Bifidobacterium metagen as well as preparation method and application thereof
By mixing Bifidobacterium cell contents with enzymatic fiber materials, Bifidobacterium posterior economies were prepared, which solved the problem of unbalanced nutrition and low utilization rate of unconventional fiber materials in the livestock industry, and achieved improvement Animal growth performance and the effect of reducing breeding costs.
Patent Information
- Application Number
- CN202411671871.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-21
- Publication Date
- 2025-02-25
AI Technical Summary
In the livestock industry, unconventional fiber materials have problems such as shortage of feed resources and high breeding costs due to unbalanced nutrition, anti-nutrition factors, poor palatability, and low utilization.
Using the preparation method of Bifidobacterium epibiotics, oligosaccharides, cellulase and lysozyme and fiber materials are mixed after enzymatic reaction to form a carrier, and combined with Bifidobacterium cell contents to prepare a nutrient-rich Bifidobacterium epibiotic, as feed additive.
It improves the palatability of feed, improves the daily weight gain rate of animals, reduces the feed-to-weight ratio and dead-stale food rate, reduces the breeding cost, and improves the growth performance and immune function of animals.
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of probiotics, and in particular to a bifidobacterium postbiotic and a preparation method and application thereof. Background Art
[0002] my country is a large agricultural country. In addition to major raw materials such as soybean meal and corn, there are also a large number of unconventional fiber materials, such as crop straw, cake meal, feather meal, tea residue, etc. Unconventional fiber materials are nutritionally unbalanced, contain anti-nutritional factors, and have poor palatability, but they have important potential and value in animal husbandry. Through technological innovation and comprehensive utilization, these resources can be fully utilized to alleviate the problem of feed resource shortage, reduce feed costs, improve animal production performance, and promote the sustainable development of animal husbandry.
[0003] The International Scientific Association of Probiotics and Prebiotics (ISAPP) gave a definition of postbiotics in 2021, that is, preparations of inanimate microorganisms and / or their components that are beneficial to the health of the host, including inactivated bacteria, bacteria and bacterial lysis components, metabolites, etc. As a green, stable and widely used feed additive, postbiotics can improve the growth performance of animals, increase feed utilization, thereby increasing production efficiency, and at the same time enhance the immune function of animals, regulate intestinal microecology, and improve disease resistance.
[0004] In view of this, the present invention is proposed. Summary of the invention
[0005] In order to solve the technical problems in the prior art that unconventional fiber materials are nutritionally unbalanced, contain anti-nutritional factors, have poor palatability and low utilization rate, the present invention provides a bifidobacterium postbiotic and a preparation method and application thereof. The bifidobacterium postbiotic can be used as a feed additive and added to animal feed, especially chicken feed, to improve the palatability of the feed, increase the daily weight gain rate, reduce the feed-to-weight ratio and the mortality rate, and reduce the breeding cost.
[0006] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:
[0007] One aspect of the present invention relates to a method for preparing a bifidobacterium postbiotic, comprising the following steps:
[0008] (a) adding oligosaccharides, cellulase and lysozyme to a bifidobacterium bacterial liquid to obtain a first mixed system;
[0009] (b) adding a composite reagent to a fiber material to perform a first reaction; adjusting the pH of the reaction system of the first reaction to 4 to 6, and then adding a composite enzyme preparation to perform an enzymatic hydrolysis reaction to obtain a carrier;
[0010] (c) mixing the first mixed system and the carrier to obtain bifidobacterium postbiotics.
[0011] The preparation method of the bifidobacterium postbiotics has the advantages of being simple and easy to operate. As a feed additive, the prepared postbiotics are added to animal feed, especially chicken feed, to improve the palatability of the feed, increase the daily weight gain rate, reduce the feed-to-weight ratio and the mortality rate, and reduce the breeding cost.
[0012] Another aspect of the present invention also relates to the bifidobacterium postbiotics prepared by the preparation method of the bifidobacterium postbiotics.
[0013] The bifidobacterium postbiotics have a rich variety of nutrients in sufficient quantity, which can supply the growth and development of animals, especially when added to chicken feed as a chicken feed additive, it can improve the palatability of the feed, increase the daily weight gain rate of chickens, reduce the feed-to-weight ratio and the mortality rate, and reduce the breeding cost of broilers.
[0014] Another aspect of the present invention also relates to a feed, comprising the bifidobacterium postbiotics prepared by the preparation method of the bifidobacterium postbiotics or the bifidobacterium postbiotics.
[0015] Compared with the prior art, the present invention has the following beneficial effects:
[0016] (1) The preparation method of the bifidobacterium postbiotic provided by the present invention processes the bifidobacteria to release the contents in the bifidobacterium cells to the greatest extent, thereby providing a variety and quantity of nutrients for animal growth and improving the growth performance of the animals.
[0017] (2) The preparation method of bifidobacterium postbiotics provided by the present invention uses unconventional fiber materials as raw materials for carriers, and through chemical treatment and biological treatment, converts substances in the unconventional fiber materials that are difficult to utilize into nutrients such as reducing sugars, which can be supplied to animals as nutrients for growth and development, enriching the nutritional types in postbiotics, thereby increasing the utilization rate of unconventional fiber materials and improving the growth performance of animals.
[0018] (3) The bifidobacterium postbiotics provided by the present invention include bifidobacterium cell contents and processed unconventional fiber materials, which are rich in metabolites such as organic acids, amino acids and their derivatives, oligosaccharides, small peptides, etc., and have a rich variety and sufficient quantity of nutrients, which can be supplied to the growth and development of animals, especially as a chicken feed additive added to chicken feed, which can improve the palatability of feed, increase the daily weight gain rate of chickens, reduce the feed-to-weight ratio and mortality rate, and reduce the breeding cost of broilers. DETAILED DESCRIPTION
[0019] The technical scheme of the present invention will be clearly and completely described below in conjunction with specific embodiments, but it will be appreciated by those skilled in the art that the following described embodiments are part of embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work, all belong to the scope of protection of the present invention. If specific conditions are not indicated in the embodiments, they are carried out according to normal conditions or conditions recommended by the manufacturer. If the manufacturer is not indicated in the reagents or instruments used, they are all conventional products that can be purchased commercially.
[0020] One aspect of the present invention relates to a method for preparing a bifidobacterium postbiotic, comprising the following steps:
[0021] (a) adding oligosaccharides, cellulase and lysozyme to a bifidobacterium bacterial liquid to obtain a first mixed system;
[0022] (b) adding a composite reagent to a fiber material to perform a first reaction; adjusting the pH of the reaction system of the first reaction to 4 to 6 (for example, but not limited to, any one of 4, 4.5, 5, 5.5 or 6 or a range between any two of them), and then adding a composite enzyme preparation to perform an enzymatic hydrolysis reaction to obtain a carrier;
[0023] (c) mixing the first mixed system and the carrier to obtain bifidobacterium postbiotics.
[0024] The preparation method of the bifidobacterium postbiotics processes the bifidobacteria to release the contents in the bifidobacterium cells to the maximum extent, thereby providing a variety and quantity of nutrients for animal growth and improving the growth performance of the animals.
[0025] The preparation method of the bifidobacterium postbiotics uses unconventional fiber materials as raw materials for carriers, and through chemical and biological treatments, converts substances in the unconventional fiber materials that are difficult to utilize into nutrients such as reducing sugars, which can be supplied to animals as nutrients for growth and development, enriching the nutritional types in the postbiotics, thereby increasing the utilization rate of the unconventional fiber materials and improving the growth performance of animals.
[0026] Furthermore, the OD of the bifidobacterium bacterial solution is 600 The value is 35~45.
[0027] Furthermore, the amount of oligosaccharide added to the bifidobacterium bacterial liquid is 0.5wt% to 2wt%, including but not limited to any one of 0.5wt%, 1wt%, 1.5wt% or 2wt% or any range between two of them.
[0028] Furthermore, the oligosaccharide includes but is not limited to: mannobiose and / or mannotriptose. Adding oligosaccharides to bifidobacterium postbiotics can increase the antioxidant and pathogen adsorption capabilities of the postbiotics, thereby improving the growth performance of animals, enhancing disease resistance, and reducing mortality.
[0029] Furthermore, the enzymatic activity of the cellulase is 9500-70000 U / g, including but not limited to any one of 9500 U / g, 15000 U / g, 20000 U / g, 25000 U / g, 30000 U / g, 35000 U / g, 40000 U / g, 45000 U / g, 50000 U / g, 55000 U / g, 60000 U / g, 65000 U / g or 70000 U / g or the range between any two of them.
[0030] Furthermore, the amount of cellulase added to the bifidobacterium bacterial liquid is 3wt% to 10wt%, including but not limited to any one of 3wt%, 4wt%, 5wt%, 6wt%, 7wt%, 8wt%, 9wt% or 10wt% or a range between any two of them.
[0031] Furthermore, the concentration of the lysozyme in the bifidobacterium liquid is 3 to 10 U / mL, including but not limited to any one of 3 U / mL, 4 U / mL, 5 U / mL, 6 U / mL, 7 U / mL, 8 U / mL, 9 U / mL or 10 U / mL or a range between any two of them.
[0032] Cellulase and lysozyme can break down bacterial cell walls, causing them to rupture and allow the contents of bifidobacterium cells to flow out, thereby increasing the nutritional content of postbiotics, providing richer nutrients for animal growth, and improving animal growth performance.
[0033] The present invention does not specifically limit the specific type of fiber material, and conventional fiber materials in the art can be used to implement the technical solution of the present invention. In some specific embodiments, the fiber material can be bulk fiber raw materials, and can also be unconventional fiber materials, such as crop straw, cakes, feather powder, tea residues, etc.
[0034] Furthermore, the fiber material includes, but is not limited to, rice straw and bran in a mass ratio of 75-90:10-25 (for example, but not limited to 75:25, 80:20, 85:15 or 90:10).
[0035] Furthermore, the composite reagent includes but is not limited to: sodium hyposulfite and calcium oxide in a mass ratio of 1:3 to 5 (for example, but not limited to 1:3, 1:4 or 1:5). The composite reagent chemically treats the fiber material, dissolves hemicellulose and lignin in the fiber material, allows the cellulase to fully contact the raw material, and improves the enzymolysis efficiency.
[0036] Furthermore, the addition amount of the composite agent is 3% to 6% of the mass of the fiber material, including but not limited to any point value of 3%, 4%, 5% or 6% or a range value between any two of them.
[0037] Furthermore, the temperature of the first reaction is 30-50°C, including but not limited to any one of 30°C, 35°C, 40°C, 45°C or 50°C or a range between any two of them.
[0038] Furthermore, the time of the first reaction is 6-8 hours, including but not limited to any one of 6 hours, 6.5 hours, 7 hours, 7.5 hours or 8 hours or a range between any two of them.
[0039] Furthermore, the added amount of the complex enzyme preparation is 8% to 12% of the mass of the fiber material, including but not limited to any one of 8%, 9%, 10%, 11% or 12% or a range between any two of them.
[0040] Furthermore, the composite enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 3 to 5:1 to 3:1 (for example, but not limited to 3:3:1, 4:2:1 or 5:1:1). The composite enzyme preparation can fully enzymatically hydrolyze the fiber material and convert the fiber material into nutrients that can be used for animal growth, such as the content of reducing sugar.
[0041] Furthermore, the temperature of the enzymatic hydrolysis reaction is 50-65° C., including but not limited to any point value of 50° C., 53° C., 55° C., 57° C., 59° C., 61° C., 63° C. or 65° C. or a range value between any two thereof. According to the characteristics of the composite enzyme preparation, limiting the enzymatic hydrolysis reaction temperature within a certain range can ensure that the composite enzyme preparation has good enzyme activity and fully enzymatically hydrolyzes the fiber material.
[0042] Furthermore, the enzymatic hydrolysis reaction time is 24 to 48 hours, including but not limited to any point value of 24 hours, 28 hours, 32 hours, 36 hours, 40 hours, 44 hours or 48 hours or a range value between any two of them.
[0043] Furthermore, the volume ratio of the first mixed system to the carrier is 50-70:30-50, including but not limited to 50:50, 55:65, 60:40, 65:35 or 70:30.
[0044] Furthermore, the method for preparing the bifidobacterium bacterial liquid comprises the following steps:
[0045] streak bifidobacterium onto LB solid medium to obtain a single colony; pick a single colony and inoculate it into MRS medium, and culture it at 30-37°C (for example, it can be but not limited to any one of 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C or 37°C or a range between any two) for 16-24h (for example, it can be but not limited to any one of 16h, 18h, 20h, 22h or 24h or a range between any two) to obtain a primary seed solution; transfer the primary seed solution to a secondary seed medium at an inoculum amount of 3vol%-5vol% (for example, it can be but not limited to any one of 3vol%, 3.5vol%, 4vol%, 4.5vol% or 5vol% or a range between any two), and the culture temperature is 35-37°C (for example, it can be but not limited to any one of 3vol%, 3.5vol%, 4vol%, 4.5vol% or 5vol% or a range between any two). The temperature is 35°C, 35.5°C, 36°C, 36.5°C or 37°C, or any range between them), and the culture is carried out for 8-14h (for example, it can be but not limited to any point value of 8h, 10h, 12h or 14h, or any range between them); the secondary seeds are inoculated into the tertiary seed culture medium at an inoculation amount of 10vol%-40vol% (for example, it can be but not limited to any point value of 10vol%, 15vol%, 20vol%, 25vol%, 30vol%, 35vol% or 40vol%, or any range between them), and the culture is continued for 16-24h (for example, it can be but not limited to any point value of 16h, 18h, 20h, 22h or 24h, or any range between them), to obtain a bifidobacterium culture solution.
[0046] Further, the LB medium formula is 8-10 g / L (for example, it can be but not limited to any one of 8 g / L, 9 g / L or 10 g / L or a range between any two) tryptone, 3-5 g / L (for example, it can be but not limited to any one of 3 g / L, 4 g / L or 5 g / L or a range between any two) yeast extract, 5-10 g / L (for example, it can be but not limited to any one of 5 g / L, 6 g / L, 7 g / L, 8 g / L, 9 g / L or 10 g / L or a range between any two) NaCl and 5-8 g / L (for example, it can be but not limited to any one of 5 g / L, 6 g / L, 7 g / L or 8 g / L or a range between any two) agar powder.
[0047] Further, the MRS medium formula is 8-10 g / L (for example, it can be but not limited to any one of 8 g / L, 9 g / L or 10 g / L or a range between any two) peptone, 6-8 g / L (for example, it can be but not limited to any one of 6 g / L, 7 g / L or 8 g / L or a range between any two) beef extract powder, 2-4 g / L (for example, it can be but not limited to any one of 2 g / L, 3 g / L or 4 g / L or a range between any two) yeast extract powder, 15-25 g / L (for example, it can be The present invention relates to a novel nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, which is a nanostructured carbon foam, range value between) dipotassium hydrogen phosphate (anhydrous), 2-5 g / L (for example, it can be but not limited to any one of 2 g / L, 3 g / L, 4 g / L or 5 g / L, or a range value between any two) sodium acetate, 0.5-2 g / L (for example, it can be but not limited to any one of 0.5 g / L, 1 g / L, 1.5 g / L or 2 g / L, or a range value between any two) triammonium citrate / diammonium hydrogen citrate (anhydrous), 0.05-0.2 g / L (for example, it can be but not limited to 0.05 g / L, 0.1 g / L, 0.15 g / L / L or 0.2g / L or any range between them) magnesium sulfate (anhydrous), 0.02-0.05g / L (for example, it can be but not limited to any one of 0.02g / L, 0.03g / L, 0.04g / L or 0.05g / L or any range between them) manganese sulfate, and 0.02-0.05g / L (for example, it can be but not limited to any one of 0.02g / L, 0.03g / L, 0.04g / L or 0.05g / L or any range between them) L-cysteine.
[0048] Further, the secondary seed culture medium formula is: 15-25g / L (for example, it can be but not limited to any one of 15g / L, 17g / L, 19g / L, 21g / L, 23g / L or 25g / L or a range between any two) defatted soybean meal, 3-8g / L (for example, it can be but not limited to any one of 3g / L, 4g / L, 5g / L, 6g / L, 7g / L or 8g / L or a range between any two) wheat flour, 10-15g / L (for example, it can be but not limited to any one of 10g / L, 11g / L, 12g / L, 13g / L, 14g / L or 15g / L or a range between any two) The invention relates to a pharmaceutical composition comprising the following: a) skimmed milk powder, 2-5 g / L (for example, it may be but not limited to any one of 2 g / L, 3 g / L, 4 g / L or 5 g / L or a range between two of them) isomaltooligosaccharide, 10-20 g / L glucose (for example, it may be but not limited to any one of 10 g / L, 12 g / L, 14 g / L, 16 g / L, 18 g / L or 20 g / L or a range between two of them), and 1-3 g / L (for example, it may be but not limited to any one of 1 g / L, 2 g / L or 3 g / L or a range between two of them) yeast extract, and VB1, VB2 and biotin are further added.
[0049] Furthermore, the tertiary seeds also use MRS culture medium, with 20wt% to 30wt% (for example, it can be but not limited to 20wt%, 22wt%, 24wt%, 26wt%, 28wt% or 30wt% of any one point value or any range value between two) of glucose as the feed, and ammonia water is used to adjust the pH value to 6.0 to 6.5 (for example, it can be but not limited to 6.0, 6.1, 6.2, 6.3, 6.4 or 6.5 of any one point value or any range value between two).
[0050] Another aspect of the present invention also relates to the bifidobacterium postbiotics prepared by the preparation method of the bifidobacterium postbiotics.
[0051] The bifidobacterium postbiotics include bifidobacterium cell contents and processed unconventional fiber materials, are rich in metabolites such as organic acids, amino acids and derivatives, oligosaccharides, small peptides, etc., have a rich variety and sufficient amount of nutrients, can supply the growth and development of animals, especially as a chicken feed additive added to chicken feed, can improve the palatability of feed, increase the daily weight gain rate of chickens, reduce feed-to-weight ratio and mortality rate, and reduce the breeding cost of broilers.
[0052] Another aspect of the present invention also relates to a feed, comprising the bifidobacterium postbiotics prepared by the preparation method of the bifidobacterium postbiotics or the bifidobacterium postbiotics.
[0053] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be obtained commercially.
[0054] Example 1
[0055] The preparation method of bifidobacterium postbiotics provided in this embodiment comprises the following steps:
[0056] (1) preparing a bifidobacterium bacterial liquid; streaking the bifidobacterium onto an LB solid medium to obtain a single colony; picking a single colony and inoculating it into an MRS medium, culturing it at 35° C. for 24 hours to obtain a primary seed liquid; transferring the primary seed liquid into a secondary seed medium at a 4% inoculation rate, culturing it at 35° C. for 8 hours; inoculating the secondary seed into a tertiary seed medium at a 35% inoculation rate, and continuing to culture it for 24 hours to obtain a bifidobacterium bacterial liquid;
[0057] (2) adding mannobiose, cellulase and lysozyme to the bacterial solution of bifidobacterium to obtain a first mixed system, wherein the amount of oligosaccharide added is 1 wt %; the enzymatic activity of cellulase is 10000 U / g, the amount added is 7 wt %, and the working concentration of lysozyme is 8 U / mL;
[0058] (3) Preparing a carrier: adding a composite reagent to a pulverized fiber material of rice straw and bran in a mass ratio of 90:10, performing a first reaction at 30° C. for 6 hours, adjusting the pH of the reaction system to 4.5 after the reaction, and then adding a composite enzyme preparation for sufficient enzymolysis at 55° C. to obtain a carrier;
[0059] The composite reagent includes: sodium hyposulfite and calcium oxide in a mass ratio of 1:4; the addition amount of the composite reagent is 5% of the mass of the fiber material;
[0060] The compound enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 4:1:1; the addition amount of the compound enzyme preparation is 10% of the mass of the mixed raw materials;
[0061] (4) The first mixed system and the carrier are uniformly mixed in a volume ratio of 60:40, and dried to obtain bifidobacterium postbiotics.
[0062] Example 2
[0063] The preparation method of bifidobacterium postbiotics provided in this embodiment comprises the following steps:
[0064] (1) Same as Example 1;
[0065] (2) adding mannose triglyceride, cellulase and lysozyme to the bacterial solution of bifidobacterium to obtain a first mixed system, wherein the amount of oligosaccharide added is 2 wt %; the enzymatic activity of cellulase is 9500 U / g, the amount added is 3 wt %, and the working concentration of lysozyme is 10 U / mL;
[0066] (3) Preparing a carrier: adding a composite reagent to a crushed fiber material of rice straw and bran in a mass ratio of 75:25, performing a first reaction at 30° C. for 8 h, adjusting the pH of the reaction system to 4 after the reaction, and then adding a composite enzyme preparation at 50° C. for sufficient enzymatic hydrolysis to obtain a carrier;
[0067] The composite reagent includes: sodium hyposulfite and calcium oxide in a mass ratio of 1:3; the addition amount of the composite reagent is 3% of the mass of the fiber material;
[0068] The compound enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 3:1:1; the addition amount of the compound enzyme preparation is 12% of the mass of the mixed raw materials;
[0069] (4) The first mixed system and the carrier are uniformly mixed in a volume ratio of 50:30, and dried to obtain bifidobacterium postbiotics.
[0070] Example 3
[0071] The preparation method of bifidobacterium postbiotics provided in this embodiment comprises the following steps:
[0072] (1) Same as Example 1;
[0073] (2) adding mannobiose, cellulase and lysozyme to the bacterial solution of bifidobacterium to obtain a first mixed system, wherein the amount of oligosaccharide added is 0.5 wt %; the enzymatic activity of cellulase is 70,000 U / g, the amount added is 10 wt %, and the working concentration of lysozyme is 3 U / mL;
[0074] (3) Preparing a carrier: adding a composite reagent to a crushed fiber material of rice straw and bran in a mass ratio of 80:15, performing a first reaction at 50° C. for 6 h, adjusting the pH of the reaction system to 6 after the reaction, and then adding a composite enzyme preparation for sufficient enzymatic hydrolysis at 65° C. to obtain a carrier;
[0075] The composite reagent includes: sodium hyposulfite and calcium oxide in a mass ratio of 1:5; the addition amount of the composite reagent is 6% of the mass of the fiber material;
[0076] The compound enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 5:3:1; the addition amount of the compound enzyme preparation is 8% of the mass of the mixed raw materials;
[0077] (4) The first mixed system and the carrier are uniformly mixed in a volume ratio of 70:30, and dried to obtain bifidobacterium postbiotics.
[0078] Example 4
[0079] The preparation method of bifidobacterium postbiotics provided in this embodiment comprises the following steps:
[0080] (1) Same as Example 1;
[0081] (2) adding mannose triglyceride, cellulase and lysozyme to the bacterial solution of bifidobacterium to obtain a first mixed system, wherein the amount of oligosaccharide added is 0.8 wt %; the enzymatic activity of cellulase is 20000 U / g, the amount added is 5 wt %, and the working concentration of lysozyme is 5 U / mL;
[0082] (3) Preparing a carrier: adding a composite reagent to a mixed fiber material of rice straw and bran in a mass ratio of 85:20, performing a first reaction at 35° C. for 7 h, adjusting the pH of the reaction system to 5 after the reaction, and then adding a composite enzyme preparation for sufficient enzymolysis at 60° C. to obtain a carrier;
[0083] The composite reagent includes: sodium hyposulfite and calcium oxide in a mass ratio of 1:4; the addition amount of the composite reagent is 4% of the mass of the fiber material;
[0084] The compound enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 4:3:1; the addition amount of the compound enzyme preparation is 9% of the mass of the mixed raw materials;
[0085] (4) The first mixed system and the carrier are uniformly mixed in a volume ratio of 55:45, and dried to obtain bifidobacterium postbiotics.
[0086] Example 5
[0087] The preparation method of bifidobacterium postbiotics provided in this embodiment comprises the following steps:
[0088] (1) Same as Example 1;
[0089] (2) adding mannobiose and mannotriose, cellulase and lysozyme to the bacterial solution of bifidobacterium to obtain a first mixed system, wherein the amount of oligosaccharide added is 1.7 wt %; the enzymatic activity of cellulase is 50,000 U / g, the amount added is 8 wt %, and the working concentration of lysozyme is 8 U / mL;
[0090] (3) Preparing a carrier: adding a composite reagent to a crushed fiber material of rice straw and bran in a mass ratio of 90:25, performing a first reaction at 45° C. for 8 h, adjusting the pH of the reaction system to 5.5 after the reaction, and then adding a composite enzyme preparation for sufficient enzymolysis at 53° C. to obtain a carrier;
[0091] The composite reagent includes: sodium hyposulfite and calcium oxide in a mass ratio of 1:5; the addition amount of the composite reagent is 5% of the mass of the fiber material;
[0092] The compound enzyme preparation includes: cellulase, xylanase and pectinase in a mass ratio of 5:3:1; the addition amount of the compound enzyme preparation is 11% of the mass of the mixed raw materials;
[0093] (4) The first mixed system and the carrier are uniformly mixed in a volume ratio of 65:35, and dried to obtain bifidobacterium postbiotics.
[0094] Comparative Example 1
[0095] The only difference between this comparative example and Example 1 is that in step (2), no cellulase and lysozyme are added to the bacterial solution of Bifidobacterium.
[0096] Comparative Example 2
[0097] The difference between the comparative example and Example 1 is that in step (2), lysozyme is not added to the bacterial solution of bifidobacteria.
[0098] Comparative Example 3
[0099] The difference between this comparative example and Example 1 is that the carrier in step (3) is crushed rice straw and bran in a mass ratio of 90:10.
[0100] Comparative Example 4
[0101] The difference between this comparative example and Example 1 is that in step (2), mannobiose is not added to the bifidobacterium bacterial liquid.
[0102] Comparative Example 5
[0103] The difference between this comparative example and Example 1 is that in step (3), no composite reagent is added to the fiber material.
[0104] Comparative Example 6
[0105] The difference between this comparative example and Example 1 is that in step (2), the amount of oligosaccharide added is 4 wt %.
[0106] Comparative Example 7
[0107] The difference between this comparative example and Example 1 is that in step (3), the fiber material comprises rice straw and bran in a mass ratio of 60:40.
[0108] Experimental Example 1
[0109] The components of the bifidobacterium postbiotics prepared in each embodiment and comparative example were analyzed, and the results are shown in Table 1.
[0110] Table 1
[0111]
[0112]
[0113] As can be seen from Table 2, the bifidobacterium postbiotics provided by the present invention are rich in organic acids, amino acids and their derivatives, and the content of reducing sugars is significantly increased after the carrier is treated chemically and biologically.
[0114] Experimental Example 2
[0115] The application of the bifidobacterium postbiotics prepared in each embodiment and comparative example in animal experiments, in this embodiment, broiler breeding is used as an example, and the results are shown in Table 2. The specific method is as follows:
[0116] Broilers with similar body weight and good health were selected and randomly divided into 3 groups, with 6 replicates in each group and 15 in each cage. They were fed with powder, free access to food and water, and 24 hours of light per day. The combination of artificial light and natural light was used. The experimental period was 42 days. The composition of the basic diet was: corn, soybean meal, rapeseed cake, broken rice, soybean oil, stone powder, multivitamin, multimineral and amino acids. The control group was only fed with the basic diet.
[0117] Table 2
[0118]
[0119]
[0120]
[0121]
[0122] As shown in Table 3, compared with the control group, the group supplemented with bifidobacterium significantly increased daily weight gain, reduced feed-to-meat ratio and mortality rate, and the effect increased with the increase in the amount of addition.
[0123] Although the present invention has been illustrated and described with specific embodiments, it should be appreciated that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Those skilled in the art should understand that the technical solutions described in the above embodiments may be modified, or some or all of the technical features thereof may be replaced by equivalents without departing from the spirit and scope of the present invention. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention. Therefore, this means that all such replacements and modifications within the scope of the present invention are included in the appended claims.
Claims
1. A method for preparing a bifidobacterium postbiotic, characterized in that: The following steps are involved: (a) adding oligosaccharides, cellulase and lysozyme to a bifidobacterium bacterial liquid to obtain a first mixed system; (b) adding a composite reagent to a fiber material to perform a first reaction; adjusting the pH of the reaction system of the first reaction to 4 to 6, and then adding a composite enzyme preparation to perform an enzymatic hydrolysis reaction to obtain a carrier; (c) mixing the first mixed system and the carrier to obtain bifidobacterium postbiotics.
2. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The amount of oligosaccharide added to the bifidobacterium bacterial liquid is 0.5wt% to 2wt%; And / or, the oligosaccharides include: mannobiose and / or mannotriose.
3. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The cellulase has an enzymatic activity of 9500 to 70000 U / g; And / or, the amount of cellulase added to the bifidobacterium bacterial liquid is 3wt% to 10wt%.
4. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The concentration of the lysozyme in the bifidobacterium bacterial solution is 3-10 U / mL.
5. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: Includes at least one of the following technical features: (1) The fiber material comprises: rice straw and bran in a mass ratio of 75-90:10-25; (2) The composite reagent comprises: sodium hyposulfite and calcium oxide in a mass ratio of 1:3-5; (3) The added amount of the composite reagent is 3% to 6% of the mass of the fiber material.
6. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The temperature of the first reaction is 30-50°C; And / or, the first reaction time is 6 to 8 hours.
7. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The added amount of the composite enzyme preparation is 8% to 12% of the mass of the fiber material; And / or, the composite enzyme preparation comprises: cellulase, xylanase and pectinase in a mass ratio of 3 to 5:1 to 3:
1.
8. The method for preparing bifidobacterium postbiotics according to claim 1, characterized in that: The temperature of the enzymatic hydrolysis reaction is 50-65°C; And / or, the enzymatic hydrolysis reaction time is 24 to 48 hours.
9. The bifidobacterium postbiotic prepared by the method for preparing bifidobacterium postbiotic according to any one of claims 1 to 8.
10. A feed, characterized in that: The invention comprises the bifidobacterium postbiotic prepared by the preparation method of the bifidobacterium postbiotic according to any one of claims 1 to 8 or the bifidobacterium postbiotic according to claim 9.