Fermentation preparation for feed production process and preparation technology thereof

Abstract

The invention relates to the technical field of animal feed fermentation, in particular to a fermentation preparation in a feed production process and a preparation technology of the fermentation preparation. The fermentation preparation is composed of microcapsules, the microcapsules comprise a core material layer and a coating wall material layer, the core material layer comprises lactobacillus plantarum, candida utilis, betaine and peptone, the coating wall material layer comprises an inner layer and an outer layer, the inner layer is a (trehalose-6-phosphoric acid)-beeswax compound, and the outer layer is a (trehalose-6-phosphoric acid)-beeswax compound. And the outer layer is a monoglyceride citrate-Arabic gum compound. After entering a feed production environment, the fermentation preparation is subjected to three-stage change: firstly, an outer layer glyceride citrate-Arabic gum compound is dissolved at proper temperature and humidity to form a network channel; secondly, a transition area is formed between the inner layer and the outer layer through intermolecular interaction, and permeation is adjusted; finally, the inner layer (trehalose-6-phosphoric acid)-beewax compound forms an electrostatic network, so that the protective release of active substances is realized, and the fermentation quality is improved.

Description

Technical Field

[0001] The invention relates to the technical field of animal feed fermentation, and in particular to a fermentation preparation in a feed production process and a preparation process thereof. Background Art

[0002] At present, the application of fermentation preparations in the feed production process has been quite extensive and is becoming more and more mature. The main fermentation preparations used include microbial preparations such as yeast, lactic acid bacteria, and Bacillus, which can significantly improve the quality of feed, improve the digestion and absorption capacity of animals, and promote growth. In actual production, the use of fermentation preparations is mainly concentrated in the following aspects: first, in the fermentation process of feed raw materials, by adding special fermentation strains to degrade the anti-nutritional factors in the raw materials, improve the utilization rate of nutrients such as protein, and produce beneficial metabolites such as organic acids, vitamins and enzymes; second, directly add probiotic preparations to compound feed. These active strains can colonize and reproduce after entering the animal's digestive tract, regulate the balance of intestinal flora, and enhance immunity; third, use fermentation preparations to process agricultural and sideline products and industrial waste materials and convert them into high-quality protein feed.

[0003] At present, there are still some technical problems to be solved in the application of fermentation preparations in the feed industry. First, in feed processing projects, the survival rate of active strains is generally low, which is mainly due to the inability of traditional preparations to effectively resist the influence of high temperature, high pressure and mechanical shear force in the process of granulation and puffing. Secondly, the storage stability of existing fermentation preparations is insufficient. During transportation and storage, the number of active strains will drop significantly due to environmental factors such as temperature, humidity, and oxygen, resulting in a rapid decrease in product potency. In addition, most of the fermentation preparations on the market are based on a single strain, and there is a lack of products with synergistic effects of multiple strains, which limits the functionality and application effect of fermentation preparations. Summary of the invention

[0004] (1) Technical issues to be resolved The purpose of the present invention is to provide a fermentation preparation in a feed production process and a preparation process thereof, so as to solve the problems that the survival rate of active strains in the feed processing process is low and the storage stability of existing fermentation preparations is insufficient.

[0005] (2) Technical solution To achieve the above-mentioned purpose, on the one hand, the present invention provides a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0006] Furthermore, a fermentation preparation for a feed production process, the core material layer comprises the following raw materials in parts by weight: 15-25 parts of Lactobacillus plantarum, 20-30 parts of Candida utilis, 8-12 parts of betaine, and 10-15 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 25-35 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 15-25 parts of glyceryl citrate-gum arabic complex.

[0007] Furthermore, the preparation method of the (trehalose-6-phosphate)-beeswax complex comprises: S11. The beeswax is heated to 65-70 ° C to obtain melted beeswax, the melted beeswax is added to a phosphate buffer at 65-70 ° C, and then stirred at 400-500 rpm for 30-40 min to obtain a beeswax emulsion; S12. adding trehalose-6-phosphate to phosphate buffer to obtain a trehalose-6-phosphate solution, adding the trehalose-6-phosphate solution dropwise to the beeswax emulsion, and reacting at 300-400 rpm for 2-3 hours to obtain a reactant; S13. Centrifuge the reactant at 3000-4000 rpm for 15-20 min to obtain a precipitate, wash the precipitate with deionized water for 3-4 times to obtain a washed product, vacuum dry the washed product at 40-50° C. for 12-24 h to obtain a dried product, grind the dried product into powder to obtain a (trehalose-6-phosphate)-beeswax complex.

[0008] Furthermore, the preparation method of the glycerol citrate-gum arabic complex comprises: S21. Adding gum arabic to deionized water and stirring at 40-50° C. for 2-3 h to obtain a gum arabic solution; S22. Adding glyceryl citrate to anhydrous ethanol to obtain glyceryl citrate solution; S23. Slowly add the glycerol citrate solution to the gum arabic solution at a dropping speed of 2 to 3 drops / s. After the dropping is completed, stir at 300 to 400 rpm for 4 to 5 hours to obtain a reactant; S24. The reactant is ultrasonically treated for 15 to 20 minutes to obtain an ultrasonic reactant, and the ultrasonic reactant is centrifuged at 4000 to 5000 rpm for 20 to 25 minutes to obtain a centrifuged reactant; S25. Wash the centrifuged reactant 3 to 5 times with a mixed solvent consisting of water and ethanol in a volume ratio of 1:1 to obtain a washed reactant, freeze-dry the washed reactant for 36 to 48 hours to obtain a dried reactant, grind the dried reactant into powder to obtain a glyceryl citrate-gum arabic complex.

[0009] On the other hand, based on the same inventive concept, the present invention also provides a preparation process of a fermentation preparation in a feed production process, which is applied to the fermentation preparation in the feed production process, comprising the following steps: S31. The Lactobacillus plantarum was activated and cultured in MRS medium at 37°C for 24 hours to obtain the cultured Lactobacillus plantarum, the cultured Lactobacillus plantarum was centrifuged at 4000-5000 rpm for 10-15 minutes to obtain Lactobacillus plantarum, and the Lactobacillus plantarum was washed 2-3 times with phosphate buffer to obtain the activated Lactobacillus plantarum; S32. The utilis strain was activated and cultured in YPD medium at 28°C for 48h to obtain cultured utilis Candida, the cultured utilis Candida was centrifuged at 3500-4000rpm for 15-20min to obtain utilis Candida cells, and the utilis Candida cells were washed 2-3 times with phosphate buffer to obtain activated utilis Candida; S33. The activated Lactobacillus plantarum and Candida utilis cells were mixed to obtain a mixed cell, the mixed cell was added to a betaine solution to obtain a first mixed solution, the first mixed solution was added with peptone to obtain a second mixed solution, the second mixed solution was stirred at 300 to 400 rpm for 30 to 40 min to obtain a stirred mixed solution, the pH of the stirred mixed solution was adjusted to 6.8 to 7.2 to obtain a core liquid; S34. The prepared core liquid was spray dried to obtain initial particles at an air inlet temperature of 45 to 50°C, an air outlet temperature of 35 to 40°C, and an atomization pressure of 0.2 MPa; S35. Add (trehalose-6-phosphate)-beeswax complex to deionized water and stir at 10000-15000 rpm for 5-10 min to obtain (trehalose-6-phosphate)-beeswax complex dispersion, place core particles in a fluidized bed, and spray the (trehalose-6-phosphate)-beeswax complex dispersion from above at a spray pressure of 0.15-0.20 MPa, an inlet air temperature of 35-40° C., a bed temperature of 30-35° C., an atomization wind speed of 35-45 m / s, and a spray rate of 8-12 mL / min to obtain an inner coating structure; S36. Add the glyceryl citrate-gum arabic complex to deionized water and ultrasonically disperse it for 15 to 20 minutes to obtain a glyceryl citrate-gum arabic complex dispersion, place the inner coating structure in a fluidized bed, and spray the glyceryl citrate-gum arabic complex dispersion from above at a spray pressure of 0.12 to 0.15 MPa, an air inlet temperature of 40 to 45° C., a bed temperature of 35 to 38° C., an atomization wind speed of 30 to 40 m / s, and a spray rate of 6 to 10 mL / min to obtain an outer coating structure; S37. The outer coating structure is vacuum dried at 35-40°C for 4-6 hours to obtain a dried product, and the dried product is sieved through an 80-mesh sieve to obtain a fermentation preparation.

[0010] The mechanism of action of the above raw material components is as follows: Lactobacillus plantarum: It inhibits the growth of harmful bacteria by producing active substances such as organic acids and bacteriocins. It can also secrete a variety of digestive enzymes to promote the decomposition of feed nutrients, improve feed utilization, regulate the balance of animal intestinal flora, and enhance immunity.

[0011] Candida utilis: It can secrete active substances such as β-glucan and manno-oligosaccharide, has good antioxidant and immunomodulatory functions, can promote animal growth and development, improve feed conversion rate, and can also secrete a variety of vitamins and bioactive substances to improve feed quality.

[0012] Betaine: As a natural methyl donor and osmotic regulator, it can maintain the osmotic pressure balance of cells, protect microbial cells from environmental stress, improve the survival rate and stability of microorganisms, and also has the function of promoting protein synthesis and fat metabolism.

[0013] Peptone: It is a high-quality protein source that provides the necessary nitrogen source and amino acids for the growth and reproduction of microorganisms. It can promote the rapid proliferation of microorganisms and improve fermentation efficiency. It also has the function of protecting microbial cells.

[0014] (Trehalose-6-phosphate)-beeswax complex: As the inner coating material, trehalose-6-phosphate has good water retention and thermal stability, which can protect microbial cells from damage by heat and osmotic pressure. Beeswax has good oxygen barrier properties and can prevent oxygen from damaging microorganisms.

[0015] Glycerol citrate-gum arabic complex: As the outer coating material, glycerol citrate has good emulsifying properties, which can improve the dispersibility and stability of microcapsules, while gum arabic has good film-forming properties and pH buffering capacity.

[0016] (3) Beneficial effects Compared with the prior art, the present invention has the following beneficial effects: 1. The network structure formed by hydrogen bonding of the outer layer of glyceryl citrate-gum arabic complex and the electrostatic network constructed by the inner layer (trehalose-6-phosphate)-beeswax complex ensure that microorganisms are gradually activated under the most suitable conditions, significantly improving the survival rate and utilization efficiency of active substances.

[0017] 2. Through the hydrogen bond network between citric acid glyceride and trehalose-6-phosphate, and the coordination effect between gum arabic and trehalose-6-phosphate, an amphiphilic transition region is formed, which not only enhances the structural stability of the microcapsules, but also effectively regulates the water penetration rate, avoiding the problem of sudden structural collapse that is prone to occur in traditional microcapsules during use. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a SEM image of the fermentation preparation of Example 1 of the present invention. DETAILED DESCRIPTION

[0019] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0020] The test equipment and preparations of the examples described below are as follows: Electronic balance (Sartorius, Germany), electric blast constant temperature dryer (Shanghai Fomar Laboratory Equipment), electric constant temperature water bath (Jiangsu Kedao), freeze dryer (Shanghai Jipu), magnetic stirrer (Shanghai Meiyingpu), high-speed centrifuge (Hunan Keda), vacuum drying oven (Wuxi Marui), ultrasonic processor (Anhui Zhongke), spray dryer (Jiangsu Shuntong), fluidized bed coater (Changzhou Pinge), pulverizer (Shandong Dingkun), constant temperature incubator (Qingdao Mingcheng); chemicals and reagents were purchased from Sigma-Aldrich.

[0021] Example 1: This example discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0022] The core material layer comprises the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 28 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 21 parts of glyceryl citrate-gum arabic complex.

[0023] When fermentation preparations enter the feed production environment, Figure 1As shown in Figure 1, its structure undergoes an orderly dynamic change process, which mainly includes three stages: outer layer dissolution, transition zone regulation, and inner layer release.

[0024] The first stage is the dissolution of the outer layer. The outer layer of the glyceryl citrate-gum arabic complex first comes into contact with the environment. Due to its excellent water solubility and highly branched polysaccharide structure, gum arabic begins to dissolve slowly under suitable temperature and humidity conditions. The hydroxyl groups of glyceryl citrate and the carboxyl groups in the gum arabic molecules form a network structure through hydrogen bonding, which can ensure the penetration of water molecules and provide a certain mechanical strength to avoid sudden impact of the external environment on the inner layer.

[0025] The second stage involves the regulation of the transition zone. A unique transition zone is formed between the outer and inner layers, which is mainly due to two aspects of molecular action: on the one hand, the hydrophilic groups of citric acid glyceride form a hydrogen bond network with the phosphate groups of trehalose-6-phosphate, and its hydrophobic segments interact hydrophobically with the fatty acid esters of beeswax; on the other hand, the carboxyl groups in the gum arabic molecules form a coordination effect with the phosphate groups of trehalose-6-phosphate. This amphiphilic structure not only enhances the interlayer bonding force, but also regulates the penetration rate of water molecules to prevent the sudden collapse of the inner layer structure.

[0026] The third stage is the release of the inner protective layer. When the outer layer gradually dissolves, the inner layer (trehalose-6-phosphate)-beeswax complex begins to work. Under the heat generated by microbial fermentation, the beeswax gradually softens, but due to the phosphate group in the trehalose-6-phosphate molecule (-PO4H under pH 6.5-7.0), the beeswax gradually softens. - ) forms an electrostatic network with the hydroxyl groups on the surface of the beeswax molecules, allowing the inner layer to remain relatively stable when the temperature rises, which can both protect the active substances in the core layer and allow the gradual exchange of nutrients and metabolites.

[0027] During the entire fermentation process, as the temperature rises and water infiltrates, the mesh structure formed in the outer layer contains a large number of nano-scale channels, which is conducive to gas exchange and water regulation; the microporous structure formed by the phase change of the inner layer is regulated by the outer layer, ensuring that Lactobacillus plantarum and Candida utilis can be gradually released and grow under suitable conditions. This structural design ultimately achieves the slow release and protection of the active ingredients of the fermentation preparation during the feed production process, thereby improving the quality of feed fermentation.

[0028] The preparation method of the (trehalose-6-phosphate)-beeswax complex comprises: S11. The beeswax is heated to 65-70 ° C to obtain melted beeswax, the melted beeswax is added to a phosphate buffer at 65-70 ° C, and then stirred at 400-500 rpm for 30-40 min to obtain a beeswax emulsion; S12. adding trehalose-6-phosphate to phosphate buffer to obtain a trehalose-6-phosphate solution, adding the trehalose-6-phosphate solution dropwise to the beeswax emulsion, and reacting at 300-400 rpm for 2-3 hours to obtain a reactant; S13. Centrifuge the reactant at 3000-4000 rpm for 15-20 min to obtain a precipitate, wash the precipitate with deionized water for 3-4 times to obtain a washed product, vacuum dry the washed product at 40-50° C. for 12-24 h to obtain a dried product, grind the dried product into powder to obtain a (trehalose-6-phosphate)-beeswax complex.

[0029] Furthermore, the preparation method of the glycerol citrate-gum arabic complex comprises: S21. Adding gum arabic to deionized water and stirring at 40-50° C. for 2-3 h to obtain a gum arabic solution; S22. Adding glyceryl citrate to anhydrous ethanol to obtain glyceryl citrate solution; S23. Slowly add the citric acid glyceride solution to the gum arabic solution at a dropping speed of 2 to 3 drops / s. After the dropping is completed, stir at 300 to 400 rpm for 4 to 5 hours to obtain a reactant; S24. The reactant is ultrasonically treated for 15 to 20 minutes to obtain an ultrasonic reactant, and the ultrasonic reactant is centrifuged at 4000 to 5000 rpm for 20 to 25 minutes to obtain a centrifuged reactant; S25. Wash the centrifuged reactant 3 to 5 times with a mixed solvent consisting of water and ethanol in a volume ratio of 1:1 to obtain a washed reactant, freeze-dry the washed reactant for 36 to 48 hours to obtain a dried reactant, grind the dried reactant into powder to obtain a glyceryl citrate-gum arabic complex.

[0030] A preparation process of a fermentation preparation in a feed production process, applied to the fermentation preparation in the feed production process, comprises the following steps: S31. The Lactobacillus plantarum was activated and cultured in MRS medium at 37°C for 24 hours to obtain the cultured Lactobacillus plantarum, the cultured Lactobacillus plantarum was centrifuged at 4000-5000 rpm for 10-15 minutes to obtain Lactobacillus plantarum, and the Lactobacillus plantarum was washed 2-3 times with phosphate buffer to obtain the activated Lactobacillus plantarum; S32. The utilis strain was activated and cultured in YPD medium at 28°C for 48h to obtain cultured utilis Candida, the cultured utilis Candida was centrifuged at 3500-4000rpm for 15-20min to obtain utilis Candida cells, and the utilis Candida cells were washed 2-3 times with phosphate buffer to obtain activated utilis Candida; S33. The activated Lactobacillus plantarum and Candida utilis cells were mixed to obtain a mixed cell, the mixed cell was added to a betaine solution to obtain a first mixed solution, the first mixed solution was added with peptone to obtain a second mixed solution, the second mixed solution was stirred at 300 to 400 rpm for 30 to 40 min to obtain a stirred mixed solution, the pH of the stirred mixed solution was adjusted to 6.8 to 7.2 to obtain a core liquid; S34. The prepared core liquid was spray dried to obtain initial particles at an air inlet temperature of 45 to 50°C, an air outlet temperature of 35 to 40°C, and an atomization pressure of 0.2 MPa; S35. Add (trehalose-6-phosphate)-beeswax complex to deionized water and stir at 10000-15000 rpm for 5-10 min to obtain (trehalose-6-phosphate)-beeswax complex dispersion, place core particles in a fluidized bed, and spray the (trehalose-6-phosphate)-beeswax complex dispersion from above at a spray pressure of 0.15-0.20 MPa, an inlet air temperature of 35-40° C., a bed temperature of 30-35° C., an atomization wind speed of 35-45 m / s, and a spray rate of 8-12 mL / min to obtain an inner coating structure; S36. Add the glyceryl citrate-gum arabic complex to deionized water and ultrasonically disperse it for 15 to 20 minutes to obtain a glyceryl citrate-gum arabic complex dispersion, place the inner coating structure in a fluidized bed, and spray the glyceryl citrate-gum arabic complex dispersion from above at a spray pressure of 0.12 to 0.15 MPa, an air inlet temperature of 40 to 45° C., a bed temperature of 35 to 38° C., an atomization wind speed of 30 to 40 m / s, and a spray rate of 6 to 10 mL / min to obtain an outer coating structure; S37. The outer coating structure is vacuum dried at 35-40°C for 4-6 hours to obtain a dried product, and the dried product is sieved through an 80-mesh sieve to obtain a fermentation preparation.

[0031] Example 2: This example discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0032] The core material layer comprises the following raw materials in parts by weight: 15 parts of Lactobacillus plantarum, 20 parts of Candida utilis, 8 parts of betaine, and 10 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 25 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 15 parts of glyceryl citrate-gum arabic complex.

[0033] The preparation methods of the (trehalose-6-phosphate)-beeswax complex and the citric acid glyceride-gum arabic complex in this example are consistent with those in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0034] Example 3: This example discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0035] The core material layer comprises the following raw materials in parts by weight: 25 parts of Lactobacillus plantarum, 30 parts of Candida utilis, 12 parts of betaine, and 15 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 35 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 25 parts of glyceryl citrate-gum arabic complex.

[0036] The preparation methods of the (trehalose-6-phosphate)-beeswax complex and the citric acid glyceride-gum arabic complex in this example are consistent with those in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0037] Example 4: This example discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0038] The core material layer includes the following raw materials in parts by weight: 20 parts of Lactobacillus plantarum, 25 parts of Candida utilis, 10 parts of betaine, and 12.5 parts of peptone; the inner layer of the coating wall material layer includes the following raw materials in parts by weight: 30 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer includes the following raw materials in parts by weight: 20 parts of glyceryl citrate-gum arabic complex.

[0039] The preparation methods of the (trehalose-6-phosphate)-beeswax complex and the citric acid glyceride-gum arabic complex in this example are consistent with those in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0040] Example 5: This example discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is a citrate glyceride-gum arabic complex.

[0041] The core material layer includes the following raw materials in parts by weight: 16 parts of Lactobacillus plantarum, 28 parts of Candida utilis, 11 parts of betaine, and 13 parts of peptone; the inner layer of the coating wall material layer includes the following raw materials in parts by weight: 31 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer includes the following raw materials in parts by weight: 24 parts of glyceryl citrate-gum arabic complex.

[0042] The preparation methods of the (trehalose-6-phosphate)-beeswax complex and the citric acid glyceride-gum arabic complex in this example are consistent with those in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0043] Control group 1: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is beeswax, and the outer layer is a citric acid glyceride-gum arabic complex.

[0044] The core material layer comprises the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 28 parts of beeswax; and the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 21 parts of glyceryl citrate-gum arabic complex.

[0045] The preparation method of the beeswax and glyceryl citrate-gum arabic complex of this embodiment is consistent with that of embodiment 1. The preparation process of the fermentation preparation in a feed production process of this embodiment is consistent with that of embodiment 1.

[0046] Control group 2: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is trehalose-6-phosphate, and the outer layer is a citrate glyceride-gum arabic complex.

[0047] The core material layer comprises the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 28 parts of trehalose-6-phosphate; and the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 21 parts of citric acid glyceride-gum arabic complex.

[0048] The preparation method of the trehalose-6-phosphate and citric acid glyceride-gum arabic complex of this embodiment is consistent with that of embodiment 1. The preparation process of the fermentation preparation in a feed production process of this embodiment is consistent with that of embodiment 1.

[0049] Control group 3: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone, and the coating wall material layer is a glyceryl citrate-gum arabic complex.

[0050] The core material layer comprises the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the coating wall material layer comprises the following raw materials in parts by weight: 21 parts of citric acid glyceride-arabic gum complex.

[0051] The preparation method of the glycerol citrate-gum arabic complex of this embodiment is consistent with that of embodiment 1. The preparation process of the fermentation preparation in a feed production process of this embodiment is consistent with that of embodiment 1.

[0052] Control group 4: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is gum arabic.

[0053] The core material layer includes the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the inner layer of the coating wall material layer includes the following raw materials in parts by weight: 28 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer includes the following raw materials in parts by weight: 21 parts of gum arabic.

[0054] The preparation method of the (trehalose-6-phosphate)-beeswax complex and gum arabic in this example is consistent with that in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0055] Control group 5: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone. The coating wall material layer includes an inner layer and an outer layer. The inner layer is a (trehalose-6-phosphate)-beeswax complex, and the outer layer is glycerol citrate.

[0056] The core material layer includes the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the inner layer of the coating wall material layer includes the following raw materials in parts by weight: 28 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer includes the following raw materials in parts by weight: 21 parts of citric acid glyceride.

[0057] The preparation method of the (trehalose-6-phosphate)-beeswax complex and citric acid glyceride in this example is consistent with that in Example 1. The preparation process of the fermentation preparation in a feed production process in this example is consistent with that in Example 1.

[0058] Control group 6: This embodiment discloses a fermentation preparation for a feed production process, which is composed of microcapsules. The microcapsules include a core material layer and a coating wall material layer. The core material layer is Lactobacillus plantarum, Candida utilis, betaine and peptone, and the coating wall material layer is a (trehalose-6-phosphate)-beeswax complex.

[0059] The core material layer comprises the following raw materials in parts by weight: 18 parts of Lactobacillus plantarum, 22 parts of Candida utilis, 10 parts of betaine, and 12 parts of peptone; the coating wall material layer comprises the following raw materials in parts by weight: 28 parts of (trehalose-6-phosphate)-beeswax complex.

[0060] The preparation method of the (trehalose-6-phosphate)-beeswax complex of this embodiment is consistent with that of embodiment 1. The preparation process of the fermentation preparation in a feed production process of this embodiment is consistent with that of embodiment 1.

[0061] Test verification: 1. Outer layer dissolution behavior experiment: 2.0 g of the fermentation preparation of the example group and the control group were weighed and placed in a 250 mL conical flask, 100 mL of phosphate buffer with a pH of 6.8 preheated to 25°C was added, and the mixture was shaken at 150 rpm. 5 mL of the mixture was sampled at designated time points (0, 15, 30, 60, 120, and 240 minutes), filtered through a 0.45 μm filter membrane, and stored in 2 mL centrifuge tubes to determine the citric acid glyceride content and the gum arabic content.

[0062] The buffer was a phosphate buffer with a pH of 6.8, the temperature gradient was set at 25°C, 35°C, and 45°C, the sampling time points were 0, 15, 30, 60, 120, and 240 minutes, and the citrate glyceride content and gum arabic content were determined.

[0063] 2. Inner layer stability: Place the sample in a programmed temperature-controlled box with the temperature program set as follows: maintain at 25°C for 2 hours, increase to 45°C at 2°C / min, maintain at 45°C for 2 hours, decrease to 25°C at 2°C / min, then weigh 10 mg of the sample into an aluminum pan, heat up at a rate of 10°C / min, scan in the range of 20-80°C, and record the thermal enthalpy change.

[0064] 3. Moisture penetration test: Fix the sample on the upper end of the moisture permeable cup, place a desiccant on the lower end, put the device into the environment, weigh it every 4 hours, and calculate the water vapor transmission rate.

[0065] 4. Storage stability: The storage conditions are 25°C and relative humidity 60%. The colony counts are measured at 0, 7, 14, 30, 60, and 90 days. Colony counts: 0.5 g of the sample is dissolved in sterile water, graded dilutions are performed, and the samples are spread on the plates for counting.

[0066] 5. Release under simulated fermentation conditions: temperature 37°C, relative humidity 75%, pH 6.5, sampling time 0, 2, 4, 6, 8, 12, 24 hours, and determination of viable counts.

[0067]

[0068] The citric acid glyceride release rate in Table 1 and the gum arabic release rate in Table 2 show that Examples 1 to 5 exhibit a relatively ideal sustained-release effect, with a smooth release curve and a release rate of 82 to 89% at 240 min. In contrast, Control Group 3 showed an excessively fast release rate (exceeding 95% at 240 min), and Control Group 5 also showed a relatively fast release (about 95% at 240 min), which indicates that the composite coating material in the embodiment can better control the release rate. Table 3 shows the DSC analysis results showing that the enthalpy change of Examples 1 to 5 decreased slightly during the cycle (about 5 to 10%), and the enthalpy change of Control Groups 1 to 2 decreased significantly (about 35 to 40%), which indicates that the embodiment formula has better thermal stability and structural integrity. Table 4 shows the water vapor permeability. The water vapor permeability of Examples 1 to 5 is significantly lower than that of the control group. At 24 h, the permeability of the embodiment group is between 13 and 18 g / m 2 ·24h, the permeability of the control group was generally higher than 20 g / m 2 ·24h, indicating that the example formula has better moisture barrier performance. Table 5 is a study of storage stability. The live bacteria count of the example group is still maintained at 7.9~8.5 lg CFU / g after 90 days, and the live bacteria count of the control group dropped to 5.5~7.0 lgCFU / g after 90 days, indicating that the example formula can better protect the probiotics. Table 6 is the release of live bacteria count under simulated fermentation conditions. The example group showed an ideal sustained release characteristic, reaching a peak at 24h, and the control group reached a peak at 8~12h, and then began to decline, indicating that the example formula can better control the release of strains and is conducive to continuous fermentation. Comprehensive analysis shows that the formulas of Examples 1 to 5 achieve better sustained release effects, higher thermal stability, lower water permeability, and better strain protection capabilities through composite coating materials (inner layer (trehalose-6-phosphate)-beeswax, outer layer citric acid glyceride-arabic gum), among which Example 1 has the best comprehensive performance.

[0069] Finally, it should be noted that: Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A fermentation preparation in a feed production process, characterized in that: The microcapsule comprises a core material layer and a coating wall material layer. The core material layer comprises plant lactobacillus, Candida utilis, betaine and peptone. The coating wall material layer comprises an inner layer and an outer layer. The inner layer comprises a (trehalose-6-phosphate)-beeswax complex, and the outer layer comprises a citric acid glyceride-arabic gum complex.

2. A fermentation preparation for feed production process according to claim 1, characterized in that: The core material layer comprises the following raw materials in parts by weight: 15-25 parts of Lactobacillus plantarum, 20-30 parts of Candida utilis, 8-12 parts of betaine, and 10-15 parts of peptone; the inner layer of the coating wall material layer comprises the following raw materials in parts by weight: 25-35 parts of (trehalose-6-phosphate)-beeswax complex; the outer layer of the coating wall material layer comprises the following raw materials in parts by weight: 15-25 parts of glyceryl citrate-gum arabic complex.

3. A fermentation preparation for feed production process according to claim 2, characterized in that: The preparation method of the (trehalose-6-phosphate)-beeswax complex comprises: S11. The beeswax is heated to 65-70 ° C to obtain melted beeswax, the melted beeswax is added to a phosphate buffer at 65-70 ° C, and then stirred at 400-500 rpm for 30-40 min to obtain a beeswax emulsion; S12. adding trehalose-6-phosphate to phosphate buffer to obtain a trehalose-6-phosphate solution, adding the trehalose-6-phosphate solution dropwise to the beeswax emulsion, and reacting at 300-400 rpm for 2-3 hours to obtain a reactant; S13. Centrifuge the reactant at 3000-4000 rpm for 15-20 min to obtain a precipitate, wash the precipitate with deionized water for 3-4 times to obtain a washed product, vacuum dry the washed product at 40-50° C. for 12-24 h to obtain a dried product, grind the dried product into powder to obtain a (trehalose-6-phosphate)-beeswax complex.

4. A fermentation preparation for feed production process according to claim 2, characterized in that: The preparation method of the glyceryl citrate-arabic gum complex comprises: S21. Adding gum arabic to deionized water and stirring at 40-50° C. for 2-3 h to obtain a gum arabic solution; S22. Adding glyceryl citrate to anhydrous ethanol to obtain glyceryl citrate solution; S23. Slowly add the glycerol citrate solution to the gum arabic solution at a dropping speed of 2 to 3 drops / s. After the dropping is completed, stir at 300 to 400 rpm for 4 to 5 hours to obtain a reactant; S24. The reactant is ultrasonically treated for 15 to 20 minutes to obtain an ultrasonic reactant, and the ultrasonic reactant is centrifuged at 4000 to 5000 rpm for 20 to 25 minutes to obtain a centrifuged reactant; S25. Wash the centrifuged reactant 3 to 5 times with a mixed solvent consisting of water and ethanol in a volume ratio of 1:1 to obtain a washed reactant, freeze-dry the washed reactant for 36 to 48 hours to obtain a dried reactant, grind the dried reactant into powder to obtain a glyceryl citrate-gum arabic complex.

5. A process for preparing a fermentation preparation in a feed production process, which is applied to prepare a fermentation preparation in a feed production process as claimed in any one of claims 1 to 4, characterized in that: The process comprises the following steps: S31. The Lactobacillus plantarum was activated and cultured in MRS medium at 37°C for 24 hours to obtain the cultured Lactobacillus plantarum, the cultured Lactobacillus plantarum was centrifuged at 4000-5000 rpm for 10-15 minutes to obtain Lactobacillus plantarum, and the Lactobacillus plantarum was washed 2-3 times with phosphate buffer to obtain the activated Lactobacillus plantarum; S32. The utilis strain was activated and cultured in YPD medium at 28°C for 48h to obtain cultured utilis Candida, the cultured utilis Candida was centrifuged at 3500-4000rpm for 15-20min to obtain utilis Candida cells, and the utilis Candida cells were washed 2-3 times with phosphate buffer to obtain activated utilis Candida; S33. The activated Lactobacillus plantarum and Candida utilis cells were mixed to obtain a mixed cell, the mixed cell was added to a betaine solution to obtain a first mixed solution, the first mixed solution was added with peptone to obtain a second mixed solution, the second mixed solution was stirred at 300 to 400 rpm for 30 to 40 min to obtain a stirred mixed solution, the pH of the stirred mixed solution was adjusted to 6.8 to 7.2 to obtain a core liquid; S34. The prepared core liquid was spray dried to obtain initial particles at an air inlet temperature of 45 to 50°C, an air outlet temperature of 35 to 40°C, and an atomization pressure of 0.2 MPa; S35. Add (trehalose-6-phosphate)-beeswax complex to deionized water and stir at 10000-15000 rpm for 5-10 min to obtain (trehalose-6-phosphate)-beeswax complex dispersion, place core particles in a fluidized bed, and spray the (trehalose-6-phosphate)-beeswax complex dispersion from above at a spray pressure of 0.15-0.20 MPa, an inlet air temperature of 35-40° C., a bed temperature of 30-35° C., an atomization wind speed of 35-45 m / s, and a spray rate of 8-12 mL / min to obtain an inner coating structure; S36. Add the glyceryl citrate-gum arabic complex to deionized water and ultrasonically disperse it for 15 to 20 minutes to obtain a glyceryl citrate-gum arabic complex dispersion, place the inner coating structure in a fluidized bed, and spray the glyceryl citrate-gum arabic complex dispersion from above at a spray pressure of 0.12 to 0.15 MPa, an air inlet temperature of 40 to 45° C., a bed temperature of 35 to 38° C., an atomization wind speed of 30 to 40 m / s, and a spray rate of 6 to 10 mL / min to obtain an outer coating structure; S37. The outer coating structure is vacuum dried at 35-40°C for 4-6 hours to obtain a dried product, and the dried product is sieved through an 80-mesh sieve to obtain a fermentation preparation.

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