CN117581941A - Stable lactobacillus preparation and preparation method and application thereof - Google Patents

Stable lactobacillus preparation and preparation method and application thereof Download PDF

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Publication number
CN117581941A
CN117581941A CN202311454801.0A CN202311454801A CN117581941A CN 117581941 A CN117581941 A CN 117581941A CN 202311454801 A CN202311454801 A CN 202311454801A CN 117581941 A CN117581941 A CN 117581941A
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Prior art keywords
lactobacillus
lactic acid
acid bacteria
carrier
stable
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Inventor
王绍云
陈祺琛
陈新伟
张茜茜
王启军
辜玲芳
邱权
杨雄振
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Sunhy Technology Hubei Co ltd
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Sunhy Technology Hubei Co ltd
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Priority to CN202311454801.0A priority Critical patent/CN117581941A/en
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/10—Shaping or working-up of animal feeding-stuffs by agglomeration; by granulation, e.g. making powders
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
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    • A23L3/349—Organic compounds containing oxygen with singly-bound oxygen
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
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    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3481—Organic compounds containing oxygen
    • A23L3/3508—Organic compounds containing oxygen containing carboxyl groups
    • A23L3/3517—Carboxylic acid esters
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3526—Organic compounds containing nitrogen
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3562—Sugars; Derivatives thereof
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/23—Lactobacillus acidophilus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Abstract

The invention belongs to the technical field of probiotic preparations, and particularly provides a preparation method of a stable lactobacillus preparation, which comprises the following steps: (1) centrifuging lactobacillus fermentation liquor to obtain concentrated bacteria liquor; (2) Fully mixing the concentrated bacterial liquid with a carrier, and standing to obtain a lactic acid bacteria matrix; (3) Adding a protective agent into the lactobacillus matrix, and uniformly mixing; (4) And (3) extruding and granulating the mixed material obtained in the step (3), and drying, and forming a coating layer on the surface of the granules by using a coating material to obtain the stable lactobacillus preparation. According to the method, concentrated bacterial liquid and the carrier are fully mixed and then subjected to stabilization treatment at a proper temperature, so that the bacterial body is gradually adapted to the carrier and the environmental conditions, the aim of improving the tolerance and stress resistance of the bacterial body is fulfilled, the survival of the bacterial body in the subsequent treatment process is facilitated, meanwhile, a proper protective agent is added, the storage stability of the product can be improved, the shelf life of the product is prolonged, and the survival rate of the product in the granulating process of the pellet feed can be improved.

Description

Stable lactobacillus preparation and preparation method and application thereof
Technical Field
The invention belongs to the technical field of probiotic preparations, and particularly relates to a stable lactobacillus preparation and a preparation method and application thereof.
Background
Lactic acid bacteria (lactic acid bacteria, LAB) are a generic term for a class of bacteria that can utilize fermentable carbohydrates to produce large amounts of lactic acid. The bacteria are widely distributed in nature, most of the bacteria are the bacterial groups which are indispensable in human bodies and have important physiological functions except very few bacteria, and the bacteria are widely existing in intestinal tracts of the human bodies. Lactic acid bacteria have extremely high application value in important fields closely related to human life, such as industry, agriculture and animal husbandry, food, medicine and the like.
A large number of normal microbial flora exists in the gastrointestinal tract of animals, and the microbial flora and a host establish a mutually dependent and mutually restricted microecological relationship to form a uniform whole body so as to form a dynamic balance system. Under normal physiological conditions of animal organisms, the gastrointestinal microbial flora plays an important role in maintaining animal health, nutrient utilization, colonization resistance and the like. Lactic acid bacteria are the usual bacteria for the intestinal tract. After the livestock and poultry take the lactobacillus, the environment in intestinal tracts can be changed, the reproduction of harmful bacteria can be inhibited, the balance of gastrointestinal flora can be adjusted, the disease resistance of the livestock and poultry can be enhanced, and the survival rate and daily gain can be improved. At present, lactobacillus preparations are widely applied to various fields of pig, chicken, cattle, sheep, aquaculture and the like.
The traditional production method of the lactobacillus agent is to carry out high-density fermentation on the lactobacillus, concentrate the lactobacillus by a centrifuge, add a certain amount of protective agent, mix the lactobacillus with the protective agent, and freeze-dry the lactobacillus to obtain the fungus powder. The lactobacillus preparation prepared by the method is generally poor in stability, easy to inactivate in the normal-temperature storage and transportation process and high in cost, so that the lactobacillus preparation is difficult to be widely popularized and used in livestock breeding. Therefore, a stable and low-cost lactobacillus product is developed, and the method has important significance for safety, high efficiency and environmental protection of livestock production.
Disclosure of Invention
The invention aims to solve the problems that the stability of a lactobacillus preparation in the prior art is poor and high-efficiency utilization is difficult to realize.
Therefore, the invention provides a preparation method of a stable lactobacillus preparation, which comprises the following steps:
(1) Centrifuging the lactobacillus fermentation liquor to obtain concentrated bacterial liquor;
(2) Fully mixing the concentrated bacterial liquid with a carrier, and standing to obtain a lactic acid bacteria matrix;
(3) Adding a protective agent into the lactobacillus matrix, and uniformly mixing;
(4) And (3) extruding and granulating the mixed material obtained in the step (3), and drying, and forming a coating layer on the surface of the granules by using a coating material to obtain the stable lactobacillus preparation.
Specifically, the carrier in the step (2) includes a mass ratio of (30-20): (20-10): (10-5): bran, corn husk, starch and guar gum of (5-1).
Specifically, the mass ratio of the concentrated bacterial liquid to the carrier in the step (2) is 1: (0.5-2).
Specifically, the concentrated bacterial liquid in the step (2) is fully mixed with the carrier and then is stabilized for 24-48 hours at the temperature of 30-35 ℃.
Specifically, the protective agent in the step (3) comprises 0.5-1% of trehalose, 1-2% of xylooligosaccharide, 1-2% of sodium glutamate, 1-2% of inositol and 0.5-1% of hyaluronic acid by mass of the lactobacillus matrix.
Specifically, in the step (4), a high-pressure screw extruder is adopted for extrusion granulation, and the length-diameter ratio of the screw is (4-8): 1.
Specifically, the coating material in the step (4) comprises one or more of palm stearin, stearic acid, hydrogenated oil, mono/diglyceride and beeswax.
Specifically, after the coating materials are melted in the step (4), a coating machine is adopted to spray the coating materials on the surfaces of the particles to form a coating layer, the coating weight is increased by 10-40%, and the material temperature in the coating process is controlled at 40-50 ℃.
Compared with the prior art, the invention has the following advantages and beneficial effects:
according to the preparation method of the stable lactobacillus preparation, the concentrated bacterial liquid and the carrier are fully mixed and then subjected to stabilization treatment at a proper temperature, so that the bacterial body is gradually adapted to the used carrier and environmental conditions, the purposes of improving the tolerance and stress resistance of the bacterial body are achieved, the survival of the bacterial body in the subsequent treatment process is facilitated, meanwhile, a proper protective agent is added, the storage stability of the product can be improved, the shelf life of the product is prolonged, and the survival rate of the product in the granulating process of the pellet feed can be improved. In addition, the high-pressure screw extruder is adopted during granulation instead of a common granulator, so that the lactobacillus particles with high compactness and high compactness are obtained, and the situation that steam enters the particles in the feed granulating process can be prevented is ensured, so that the effect of protecting internal thalli is achieved, and further, the product has higher viable bacteria retention rate in the feed granulating process is ensured. The ester coating material is adopted for coating, so that oxygen and moisture in the air can be further isolated, the activity of thalli is better ensured, and the shelf life of products is prolonged; in the feed granulating process, the coating layer formed by the ester coating material can also prevent or delay steam from entering the inside of the granule, thereby playing a role in protecting the activity of the product.
Detailed Description
The technical solutions of the present invention will be clearly and completely described in the following examples, and it is obvious that the described examples are only some examples of the present invention, but not all examples. Although representative embodiments of the present invention have been described in detail, those skilled in the art to which the invention pertains will appreciate that various modifications and changes can be made without departing from the scope of the invention. Accordingly, the scope of the invention should not be limited to the embodiments, but should be defined by the appended claims and equivalents thereof.
The invention provides a preparation method of a stable lactobacillus preparation, which comprises the following steps:
(1) Centrifuging the lactobacillus fermentation liquor to obtain concentrated bacterial liquor; the solid content of the concentrated bacterial liquid is preferably 20-30%.
(2) Bran, corn husk, starch and guar gum are mixed according to the mass ratio of (30-20): (20-10): (10-5): and (5-1) fully mixing to obtain the carrier, wherein the mesh number of all raw materials is smaller than 80 mesh.
Mixing concentrated bacterial liquid with a carrier according to the following ratio of 1: (0.5-2), placing in a 30-35 ℃ incubator, and standing for stabilization treatment for 24-48h to obtain lactobacillus matrix.
The guar gum in the carrier is a colloid substance, so that the prepared lactobacillus particles still have high compactness and high compactness after being dried, and the obtained lactobacillus products can be prevented from entering the particles by steam in the feed granulating process, so that the effect of protecting internal thalli is achieved, and the products are further ensured to have higher viable bacteria retention rate in the feed granulating process.
(3) Adding 0.5-1% of trehalose, 1-2% of xylooligosaccharide, 1-2% of sodium glutamate, 1-2% of inositol and 0.5-1% of hyaluronic acid into the lactobacillus matrix by mass of the lactobacillus matrix as a protective agent, uniformly mixing, adding a proper amount of carrier in the step (2) according to the water content of the carrier, and uniformly mixing to ensure that the final water content of the mixed material is 15-25%.
(4) And (3) putting the mixed material obtained in the step (3) into a high-pressure screw extruder for extrusion granulation, wherein the length-diameter ratio of the screw of the high-pressure screw extruder is (4-8): 1, preferably more than 5:1, so that higher extrusion pressure is obtained, and the compactness and compactness of the particles are ensured. The high-pressure screw extruder is preferably provided with a circulating water cooling system, and cooling water with the temperature below 5 ℃ is used for cooling during granulation. The pore diameter of the extrusion screen plate is 0.5-1mm, and the wet granules are dried in a fluidized bed at 40-45 ℃ until the water content is 2-5%.
And after the coating material is melted, spraying the coating material on the surface of the particles by a coating machine to form a coating layer, thus obtaining the stable lactobacillus preparation. The weight gain of the coating is 10-40%, and the material temperature in the coating process is controlled at 40-50 ℃.
The coating material comprises one or more of palm stearin, stearic acid, hydrogenated oil, mono/diglyceride, beeswax and other hot melt coating materials. Such coatings are typically used as slow release or overgastric etc. and no coatings for microbial products are found. The invention is used for coating lactobacillus preparation, and further isolates oxygen and moisture in the air, thereby better ensuring the activity of thalli and prolonging the shelf life of products. Meanwhile, in the feed granulating process, the coating layer formed by the ester coating material can also prevent or delay steam from entering the inside of the granule, so that the effect of protecting the activity of the product is achieved.
The effect of the method for producing the stabilized lactic acid bacteria preparation of the present invention is examined by the following specific examples.
Example 1:
the present example provides a stable lactic acid bacteria preparation prepared by the steps of:
(1) And (3) centrifuging the lactobacillus plantarum fermentation liquid obtained by high-density fermentation by using a disc type centrifuge to obtain concentrated bacterial liquid with 20% of solid content.
(2) Bran, corn husk, starch and guar gum are mixed according to the mass ratio of 20:20:10:1, fully mixing to obtain a carrier, wherein the mesh number of all raw materials is less than 80 meshes;
mixing concentrated bacterial liquid with a carrier according to the following ratio of 1:2, fully mixing the materials in a mixer, and stabilizing the materials at 30 ℃ for 48 hours to obtain the lactobacillus matrix.
(3) Adding 0.5% of trehalose, 1% of xylooligosaccharide, 2% of sodium glutamate, 1% of inositol and 0.5% of hyaluronic acid into the lactobacillus matrix by mass of the lactobacillus matrix as a protective agent, uniformly mixing, adding a carrier into the lactobacillus matrix according to the water content of the lactobacillus matrix, and uniformly mixing again to ensure that the water content of the final mixed material is 15%.
(4) And (3) putting the mixed material obtained in the step (3) into a high-pressure screw extruder for extrusion granulation, wherein the length-diameter ratio of a screw of the high-pressure screw extruder is 4:1, the aperture of an extrusion sieve plate is 0.5mm, and drying the prepared wet particles in a fluidized bed at 40 ℃ until the water content is 5%.
And melting palm stearin accounting for 10% of the weight of the dried granules, and spraying the melted palm stearin on the surfaces of the granules by using a coating machine to form a coating layer, wherein the temperature of the materials in the coating process is controlled to be 40 ℃, so that the stable lactobacillus preparation is obtained.
Comparative example 1:
taking the concentrated bacterial liquid obtained in the example 1, adding twice the mass of bran as a carrier, uniformly stirring, granulating by using a low-pressure screw extruder, and then drying in a boiling drying bed to prepare the granular lactobacillus preparation.
Example 2:
the present example provides a stable lactic acid bacteria preparation prepared by the steps of:
(1) And (3) centrifuging the lactobacillus acidophilus fermentation liquid subjected to high-density fermentation by using a disc type centrifuge to obtain concentrated bacterial liquid with 25% of solid content.
(2) Bran, corn husk, starch and guar gum are mixed according to the mass ratio of 25:15:8:2, fully mixing to obtain a carrier, wherein the mesh number of all raw materials is less than 80 meshes;
mixing concentrated bacterial liquid with a carrier according to the following ratio of 1:1, and then fully mixing the materials in a mixer, and keeping the mixture at the temperature of 32 ℃ for 36 hours to obtain the lactobacillus matrix.
(3) Adding 1% of trehalose, 1% of xylooligosaccharide, 1% of sodium glutamate, 2% of myo-inositol and 1% of hyaluronic acid into the lactobacillus matrix by mass of the lactobacillus matrix as a protective agent, uniformly mixing, adding a carrier into the lactobacillus matrix according to the water content of the lactobacillus matrix, and uniformly mixing again to ensure that the water content of the final mixed material is 20%.
(4) And (3) putting the mixed material obtained in the step (3) into a high-pressure screw extruder for extrusion granulation, wherein the length-diameter ratio of the screw of the high-pressure screw extruder is 5:1, the aperture of an extrusion sieve plate is 0.75mm, and drying the prepared wet particles in a fluidized bed at 42 ℃ until the water content is 4%.
And melting stearic acid accounting for 20% of the weight of the dried granules, and spraying the melted stearic acid on the surfaces of the dried granules by using a coating machine to form a coating layer, wherein the temperature of the material in the coating process is controlled to be 45 ℃, so that the stable lactobacillus preparation is obtained.
Comparative example 2:
taking the concentrated bacterial liquid obtained in the example 2, adding twice the mass of bran as a carrier, uniformly stirring, granulating by using a low-pressure screw extruder, and then drying in a boiling drying bed to prepare the granular lactobacillus preparation.
Example 3:
the present example provides a stable lactic acid bacteria preparation prepared by the steps of:
(1) And (3) centrifuging the high-density fermentation enterococcus faecium fermentation liquid by using a disc centrifuge to obtain concentrated bacterial liquid with the solid content of 30%.
(2) Bran, corn husk, starch and guar gum are mixed according to a mass ratio of 30:10:5:2, fully mixing to obtain the carrier, wherein the mesh number of all raw materials is less than 80 mesh.
Mixing concentrated bacterial liquid with a carrier according to the following ratio of 1: the mixture was thoroughly mixed in a mixer at a mass ratio of 0.5, and the mixture was kept at 35℃for 24 hours to obtain a stabilized lactic acid bacterium substrate.
(3) Adding 0.5% of trehalose, 2% of xylooligosaccharide, 2% of sodium glutamate, 1% of inositol and 0.5% of hyaluronic acid into the lactobacillus matrix by mass of the lactobacillus matrix as a protective agent, uniformly mixing, adding a carrier into the lactobacillus matrix according to the water content of the lactobacillus matrix, and uniformly mixing again to ensure that the water content of the final mixed material is 25%.
(4) And (3) putting the mixed material obtained in the step (3) into a high-pressure screw extruder for extrusion granulation, wherein the length-diameter ratio of a screw of the high-pressure screw extruder is 8:1, the aperture of an extrusion sieve plate is 1mm, and drying the prepared wet particles in a fluidized bed at 45 ℃ until the water content is 2%.
And melting the mono/diglyceride accounting for 40% of the weight of the dried granules, and spraying the melted mono/diglyceride on the surfaces of the granules by using a coating machine to form a coating layer, wherein the temperature of the materials in the coating process is controlled to be 50 ℃, so as to obtain the stable lactobacillus preparation.
Comparative example 3:
taking the concentrated bacterial liquid obtained in the example 3, adding twice the mass of bran as a carrier, uniformly stirring, granulating by using a low-pressure screw extruder, and then drying in a boiling drying bed to prepare the granular lactobacillus preparation.
Example 4:
the present example provides a stable lactic acid bacteria preparation prepared by the steps of:
(1) And (3) centrifuging the high-density fermented pediococcus pentosaceus fermentation liquor by a disc type centrifuge to obtain concentrated bacterial liquor with 25% of solid content.
(2) Bran, corn husk, starch and guar gum are mixed according to the mass ratio of 20:10:7:3, fully mixing to obtain the carrier, wherein the mesh number of all raw materials is less than 80 mesh.
Mixing concentrated bacterial liquid with a carrier according to the proportion of 1:1, and then, the mixture is fully mixed in a mixer and is kept at 33 ℃ for 40 hours to obtain the stabilized lactobacillus matrix.
(3) Adding 1% of trehalose, 1% of xylooligosaccharide, 1% of sodium glutamate, 2% of myo-inositol and 0.5% of hyaluronic acid into the lactobacillus matrix by mass of the lactobacillus matrix as a protective agent, uniformly mixing, adding a carrier into the lactobacillus matrix according to the water content of the lactobacillus matrix, and uniformly mixing to ensure that the water content of the final mixed material is 20%.
(4) And (3) putting the mixed material obtained in the step (3) into a high-pressure screw extruder for extrusion granulation, wherein the length-diameter ratio of a screw of the high-pressure screw extruder is 6:1, the aperture of an extrusion sieve plate is 1mm, and drying the prepared wet particles in a fluidized bed at 40 ℃ until the water content is 5%.
Melting Cera flava of 20% of the weight of the dried granule, spraying on the granule surface with a coating machine to form a coating layer, and controlling the material temperature at 40deg.C during coating process to obtain stable lactobacillus preparation.
Comparative example 4:
taking the concentrated bacterial liquid obtained in the example 4, adding twice the mass of bran as a carrier, uniformly stirring, granulating by using a low-pressure screw extruder, and then drying in a boiling drying bed to prepare the granular lactobacillus preparation.
Comparative example 5:
the lactobacillus preparation is prepared by adopting the same method as in example 1, wherein the step (2) is to mix bran, corn husk, starch and guar gum according to a mass ratio of 20:20:10:1, fully mixing to obtain a carrier, wherein the mesh number of all raw materials is less than 80 meshes;
mixing concentrated bacterial liquid with a carrier according to the following ratio of 1:2, fully mixing the materials in a mixer without stabilizing treatment process, and directly carrying out the step (3) on the obtained lactobacillus matrix.
Comparative example 6:
the same method as in example 1 was used to prepare a lactic acid bacteria preparation, except that in step (4), the mixed material obtained in step (3) was fed into a high-pressure screw extruder for extrusion granulation, the aspect ratio of the screw of the high-pressure screw extruder was 5:1, the aperture of the extrusion screen plate was 0.5mm, and the wet granules thus prepared were placed in a fluidized bed and dried at 40 ℃ until the moisture was 5%, to obtain a lactic acid bacteria preparation.
Comparative example 7:
a lactic acid bacteria preparation was prepared in the same manner as in example 1, except that the carrier comprises a mass ratio of 20:20:10, wheat bran, corn husk and starch.
Example 5:
the lactic acid bacteria preparations prepared in examples 1 to 4 and comparative examples 1 to 7 were evaluated for stability and heat resistance.
The stability evaluation method of the lactic acid bacteria preparation is as follows:
packaging 20g lactobacillus preparation product with PE inner membrane bag, placing the packaged sample in an incubator with temperature of 37deg.C and humidity of 75%, taking out after 30 days, detecting viable count of lactobacillus, calculating viable bacteria retention rate, and evaluating storage stability according to the viable bacteria retention rate.
The initial viable count of the viable cell preparations of examples 1 to 4 and comparative examples 1 to 7 and the viable cell count after 30 days of storage at 37℃were measured, respectively, and the corresponding viable cell retention ratios were calculated, and the results are shown in Table 1.
The heat resistance evaluation method of the lactobacillus preparation in the feed pelleting process is as follows:
the lactobacillus preparation product is prepared according to the following weight percentage of 1:1000 mass ratio is added into the compound feed, and then feed pelletization is carried out, wherein the feed conditioning conditions are as follows: moisture 16.5%, temperature 80 ℃ and time 45s. The prepared feed sample is used for detecting the viable count of the lactobacillus, calculating the viable bacteria retention rate, and evaluating the heat resistance of the lactobacillus during the feed pelleting process according to the viable bacteria retention rate.
The initial viable count and the viable count after granulation of the viable cell preparations of examples 1 to 4 and comparative examples 1 to 7 were measured, respectively, and the respective viable cell retention ratios were calculated, and the results are shown in Table 1.
TABLE 1 measurement results of lactic acid bacteria preparation
As can be seen from Table 1, the preparation method of the stable lactobacillus preparation provided by the invention comprises the steps of fully mixing concentrated bacteria liquid with a carrier, and then stabilizing the mixture at a proper temperature, so that the bacteria are gradually adapted to the used carrier and environmental conditions, the purposes of improving the tolerance and stress resistance of the bacteria are achieved, the survival of the bacteria in the subsequent treatment process is facilitated, meanwhile, a proper protective agent is added, the storage stability of the product can be improved, the shelf life of the product is prolonged, and the survival rate of the product in the granulating process of the pellet feed can be improved. In addition, the high-pressure screw extruder is adopted during granulation instead of a common granulator, so that the lactobacillus particles with high compactness and high compactness are obtained, and the situation that steam enters the particles in the feed granulating process can be prevented is ensured, so that the effect of protecting internal thalli is achieved, and further, the product has higher viable bacteria retention rate in the feed granulating process is ensured. The ester coating material is adopted for coating, so that oxygen and moisture in the air can be further isolated, the activity of thalli is better ensured, and the shelf life of products is prolonged; in the feed granulating process, the coating layer formed by the ester coating material can also prevent or delay steam from entering the inside of the granule, thereby playing a role in protecting the activity of the product.
The foregoing examples are merely illustrative of the present invention and are not intended to limit the scope of the present invention, and all designs that are the same or similar to the present invention are within the scope of the present invention.

Claims (10)

1. A method for preparing a stable lactobacillus preparation, which is characterized by comprising the following steps:
(1) Centrifuging the lactobacillus fermentation liquor to obtain concentrated bacterial liquor;
(2) Fully mixing the concentrated bacterial liquid with a carrier, and standing to obtain a lactic acid bacteria matrix;
(3) Adding a protective agent into the lactobacillus matrix, and uniformly mixing;
(4) And (3) extruding and granulating the mixed material obtained in the step (3), and drying, and forming a coating layer on the surface of the granules by using a coating material to obtain the stable lactobacillus preparation.
2. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: the carrier in the step (2) comprises the following components in percentage by mass (30-20): (20-10): (10-5): bran, corn husk, starch and guar gum of (5-1).
3. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: in the step (2), the mass ratio of the concentrated bacterial liquid to the carrier is 1: (0.5-2).
4. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: and (3) fully mixing the concentrated bacterial liquid and the carrier in the step (2), and stabilizing at 30-35 ℃ for 24-48 hours.
5. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: in the step (3), the protective agent comprises 0.5-1% of trehalose, 1-2% of xylooligosaccharide, 1-2% of sodium glutamate, 1-2% of inositol and 0.5-1% of hyaluronic acid by mass of lactobacillus matrix.
6. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: and (3) extruding and granulating in the step (4) by adopting a high-pressure screw extruder, wherein the length-diameter ratio of the screw is (4-8): 1.
7. The method for preparing a stable lactic acid bacteria preparation according to claim 1, characterized in that: the wrapping material in the step (4) comprises one or more of palm stearin, stearic acid, hydrogenated oil, mono/diglyceride and beeswax.
8. The method for preparing a stable lactic acid bacteria preparation according to claim 7, characterized in that: and (3) after the coating materials are melted in the step (4), spraying the coating materials on the surfaces of the particles by a coating machine to form a coating layer, wherein the coating weight is increased by 10-40%, and the material temperature in the coating process is controlled at 40-50 ℃.
9. A stabilized lactic acid bacteria preparation prepared by the method of any one of claims 1 to 8.
10. Use of a stabilized lactic acid bacteria preparation according to claim 9 in animal feed.
CN202311454801.0A 2023-11-03 2023-11-03 Stable lactobacillus preparation and preparation method and application thereof Pending CN117581941A (en)

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