Enzyme preparation for degrading allergen of pet food and application of enzyme preparation

Abstract

The invention discloses an enzyme preparation for degrading allergens of pet food and application of the enzyme preparation, the enzyme preparation is a pH-responsive acrylic resin-pectin composite double-response shell embedded enzyme-probiotic composite system, a core layer is heat-resistant neutral protease, and a middle layer is a compound of lactobacillus reuteri and bacillus subtilis. In the processing stage, the protease degrades macromolecular allergens, the composite shell in the gastrointestinal environment firstly triggers outer acrylic resin to be dissolved through weak alkalinity of the intestinal tract, then inner pectin is degraded by pancreatin, and probiotics are accurately released to metabolize residual sensitizing peptides. The enzyme preparation realizes full-chain desensitization, is high in survival rate of probiotics, is suitable for various pet foods, and is stable and reliable in desensitization effect.

Description

Technical Field

[0001] This invention relates to the field of pet food additives, specifically to an enzyme preparation for degrading pet food allergens and its application. Background Technology

[0002] With the development of the pet food industry, protein allergens in raw materials such as meat and grains easily trigger allergic reactions in pets, such as skin itching and digestive discomfort, becoming a key issue affecting pet health. Current technologies mostly use single enzyme preparations to degrade allergens, but common proteases have poor thermostability and are easily inactivated during the high-temperature extrusion process of pet food at 80-100℃. Furthermore, they lack a targeted release mechanism and are prone to premature degradation and inactivation in the stomach environment. Some solutions add probiotics to assist in intestinal regulation, but the survival rate of probiotics is low, making it difficult to form a synergistic effect with enzyme preparations. In addition, commonly used encapsulation materials are mostly pH-responsive, unable to accurately match the pH fluctuations of the pet's intestines (5.5-7.5), and the degradation rate of the encapsulation layer does not match the release rhythm of probiotics and enzyme preparations, resulting in limited allergen degradation efficiency. Therefore, developing an allergen-degrading enzyme preparation that combines high-temperature stability, targeted release characteristics, and the ability to achieve synergistic effects between enzymes and bacteria, to solve the problems of easy enzyme inactivation, inaccurate release, and poor synergistic effects in existing technologies, has become an urgent need in the field of pet food additives. Summary of the Invention

[0003] To address the aforementioned problems, this invention proposes an enzyme preparation for degrading pet food allergens and its application. This enzyme preparation adopts a layered structure design consisting of a core enzyme layer, a probiotic layer, and a composite outer shell layer. It combines thermal stability with intestinal targeted release characteristics and exhibits excellent performance in terms of allergen degradation efficiency, probiotic survival rate, and compatibility with the pet's intestinal environment. It can meet the application needs of high-temperature processing of pet food and prevention and control of pet intestinal allergies.

[0004] To achieve the above objectives, the present invention adopts the following technical solution: An enzyme preparation for degrading pet food allergens, characterized in that the enzyme preparation is prepared from the following raw materials in parts by weight, and the enzyme preparation comprises, from the inside out, a core enzyme layer, a probiotic layer, and a composite outer shell layer: Neutral protease 5-10 parts, Lactobacillus reuteri 3-6 parts, Bacillus subtilis 2-4 parts, pH-responsive acrylic resin 15-25 parts, pectin 8-12 parts, polyethylene glycol 6000 1.2-2.0 parts, food-grade carrier 1-3 parts, protectant 0.5-1.5 parts; The pH-responsive acrylic resin swells and dissolves in an environment with a pH of 5.5-7.5; the pectin can be specifically degraded by intestinal pancreatic enzymes, and the pectin is methoxylated pectin; the polyethylene glycol 6000 is a plasticizer for the pH-responsive acrylic resin; the food-grade carrier is selected from one or more of maltodextrin, lactose, mannitol, and corn starch, and the carrier is only used for core enzyme layer formation; the protectant is selected from one or more of skim milk powder, trehalose, and glycerol, and the protectant is only used for live bacteria protection of the probiotic layer.

[0005] Optionally, the neutral protease is a heat-stable neutral protease with an enzyme activity of 5000-8000 U / g, maintaining stable enzyme activity at high temperatures of 80-100℃.

[0006] Optionally, the heat-stable neutral protease is selected from one or more of Streptococcus thermophilus, Bacillus subtilis, or Aspergillus oryzae.

[0007] Optionally, the pH-responsive acrylic resin is selected from one or more of methacrylic acid-methyl methacrylate copolymer, methacrylic acid-ethyl acrylate copolymer, and methacrylic acid-propyl acrylate copolymer, and the monomer molar ratio of the copolymer is methacrylic acid: acrylate monomer = 1:1-2:1.

[0008] Optionally, the probiotic layer contains 1 × 10⁻⁶ live Lactobacillus reuteri. 10 -5×10 10 CFU / g, viable count of Bacillus subtilis is 5 × 10⁻⁶. 9 -2×10 10 CFU / g.

[0009] Optionally, in the composite outer shell layer, the outer layer of pH-responsive acrylic resin has a thickness of 50-70 μm, and the inner layer of pectin has a thickness of 30-50 μm.

[0010] Optionally, the pectin is selected from one or two of low-methoxyl pectin and high-methoxyl pectin, wherein the methoxyl content of low-methoxyl pectin is 2%-15% and the methoxyl content of high-methoxyl pectin is 15%-30%.

[0011] Optionally, the preparation method of the enzyme preparation for degrading pet food allergens includes the following specific preparation steps: S1. Neutral protease and food-grade carrier are mixed in parts by weight, and an appropriate amount of deionized water is added to adjust the slurry with a solid content of 30%-40%. The slurry is then spray-dried and shaped with an inlet air temperature of 120-140℃, an outlet air temperature of 60-80℃, and an atomization pressure of 0.3-0.5 MPa to obtain core enzyme particles with a particle size of 100-200 μm. S2. Mix Lactobacillus reuteri, Bacillus subtilis and protectant, add deionized water to make a bacterial suspension with a solid content of 15%-20%, and use fluidized bed coating method, with inlet air temperature of 40-50℃ and spray rate of 5-10 mL / min to coat the core enzyme particles to form a probiotic layer with a thickness of 20-30 μm. S3. Dissolve pH-responsive acrylic resin, pectin, and polyethylene glycol 6000 in an ethanol-water solution at a volume ratio of 3:7-5:5 to prepare a coating solution with a mass concentration of 10%-15%. Use a fluidized bed coating method with an inlet air temperature of 50-60℃ and a spraying rate of 8-15 mL / min to sequentially coat the surface of the probiotic layer. After drying at a constant temperature of 45-55℃ for 30-60 min, the enzyme preparation is obtained.

[0012] Optionally, the application of the enzyme preparation for degrading pet food allergens, wherein the pet food includes puffed food, wet food and snacks, and the enzyme preparation is added to the pet food at a rate of 0.1%-0.5%.

[0013] The beneficial effects of this invention are as follows: The enzyme preparation prepared by this invention for degrading pet food allergens has a degradation rate of over 90% for common allergens such as chicken ovalbumin and zein, which can significantly reduce the risk of pet allergies; the heat-stable neutral protease retains over 85% of its activity during high-temperature processing at 80-100℃, solving the problem of easy inactivation of conventional proteases. The three-layer structure design enables the composite outer shell layer to achieve intestinal-targeted release, increasing the survival rate of probiotics to over 90%, forming a synergistic anti-allergy effect with the protease; the dense coating structure also extends the shelf life of the enzyme preparation, and the food-grade raw materials ensure biosafety, making it suitable for the production and processing needs of various pet foods. Attached Figure Description

[0014] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof.

[0015] Figure 1 This is a bar chart comparing the allergen degradation rates of different samples in this invention. Figure 2 This is a bar chart comparing the targeted release rates of enzymes in different samples of this invention under gastric / intestinal conditions. Detailed Implementation

[0016] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0017] Example 1: This Example 1 describes an enzyme preparation for degrading pet food allergens. The enzyme preparation is prepared from the following raw materials in parts by weight: 7 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 20 parts pH-responsive acrylic resin: methyl methacrylate copolymer (Eudragit L100), 10 parts pectin: low methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier: maltodextrin, 1.0 part trehalose as a preservative; This embodiment describes a method for preparing an enzyme preparation that degrades pet food allergens. The specific preparation steps are as follows: S1. Neutral protease and food-grade carrier are mixed in parts by weight, and an appropriate amount of deionized water is added to make a slurry with a solid content of 35%. The slurry is then spray-dried and shaped with an inlet air temperature of 130°C, an outlet air temperature of 70°C, and an atomization pressure of 0.4 MPa to obtain core enzyme particles with a particle size of 150 μm. S2. Mix Lactobacillus reuteri, Bacillus subtilis and a protectant, add deionized water to make a bacterial suspension with a solid content of 18%, and use a fluidized bed coating method with an inlet air temperature of 45℃ and a spraying rate of 8 mL / min to coat the core enzyme particles to form a probiotic layer with a thickness of 25 μm. S3. Dissolve pH-responsive acrylic resin, pectin, and polyethylene glycol 6000 in an ethanol-water solution at a volume ratio of 4:6 to prepare a 12% (w / w) coating solution. Use a fluidized bed coating method with an inlet air temperature of 55°C and a spray rate of 12 mL / min to coat the surface of the probiotic layer. After drying at a constant temperature of 50°C for 45 min, the enzyme preparation is obtained.

[0018] Example 2: This embodiment 2 presents an enzyme preparation for degrading pet food allergens. The enzyme preparation is prepared from the following raw materials in parts by weight: 5 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 20 parts pH-responsive acrylic resin: methyl methacrylate copolymer (Eudragit L100), 10 parts low-methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier: maltodextrin, 1.0 part trehalose as a preservative; In this embodiment, the preparation method of an enzyme preparation for degrading pet food allergens is the same as in Example 1, except that the amount of neutral protease is reduced to 5 parts.

[0019] Example 3: This embodiment 3 describes an enzyme preparation for degrading pet food allergens. The enzyme preparation is prepared from the following raw materials in parts by weight: 10 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 20 parts pH-responsive acrylic resin: methyl methacrylate copolymer (Eudragit L100), 10 parts low-methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier: maltodextrin, 1.0 part trehalose as a preservative; In this embodiment, the preparation method of an enzyme preparation for degrading pet food allergens is the same as in Example 1, except that the amount of neutral protease is increased to 10 parts.

[0020] Comparative Example 1: The enzyme preparation in Comparative Example 1 was prepared from the following parts by weight of raw materials: 7 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 10 parts low-methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier maltodextrin, and 1.0 part trehalose as a preservative. The enzyme preparation method in this comparative example is the same as in Example 1, except that pH-responsive acrylic resin is not added.

[0021] Comparative Example 2: The enzyme preparation in Comparative Example 2 was prepared from the following parts by weight of raw materials: 7 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 20 parts pH-responsive acrylic resin: methyl methacrylate copolymer (Eudragit L100), 10 parts pectin: low methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier: maltodextrin, 1.0 part trehalose as a preservative; The specific preparation steps of the enzyme preparation in this comparative example are as follows: S1. Add neutral protease, Lactobacillus reuteri, Bacillus subtilis, pH-responsive acrylic resin, pectin, polyethylene glycol 6000, food-grade carrier, and protectant into a mixer and stir for 15 minutes to obtain a uniform mixed powder. S2. Add deionized water to the mixed powder to make a slurry with a solid content of 35%; S3. The slurry is fed into a spray drying device and spray dried to form granules. The inlet air temperature is 130℃, the outlet air temperature is 70℃, and the atomization pressure is 0.4 MPa to obtain mixed granules.

[0022] Comparative Example 3: The enzyme preparation in Comparative Example 3 was prepared from the following parts by weight of raw materials: 7 parts neutral protease, 4 parts Lactobacillus reuteri, 3 parts Bacillus subtilis, 20 parts pH-responsive acrylic resin: methyl methacrylate copolymer (Eudragit L100), 10 parts pectin: low methoxyl pectin, 1.5 parts polyethylene glycol 6000, 2 parts food-grade carrier: maltodextrin, 1.0 part trehalose as a preservative; The preparation method of the enzyme preparation in this comparative example is the same as that in Example 1, except that a heat-insensitive neutral protease is used.

[0023] Performance testing 1. Allergen degradation rate test High-performance liquid chromatography (HPLC) was used to prepare a simulated pet food matrix containing chicken oocyte mucoprotein, with an allergen concentration of 3 mg / g. Enzyme preparation was added to the simulated matrix at the application ratio (0.3%) of this invention, and the matrix was incubated at 37°C in a shaker for 3 h to simulate the intestinal digestive environment of a pet. After incubation, an appropriate amount of hydrochloric acid was added to the system to terminate the enzyme reaction. The supernatant was collected by centrifugation (8000 rpm, 10 min), filtered through a 0.45 μm filter membrane, and the residual allergen content in the supernatant was detected by HPLC (column: C18, mobile phase: methanol-water = 40:60, flow rate 1.0 mL / min, detection wavelength 280 nm). A blank control group without added enzyme preparation was also set up, treated and tested using the same steps. Finally, the degradation rate of the target allergen by the enzyme preparation was calculated using the formula: "Degradation rate = (Allergen content in the blank control group - Residual allergen content in the experimental group) / Allergen content in the blank control group × 100%".

[0024] Table 1. Test data on allergen degradation rate of different samples The enzyme preparations in Examples 1-3 all showed a degradation rate of over 90% for chicken oocyte mucoprotein, with Example 3 reaching 93.7%, which was significantly better than the comparative examples (62.7%-78.7%). This indicates that the enzyme preparations of the present invention can efficiently degrade pet food allergens and meet the core function requirements.

[0025] 2. High-temperature enzyme activity retention rate test Referring to GB / T 23527-2009 "Protein Preparations", the Folin-phenol method was adopted. First, enzyme preparation samples from each example and comparative example were accurately weighed and dissolved in buffer solution to determine the initial enzyme activity (denoted as A0). Then, simulating the high-temperature extrusion process of pet food, the enzyme preparation samples were placed in a 90℃ constant temperature drying oven for 30 min. After cooling to room temperature, the enzyme activity was determined again under the same concentration and detection conditions (denoted as A1). At the same time, a blank control group (buffer solution only, no enzyme preparation) was set up to eliminate interference. Finally, the enzyme activity retention rate of each sample was calculated according to the formula "high-temperature enzyme activity retention rate = (A1 / A0) × 100%". Three parallel experiments were set up for each sample, and the average value was taken as the final result.

[0026] Table 2. Data on enzyme activity retention rate at high temperatures for different samples After being treated at 90°C, Examples 1-3 showed an enzyme activity retention rate of over 86%, while Comparative Example 3 (ordinary neutral protease) only had a retention rate of 42.0%. This demonstrates that the thermostable neutral protease binding layered structure of the present invention can effectively withstand the high-temperature processing of pet food and solve the problem of easy inactivation of conventional enzymes.

[0027] 3. Probiotic intestinal survival rate test Accurately weigh the enzyme preparation samples of each example and comparative example, and determine the initial total viable count using a serial dilution method (Lactobacillus reuteri was cultured on MRS medium, Bacillus subtilis on LB medium, and counted after incubation at 37°C for 48 h, denoted as B0). Subsequently, simulating the gastrointestinal digestive environment of a pet, the samples were added to artificial gastric fluid (pH 1.5, containing pepsin) and incubated at 37°C on a shaker for 2 h, and then transferred to artificial intestinal fluid (pH 7.0, containing trypsin) for further incubation for 4 h. After incubation, the number of surviving viable bacteria was determined according to the same dilution factor and culture conditions (denoted as B1). At the same time, a sterile control group was set up to eliminate contamination interference. Finally, the survival rate of each sample was calculated according to the formula "Probiotic intestinal survival rate = (B1 / B0) × 100%". Three parallel experiments were set up for each sample, and the average value was taken as the final result.

[0028] Table 3. Data on intestinal survival rate of probiotics in different samples The intestinal survival rate of probiotics in Examples 1-3 exceeded 89%, with Example 1 reaching 91.2%, while Comparative Example 2 (non-layered structure) only reached 68.7%. This confirms that the layered structure and composite shell of the present invention can effectively protect probiotics as they pass through the gastrointestinal tract and ensure their survival efficacy.

[0029] 4. Enzyme targeted release test Accurately weigh the enzyme preparation samples from each example and comparative example, and determine the total enzyme activity (denoted as T) using the Folin-phenol method. Then, simulating a two-step digestive environment in a pet's gastrointestinal tract, the sample was first added to artificial gastric fluid (pH 1.5, containing pepsin, but without pancreatin) preheated to 37°C and incubated on a constant-temperature shaker (100 rpm) for 2 hours. After incubation, the supernatant was collected by centrifugation (8000 rpm, 10 min), and the enzyme activity released in the gastric environment was determined (denoted as G). Then, an equal volume of artificial intestinal fluid (pH 7.0, containing pancreatin, but without pepsin) was added to the remaining precipitate, and incubation was continued at 37°C and 100 rpm for 4 hours. h, the supernatant was centrifuged and the enzyme activity released in the intestinal environment was measured (denoted as I); at the same time, a blank control group (artificial gastric juice / intestinal juice only, without enzyme preparation) was set up to eliminate interference. Finally, the release performance index was calculated according to the formula "gastric environment release rate = (G / T) × 100%" and "intestinal environment total release rate = (G+I) / T × 100%". Three parallel experiments were set up for each sample, and the average value was taken as the final result.

[0030] Table 4. Targeted release test data of enzymes from different samples In Examples 1-3, the gastric environment release rate was only 7.0%, while the total intestinal environment release rate exceeded 94%. Comparative Example 1 (lacking acrylic resin) and Comparative Example 2 (non-layered structure) showed significantly higher gastric environment release rates, indicating that the composite outer shell layer achieved a targeted design of "stable gastric and intestinal release," ensuring that the enzymes functioned efficiently in the intestine.

[0031] 5. Acute toxicity test Referring to GB 15193.3-2014, 40 healthy ICR mice (half male and half female, weighing 18-22 g) were randomly divided into an experimental group (Example 1) and a blank control group (physiological saline), with 20 mice in each group. The experimental group received a high dose of 5000 mg / kg body weight, with the enzyme preparation of this invention prepared as a suspension in physiological saline and administered orally by gavage at a volume of 0.2 mL / 10 g body weight. The blank control group received only the same volume of physiological saline by gavage. After administration, the mice were placed in a specific pathogen-free (SPF) animal room (temperature 22-25℃, relative humidity 55%-65%, 12 h light / 12 h dark cycle) for routine feeding and watering, and were observed continuously for 14 days. Daily records were made of the mice's appearance, behavior, food and water intake, and mortality. After the experiment, the mice's weight was measured, and surviving mice were dissected to observe any abnormal lesions in the major organs (heart, liver, spleen, lungs, and kidneys). The median lethal dose (LD50) was determined based on the number of mice that died. 50 ), to assess the acute toxicity of enzyme preparations.

[0032] Table 5. Acute toxicity test data for different samples Mice in the experimental group, after being administered a dose of 5000 mg / kg body weight via gavage, showed no death or symptoms of poisoning within 14 days, exhibited normal weight gain, and showed no organ abnormalities. The LD50 was determined accordingly. 50 ≥5000 mg / kg body weight, which is practically non-toxic and meets the safety requirements for pet food additives.

[0033] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An enzyme preparation for degrading pet food allergens, characterized in that, The enzyme preparation is prepared from the following raw materials in parts by weight, and the enzyme preparation consists of a core enzyme layer, a probiotic layer, and a composite outer shell layer from the inside out: Neutral protease 5-10 parts, Lactobacillus reuteri 3-6 parts, Bacillus subtilis 2-4 parts, pH-responsive acrylic resin 15-25 parts, pectin 8-12 parts, polyethylene glycol 6000 1.2-2.0 parts, food-grade carrier 1-3 parts, protectant 0.5-1.5 parts; The pH-responsive acrylic resin swells and dissolves in an environment with a pH of 5.5-7.5; the pectin can be specifically degraded by intestinal pancreatic enzymes, and the pectin is methoxylated pectin; the polyethylene glycol 6000 is a plasticizer for the pH-responsive acrylic resin; the food-grade carrier is selected from one or more of maltodextrin, lactose, mannitol, and corn starch, and the carrier is only used for core enzyme layer formation; the protectant is selected from one or more of skim milk powder, trehalose, and glycerol, and the protectant is only used for live bacteria protection of the probiotic layer.

2. The enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, The neutral protease is a thermostable neutral protease with an enzyme activity of 5000-8000 U / g, and its enzyme activity remains stable at high temperatures of 80-100℃.

3. The enzyme preparation for degrading pet food allergens according to claim 2, characterized in that, The heat-stable neutral protease is selected from one or more of Streptococcus thermophilus, Bacillus subtilis, or Aspergillus oryzae.

4. The enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, The pH-responsive acrylic resin is selected from one or more of methacrylic acid-methyl methacrylate copolymer, methacrylic acid-ethyl acrylate copolymer, and methacrylic acid-propyl acrylate copolymer, and the monomer molar ratio of the copolymer is methacrylic acid: acrylate monomer = 1:1-2:

1.

5. The enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, The probiotic layer contains 1 × 10⁻⁶ live Lactobacillus reuteri. 10 -5×10 10 CFU / g, viable count of Bacillus subtilis is 5 × 10⁻⁶. 9 -2×10 10 CFU / g.

6. The enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, In the composite outer shell layer, the outer layer of pH-responsive acrylic resin has a thickness of 50-70 μm, and the inner layer of pectin has a thickness of 30-50 μm.

7. The enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, The pectin is selected from one or two of low-methoxyl pectin and high-methoxyl pectin, wherein the methoxyl content of low-methoxyl pectin is 2%-15% and the methoxyl content of high-methoxyl pectin is 15%-30%.

8. The method for preparing an enzyme preparation for degrading pet food allergens according to claim 1, characterized in that, The specific preparation steps are as follows: S1. Neutral protease and food-grade carrier are mixed in parts by weight, and an appropriate amount of deionized water is added to adjust the slurry with a solid content of 30%-40%. The slurry is then spray-dried and shaped with an inlet air temperature of 120-140℃, an outlet air temperature of 60-80℃, and an atomization pressure of 0.3-0.5 MPa to obtain core enzyme particles with a particle size of 100-200 μm. S2. Mix Lactobacillus reuteri, Bacillus subtilis and protectant, add deionized water to make a bacterial suspension with a solid content of 15%-20%, and use fluidized bed coating method, with inlet air temperature of 40-50℃ and spray rate of 5-10 mL / min to coat the core enzyme particles to form a probiotic layer with a thickness of 20-30 μm. S3. Dissolve pH-responsive acrylic resin, pectin, and polyethylene glycol 6000 in an ethanol-water solution at a volume ratio of 3:7-5:5 to prepare a coating solution with a mass concentration of 10%-15%. Use a fluidized bed coating method with an inlet air temperature of 50-60℃ and a spraying rate of 8-15mL / min to sequentially coat the surface of the probiotic layer. After drying at a constant temperature of 45-55℃ for 30-60 min, the enzyme preparation is obtained.

9. The application of an enzyme preparation for degrading pet food allergens according to any one of claims 1-7, characterized in that, The pet food includes puffed food, wet food, and snacks, and the enzyme preparation is added to the pet food at a rate of 0.1%-0.5%.

Images