentation at 30-37°C for 36 h;

[0065] (6) The material after anaerobic fermentation in step (5) is crushed to 200 mesh to obtain animal enzymes 1a-1f.

[0066] Example 2 Preparation of Animal Enzyme 2

[0067] The raw materials of the animal enzyme in this embodiment include the following in parts by weight:

[0068] Animal source protein I: 60 parts;

[0069] Animal source protein II includes: black soldier fly larvae: 10 parts, silkworm pupae: 10 parts;

[0070] Flaxseed: 4 parts; Honeysuckle: 6 parts; Viola yedoensis: 8 parts;

[0071] The compound enzyme includes: acid protease: 0.3 parts, lipase: 0.2 parts, cellulase: 0.2 parts;

[0072] The fermentation agent I comprises: 1 part of Bacillus amyloliquefaciens and 0.5 part of Bacillus subtilis;

[0073] The fermentation agent II includes: 1 part of Lactobacillus plantarum and 0.5 part of Lactobacillus acidophilus;

[0074] Sugar: 1.5 parts;

[0075] Water: 100 parts.

[0076] Preparation method of animal enzyme 2:

[0077] (1) Animal source protein I (pork, chicken, duck in a ratio of 1:1:1) and animal source protein II were crushed and ground to about 140 mesh; flaxseed, honeysuckle, and Viola yedoensis were crushed to 80 mesh, mixed with water, and sterilized at 80°C for 2 h;

[0078] (2) Add sugar to the sterilized material in step (1) and stir evenly;

[0079] (3) Add the complex enzyme to the material treated in step (2), stir evenly, and perform enzymolysis at 37°C for 1 h;

[0080] (4) adjusting the pH value of the material after treatment in step (3) to 7.4-7.5, inoculating fermentation agent I, and performing aerobic fermentation at 37°C with stirring for 24 hours;

[0081] (5) adding fermentation agent II to the material after fermentation in step (4), and performing anaerobic fermentation at 30-37°C for 36 hours;

[0082] (6) The material after anaerobic fermentation in step (5) is crushed to 200 mesh to obtain animal enzyme 2.

[0083] Example 3 Preparation of Animal Enzyme 3

[0084] The raw materials of the animal enzyme in this embodiment include the following in parts by weight:

[0085] Animal source protein I: 40 parts;

[0086] Animal source protein II includes: black soldier fly larvae: 20 parts, silkworm pupae: 20 parts;

[0087] Flaxseed: 8 parts; Honeysuckle: 6 parts; Viola yedoensis: 4 parts;

[0088] The compound enzyme includes: acid protease: 0.3 parts, lipase: 0.2 parts, cellulase: 0.2 parts;

[0089] The fermentation agent I comprises: 1 part of Bacillus amyloliquefaciens and 0.5 part of Bacillus subtilis;

[0090] The fermentation agent II includes: 1 part of Lactobacillus plantarum and 0.5 part of Lactobacillus acidophilus;

[0091] Sugar: 1.5 parts;

[0092] Water: 100 parts.

[0093] Preparation method of animal enzyme 3:

[0094] (1) Animal source protein I (pork, chicken, duck in a ratio of 1:1:1) and animal source protein II were crushed and ground to about 140 mesh; flaxseed, honeysuckle, and Viola yedoensis were crushed to 80 mesh, mixed with water, and sterilized at 80°C for 2 h;

[0095] (2) Add sugar to the sterilized material in step (1) and stir evenly;

[0096] (3) Add the complex enzyme to the material treated in step (2), stir evenly, and perform enzymolysis at 37°C for 1 h;

[0097] (4) adjusting the pH value of the material after treatment in step (3) to 7.4-7.5, inoculating fermentation agent I, and performing aerobic fermentation at 37°C with stirring for 24 hours;

[0098] (5) adding fermentation agent II to the material after fermentation in step (4), and performing anaerobic fermentation at 30-37°C for 36 h;

[0099] (6) The material after anaerobic fermentation in step (5) is crushed to 200 meshes to obtain animal enzyme 3.

[0100] Comparative Example 1: Only animal protein I is used as raw material

[0101] The raw materials of the animal enzyme in this comparative example are as follows, in parts by weight, including:

[0102] Animal source protein I: 98 parts;

[0103] The complex enzyme includes: acid protease: 0.3 parts, lipase: 0.2 parts;

[0104] The fermentation agent I comprises: 1 part of Bacillus amyloliquefaciens and 0.5 part of Bacillus subtilis;

[0105] The fermentation agent II includes: 1 part of Lactobacillus plantarum and 0.5 part of Lactobacillus acidophilus;

[0106] Sugar: 1.5 parts;

[0107] Water: 100 parts.

[0108] Preparation method of animal enzymes:

[0109] (1) Animal source protein I (pork, chicken, and duck in a ratio of 1:1:1) was crushed and ground to about 140 mesh, mixed with water, and sterilized at 80°C for 2 h;

[0110] (2) Add sugar to the sterilized material in step (1) and stir evenly;

[0111] (3) Add the complex enzyme to the material treated in step (2), stir evenly, and perform enzymolysis at 37°C for 1 h;

[0112] (4) adjusting the pH value of the material after treatment in step (3) to 7.4-7.5, inoculating fermentation agent I, and performing aerobic fermentation at 37°C with stirring for 24 hours;

[0113] (5) adding fermentation agent II to the material after fermentation in step (4), and performing anaerobic fermentation at 30-37°C for 36 hours;

[0114] (6) The material after anaerobic fermentation in step (5) is crushed to 200 meshes to obtain animal enzyme 4.

[0115] Comparative Example 2: Replace Bacillus amyloliquefaciens with Bacillus licheniformis

[0116] The raw materials of the animal enzyme in this comparative example are as follows, in parts by weight, including:

[0117] Animal source protein I: 50 parts;

[0118] Animal source protein II includes: black soldier fly larvae: 15 parts, silkworm pupae: 15 parts;

[0119] Flaxseed: 6 parts; Honeysuckle: 6 parts; Viola yedoensis: 6 parts;

[0120] The compound enzyme includes: acid protease: 0.3 parts, lipase: 0.2 parts, cellulase: 0.2 parts;

[0121] The fermentation agent I comprises: 1 part of Bacillus licheniformis and 0.5 part of Bacillus subtilis;

[0122] The fermentation agent II includes: 1 part of Lactobacillus plantarum and 0.5 part of Lactobacillus acidophilus;

[0123] Sugar: 1.5 parts;

[0124] Water: 100 parts.

[0125] (1) Animal source protein I (pork, chicken, duck in a ratio of 1:1:1) and animal source protein II were crushed and ground to about 140 mesh; flaxseed, honeysuckle, and Viola yedoensis were crushed to 80 mesh, mixed with water, and sterilized at 80°C for 2 h;

[0126] (2) Add sugar to the sterilized material in step (1) and stir evenly;

[0127] (3) Add the complex enzyme to the material treated in step (2), stir evenly, and perform enzymolysis at 37°C for 1 h;

[0128] (4) adjusting the pH value of the material after treatment in step (3) to 6.5-7.0, inoculating fermentation agent I, and performing aerobic fermentation at 37°C with stirring for 24 h;

[0129] (5) adding fermentation agent II to the material after fermentation in step (4), and performing anaerobic fermentation at 30-37°C for 36 h;

[0130] (6) The material after anaerobic fermentation in step (5) is crushed to 200 meshes to obtain animal enzyme 5.

[0131] Test Example

[0132] Sources of cells and virus strains: Feline kidney fibroblast-like cells (F81) and feline calicivirus (FCV) were purchased from ATCC.

[0133] Reagents:

[0134] DMEM culture medium was purchased from Gibco; cck-8 was purchased from Bio-Tech;

[0135] Culture medium A: DMEM culture medium containing 10% FBS; Culture medium B: DMEM culture medium containing 2% FBS.

[0136] Test Example 1. In vitro safety test

[0137] Culture medium C containing animal enzymes: The animal enzymes obtained in the above examples and comparative examples are mixed with culture medium A in a volume ratio of 1:1, vortexed to mix, and then mixed with an equal volume of culture medium A after passing through a 0.22 μm membrane to obtain.

[0138] The F81 cells after passage were dispersed in culture medium A, inoculated in a 96-well plate, and cultured in a 37°C, 5% CO2 incubator; when the cells grew to 80% abundance, the culture medium was aspirated and then culture medium C containing animal enzymes or culture medium B without animal enzymes (as a control) (n=3) was added, and the cells were placed in a 37°C, 5% CO2 incubator for further culture for 48 h; after the culture medium was aspirated, the cells were washed twice with PBS, 180 μL of DMEM medium without FBS was added, and then 20 μL of cck-8 reagent was added, and the cells were incubated in a 37°C, 5% CO2 incubator for 1 h. The optical density (OD) at a wavelength of 450 nm was read using a microplate reader. The cell viability was calculated according to the following formula:

[0139] Cell viability = [OD (animal enzyme) - OD (blank)] / [OD (control) - OD (blank)] × 100%.

[0140] The results showed that the survival rate of cells treated with culture medium containing animal enzymes was greater than 95%, indicating its good safety.

[0141] Test Example 2. In vitro antiviral test

[0142] (1) TCID 50 )Determination:

[0143] The feline calicivirus FCV virus solution was diluted 10 times in a gradient to a final concentration of 10 -1 ~10 -10 / 100 μL;

[0144] The subcultured F81 cells were inoculated into 96-well plates and cultured in a 37°C, 5% CO2 incubator. When the cells grew to 80%, 100 μL of the above-mentioned virus solutions or virus-free culture medium were added to the 96-well plates (n=8), and 100 μL of culture medium B was added, and the plates were cultured in a 37°C, 5% CO2 incubator. The virus TCID was calculated according to the Reed-Muench method. 50 .

[0145] (2) Inactivation of FCV by animal enzymes

[0146] The animal enzymes obtained in the above examples and comparative examples were mixed with culture medium B at a volume ratio of 1:1, vortexed and mixed, and then passed through a 0.22 μm membrane and then mixed with an equal volume of culture medium B containing feline calicivirus FCV virus (containing 100 TCID 50 The FCV of 1% FCV was mixed and treated at 37°C for 120 min, and then the supernatant was removed by ultracentrifugation. The precipitate was washed with PBS and then ultracentrifuged again to precipitate the virus particles. The virus particles were resuspended in nutrient solution B and then inoculated into a 96-well plate (with a cell abundance of about 80%) inoculated with F81 cells after passage (n=3). At the same time, a cell control group without any treatment of the cells and a virus control group treated with the virus only were set up. The cells were cultured for 72 h. The virus inhibition rate was calculated using CCK-8 according to the method in Test Example 1. The calculation method is as follows:

[0147] Virus inhibition rate = [OD (animal enzyme) - OD (virus control)] / [OD (cell control) - OD (virus control)] × 100%.

[0148] The results are shown in Table 1.

[0149] Table 1

[0150] FCV virus inhibition rate (%) Animal enzyme 1a 6.1±2.4 Animal enzyme 1b 15.3±4.1 Animal Enzyme 1c 19.2±2.1 Animal enzyme 1d 9.3±3.3 Animal Enzyme 1e 5.5±0.8 Animal enzyme 1f 2.4±1.9 Animal Enzyme 2 16.9±1.5 Animal Enzyme 3 15.9±3.0 Animal Enzyme 4 0.1±1.9* Animal Enzyme 5 3.1±2.2*

[0151] * indicates that there is a significant difference in virus inhibition rate compared with the aerobic fermentation 24 h group in Example 1, P < 0.05.

[0152] From the data in Table 1, it can be seen that the animal enzyme prepared in the embodiment of the present invention has a certain direct inactivation effect on the feline calicivirus FCV virus. In particular, the inhibitory effect of the animal enzymes prepared with different aerobic fermentation times on the feline calicivirus FCV virus in Example 1 is very different. The aerobic fermentation time with better effect is 20~28 h. The animal enzymes with too short or too long aerobic fermentation have poor inhibitory effects on the virus. The inventor analyzed the reasons and found that the short fermentation time may make it impossible to release effective antiviral substances or the effective substances produced by microbial metabolism have not been synthesized; and too long fermentation time leads to the destruction of effective ingredients (such as peptides, polysaccharides, etc.). Therefore, it is necessary to strictly control the aerobic fermentation time to ensure the effectiveness of animal enzymes in antiviral.

[0153] From the data of the animal enzyme virus inhibition effect obtained in Comparative Examples 1 and 2, it is significantly different from the examples, and the direct inactivation effect on the feline calicivirus FCV virus is almost non-existent or extremely slight. This shows that the black soldier fly larvae and silkworm pupae as animal proteins and the Chinese medicinal ingredients linseed, honeysuckle and Viola yedoensis play a key role in the fermentation system against the feline calicivirus FCV virus. In addition, the selection of strains is another key factor. Even if only Bacillus amyloliquefaciens is replaced with Bacillus licheniformis, it cannot produce an anti-feline calicivirus FCV virus effect. This may be caused by differences in the metabolic pathways and metabolites of different strains.

[0154] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An animal enzyme, characterized in that: Made from the following raw materials in parts by weight: Animal source protein I: 40~60 parts; Animal source protein II includes: black soldier fly larvae: 10-20 parts, silkworm pupae: 10-20 parts; Flaxseed: 4-8 parts; Honeysuckle: 4-8 parts; Viola yedoensis: 4-8 parts; The compound enzyme includes: acid protease: 0.2-0.6 parts, lipase: 0.1-0.3 parts, cellulase: 0.1-0.3 parts; The fermentation agent I comprises: 0.5-2 parts of Bacillus amyloliquefaciens and 0.5-0.2-1 parts of Bacillus subtilis; The fermentation agent II includes: 0.5-2 parts of Lactobacillus plantarum 1 and 0.5-0.2-1 parts of Lactobacillus acidophilus; Sugar: 1~3 parts; Water: 80~120 parts; Wherein, the animal source protein I is one or more of livestock meat or poultry meat; The method for preparing the animal enzyme comprises the following steps: (1) Mix the ground flaxseed, honeysuckle, Viola yedoensis, and the ground animal protein I and animal protein II with water, and then sterilize them; (2) Add sugar to the sterilized material in step (1) and mix well; (3) Adding the complex enzyme to the material treated in step (2), mixing well, and performing enzymolysis at 30-37°C for 0.5-2 h; (4) adjusting the pH of the material treated in step (3) to 7.0-8.0, inoculating fermentation agent I, and aerobically fermenting at 30-37°C for 20-28 h; (5) Add fermentation agent II to the material after fermentation in step (4) and perform anaerobic fermentation at 30-37°C for 24-48 hours.

2. The animal enzyme according to claim 1, characterized in that The animal enzyme is made from the following raw materials in parts by weight: Animal source protein I: 45-55 parts; Animal source protein II includes: black soldier fly larvae: 12-18 parts, silkworm pupae: 12-18 parts; Flaxseed: 5-7 parts; Honeysuckle: 5-7 parts; Viola yedoensis: 5-7 parts; The compound enzyme includes: acid protease: 0.2-0.4 parts, lipase: 0.1-0.3 parts, cellulase: 0.1-0.3 parts; The fermentation agent I comprises: 0.8-1.2 parts of Bacillus amyloliquefaciens and 0.3-0.8 parts of Bacillus subtilis; The fermentation agent II includes: 0.8-1.2 parts of Lactobacillus plantarum and 0.3-0.8 parts of Lactobacillus acidophilus; Sugar: 1~2 parts; Water: 90~110 parts.

3. The animal enzyme according to claim 1, characterized in that The sugar is selected from glucose, starch or a combination thereof.

4. The preparation method according to claim 1, characterized in that: In step (1), the sterilization temperature is 80-100°C and the time is 1-4 h.

5. The preparation method according to claim 1, characterized in that: In step (3), the enzymatic hydrolysis temperature is 35-37°C.

6. The preparation method according to claim 1, characterized in that: In step (4), the pH of the material treated in step (3) is adjusted to 7.4-7.

5.

7. The preparation method according to claim 1, characterized in that: In step (4), the temperature of aerobic fermentation is 35-37°C, and the fermentation time is 20-28 h.

8. The preparation method according to claim 1, characterized in that: In step (5), the temperature of anaerobic fermentation is 35-37°C and the fermentation time is 24-36 h.

9. Use of the animal enzyme according to any one of claims 1 to 8 in the preparation of a drug against feline calicivirus.