CN116478857B - Bifidobacterium longum variant capable of producing polysaccharide and application of polysaccharide in crystal ball probiotic preparation - Google Patents

Bifidobacterium longum variant capable of producing polysaccharide and application of polysaccharide in crystal ball probiotic preparation Download PDF

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CN116478857B
CN116478857B CN202310094263.2A CN202310094263A CN116478857B CN 116478857 B CN116478857 B CN 116478857B CN 202310094263 A CN202310094263 A CN 202310094263A CN 116478857 B CN116478857 B CN 116478857B
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coating
bifidobacterium longum
polysaccharide
probiotic
layer
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CN116478857A (en
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黄芬
田�健
全丽燕
丁慧
诸辉
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Ningbo Xinuoya Marine Biotechnology Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • 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
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides a bifidobacterium longum variant (Bifidobacterium longum subsp) capable of producing polysaccharide, wherein the bifidobacterium longum strain is preserved in the Guangdong province microorganism strain collection (GDMCC) at the year 2022 and the 11 th month, the preservation address is Guangzhou, and the preservation number is GDMCC No:63000. the bifidobacterium longum variant in the invention is easy to culture, and the yield of the polysaccharide produced and expressed by the bifidobacterium longum variant is higher.

Description

Bifidobacterium longum variant capable of producing polysaccharide and application of polysaccharide in crystal ball probiotic preparation
Technical Field
The invention relates to the technical field of microorganisms, in particular to a bifidobacterium longum variant capable of producing polysaccharide and application thereof.
Background
Bifidobacterium longum (Bifidobacterium longum subsp) is a gram-positive, spore-free, motile-free, catalase-negative anaerobic bacterium. In taxonomies bifidobacteria belong to the class actinomycetes, the heterolactic fermentation, the fermentation end products being mainly acetic acid and lactic acid. Bifidobacteria are bacteria that are frequently present in the human intestinal tract and represent about 4.4-6.9% of the total bacterial count in the human intestinal tract. Bifidobacteria are beneficial to human body, such as inhibiting the growth of pathogenic microorganisms, regulating intestinal flora balance, resisting tumor and anticancer activity, treating diarrhea, enhancing body immunity, reducing cholesterol, etc.
Polysaccharides are non-starch polysaccharides formed by connecting D-glucose with a glucosidic bond, and are widely found in nature. The polysaccharide plays an important role in promoting health and preventing diseases, has positive effects in controlling postprandial blood sugar and weakening insulin response, reducing cholesterol and hyperlipidemia, enhancing organism immunity, protecting intestinal brain health and the like, and has great potential for developing functional foods, medical care, food additives and other health industry fields.
Probiotics must pass through the gastric environment to reach the intestinal tract in large amounts of viable bacteria and colonize the intestinal mucosa to exert their physiological functions. In order to ensure the survival rate of the probiotics and maintain the physiological activity of the probiotics and improve the storage stability of the probiotics in the shelf life of products, more research is currently being carried out at home and abroad on microcapsule embedding technology of the probiotics. In the embedding technology, the survival rate and stability of probiotics mainly depend on the concentration and types of thalli, the types of microcapsule wall materials, the outlet temperature of a spray dryer and the like, the influence factors are more, the preparation process is complex, three to four layers or more often need to be embedded, and the death of thalli in the embedding process cannot be effectively avoided, so that the number of viable probiotics of a final product is very low and the production cost is high.
Therefore, there is an urgent need to find a pellet probiotic preparation which has a simple coating process, good storage stability and high survival rate of probiotics, can reach the intestinal tract through the survival of strong acid gastric juice, and improves the health of the intestinal tract.
Disclosure of Invention
One of the technical problems to be solved by the invention is to provide a bifidobacterium longum (Bifidobacterium longum subsp) capable of producing polysaccharide, wherein the bifidobacterium longum strain is preserved in the Guangdong province microorganism strain preservation center (GDMCC) at the year of 2022 and the preservation number is GDMCC No:63000.
alternatively, the bifidobacterium longum DNA sequence is SEQ ID No.1.
Alternatively, the bifidobacterium longum is isolated from sea water in the beach of the south China sea.
Alternatively, the bifidobacterium longum expresses a polysaccharide yield of 0.4-0.6%.
The second technical problem to be solved by the invention is to provide an application of polysaccharide expressed by bifidobacterium longum in a crystal ball probiotic preparation.
In order to solve the problems, the invention aims to provide a three-layer coated pellet probiotic preparation, which comprises a first grease coating layer for pellets of pellet probiotics, a second isolation coating layer for coating the pellets of probiotics and a third enteric coating layer for coating the isolation layer from inside to outside; the preparation method comprises extruding, rounding and embedding probiotics with slow release adjuvants, spraying oil to form a first coating layer to obtain pellet, using polysaccharide solution produced and expressed by Bifidobacterium longum variant as coating solution, performing second layer isolation coating, sieving, using intestinal solution to perform third layer coating, and sieving to obtain the final product.
The invention is realized by the following technical scheme:
the three-layer coated pellet probiotic preparation comprises a first grease coating layer of pellet probiotic pellets, a second isolation coating layer for wrapping the probiotic pellets and a third enteric coating layer for wrapping the isolation layer from inside to outside; the crystal ball probiotic micro-pellets are prepared by extruding and rounding probiotic concentrated solution and/or probiotic powder and food-grade auxiliary materials thereof, and then spraying grease; the second layer of isolation coating is made of polysaccharide material; the third enteric coating is an enteric material.
The probiotics comprise beneficial bacteria and/or beneficial fungi, wherein the beneficial bacteria are selected from any one or a combination of at least two of bifidobacterium, lactobacillus mucilaginosus, lactobacillus, streptococcus, lactococcus, propionibacterium, leuconostoc, pediococcus, weizmann bacteria, zoococcus and staphylococcus; the beneficial fungus is selected from any one or a combination of at least two of Bifidobacterium, lactobacillus casei, lactobacillus mucilaginosus, lactobacillus guangdaliensis, streptococcus, lactococcus, propionibacterium, leuconostoc, pediococcus, weizmann, pediococcus, staphylococcus, kluyveromyces, saccharomyces cerevisiae, brevibacterium, monascus purpureus, or Monascus purpureus.
The preparation method of the crystal ball probiotic preparation comprises the following steps:
(1) Dissolving the polysaccharide material in water to prepare a solution with the mass concentration of 1-15%, taking the solution as a second coating solution, coating the solution by using a fluidized bed, and sieving the solution to obtain a second coating;
(2) The enteric-coated material is prepared into a solution with the mass percentage concentration of 5-20% by using ethanol solution or absolute ethanol with the volume concentration of 60-85% as a third coating solution, and the third coating is obtained by coating by using a fluidized bed and sieving.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the invention separates and identifies a strain of polysaccharide-producing bifidobacterium longum (Bifidobacterium longum subsp), more particularly a strain of polysaccharide-producing bifidobacterium hainanensis variant. The strain is determined to be bifidobacterium longum by genome sequence analysis and construction of phylogenetic tree.
2. Compared with the prior art, the bifidobacterium longum is easy to culture, and the yield and activity of the polysaccharide produced and expressed by the bifidobacterium longum are higher.
3. The crystal ball probiotics have good storage stability, can keep activity in the normal temperature storage process, and effectively solve the problem that the number of viable probiotics is greatly reduced in the common processing and selling links of the probiotic products;
4. the survival rate of probiotics is high, and the probiotics are not easy to inactivate in gastric juice and bile;
5. the probiotics can be slowly released in the intestinal tract and can be smoothly planted in the intestinal tract;
6. the operation is simple and the process is easy to control.
Drawings
FIG. 1 is a colony chart of strain XNY6069 in example 1 of the present invention;
FIG. 2 is a microscopic image of strain XNY6069 in example 1 of the present invention;
FIG. 3 is an agarose gel electrophoresis chart of PCR amplified products of strain XNY6069 in example 2 of the present invention;
FIG. 4 is a phylogenetic tree of strain XNY6069 in example 2 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples, for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, and the description thereof is merely illustrative of the present invention and not intended to be limiting.
EXAMPLE 1 acquisition of Bifidobacterium longum (Bifidobacterium longum subsp) Strain
1. Preparation of culture Medium
Culture medium (g/L): 10.0 parts of beef extract, 5.0 parts of peptone, 3.0 parts of yeast powder, 5.0 parts of D (+) -glucose, 1.0 parts of soluble starch, 5.0 parts of sodium chloride, 3.0 parts of anhydrous sodium acetate and 0.5 parts of L-cysteine hydrochloride
2. Screening of strains
Collecting some seawater samples from the coastal beach of Hainan, culturing the collected samples one by one, coating the samples in MRS flat plate culture medium after gradient dilution, numbering corresponding flat plates respectively, placing the flat plates in a constant temperature incubator at 37 ℃ for anaerobic culture for 24 hours, selecting single bacterial colonies with better growth vigor, and respectively inoculating the single bacterial colonies to test tubes for secondary culture for bacterial preservation. The colony profile was apparent and the color was off-white (see FIGS. 1 and 2).
EXAMPLE 2 Gene sequence analysis of Bifidobacterium longum (Bifidobacterium longum subsp) Strain
1. Fungal DNA genome extraction:
the strain broth obtained in example 1 was centrifuged, and the precipitate was extracted by CTAB method to obtain 16 srDNA.
2. PCR amplification of 16S rDNA was performed on this strain:
and (3) PCR amplification: primers 515F and 806R were PCR amplified, 515F was 5'-GTGYCAGCMGCCGCGGTAA-3', and 806R was 5'-GGACTACNVGGGTWTCTAAT-3'. The PCR reaction system is as follows: 25. Mu.L of TAQ enzyme, 2. Mu.L of 515F primer, 2. Mu.L of 806R primer, 5. Mu.L of DNA template and 16. Mu.L of sterile water. The PCR amplification procedure was: pre-denaturation at 94℃for 5min; denaturation at 94℃for 60s, annealing at 48℃for 60s, extension at 72℃for 60s,28 cycles; the reaction was terminated by extension at 72℃for 7min and at 4 ℃. Gel electrophoresis and PCR product gel recovery: the PCR products were identified by 1.5% agarose gel electrophoresis. The target PCR product was recovered and purified using a rapid agarose gel DNA recovery kit (Beijing Baitaike Biotechnology Co., ltd.) and subjected to 16S rDNA sequencing. The PCR products were identified by 1% agarose gel electrophoresis (see FIG. 3). It was observed whether or not the target band was amplified.
3.16S rDNA sequencing and analysis
The target PCR product is recovered and purified by using a rapid agarose gel DNA recovery kit (Beijing Baitaike Biotechnology Co., ltd.), and the purified PCR product is sent to Shanghai Biotechnology engineering service Co., ltd for sequencing, and the gene sequence is shown as SEQ ID NO: 1. The measured sequence was submitted to GENBANK from NCBI, U.S. to obtain a similar sequence, which was analyzed by BLAST tools and DNAMAN software for alignment and phylogenetic tree construction using the Neighbor-Joining method, as shown in FIG. 4.
The similarity of the strain to Bifidobacterium longum subsp is found to be 98.38% higher by phylogenetic tree, so that the strain can be determined as a marine bifidobacterium longum, which we named Bifidobacterium longum subsp XNY6069.
The bifidobacterium longum strain obtained by the screening is named as bifidobacterium longum strain (Bifidobacterium longum subsp), the strain number is XNY6069, the 2022 is 11 and 28 days, and the bifidobacterium longum strain is preserved in Guangdong province microorganism strain preservation center (GDMCC), the preservation address is Guangzhou, and the preservation number is GDMCC No:63000.
EXAMPLE 3 yield of polysaccharide expressed by Bifidobacterium longum (Bifidobacterium longum subsp) Strain
Polysaccharide yield determination method: centrifuging bacterial liquid, and weighing supernatant fluid M; then adding absolute ethyl alcohol with the mass 2 times of that of the supernatant, stirring and dehydrating by a glass rod for 10min, precipitating by alcohol, filtering floccules, putting the floccules on dried filter paper, and then adding a prepared ethanol solution for washing and filtering; and (5) after the filtration is finished, placing the mixture into an oven, baking the mixture to constant weight, and weighing the mass m. The calculated polysaccharide yield was M/m=0.4%.
Example 4 this example provides a pellet extrusion spheronization first layer oil coating preparation method as follows:
(1) Uniformly mixing 36g of microcrystalline cellulose, 52.8g of sodium alginate, 36g of lactose and 48g of streptococcus acidophilus with 3000 hundred million cfu/g;
(2) 120g of pure water was mixed with 7.2g of calcium chloride to form a 6% aqueous solution of calcium chloride, which was used as a wetting agent;
(3) Slowly adding the component (2) into the component (1), and uniformly mixing;
(4) The above mixture was extruded at a speed of 10rmp and then rounded in a spheronizer for 5 minutes at 1500 rmp;
(5) Drying by using a low-temperature vacuum drying oven until the moisture content of the material is less than or equal to 5%;
(6) The first layer of grease coating is carried out by a fluidized bed, the used grease is any one or the combination of at least two of solid palm oil and fish oil, the softening point of the grease is 45-60 ℃, a spray gun is provided with a heating and heat-preserving device to ensure the normal atomization of the grease, the atomization pressure is 2bar, the blowing frequency is 30Hz, the heating temperature is 60 ℃, the material temperature is 36 ℃, the particles in the bed continuously flow to form a good fluidization state, the weight of the coating is increased by 1-2%, and the fluidized coating is sieved by a sieve of 20-40 meshes.
Example 5 this example provides a pellet extrusion spheronization first layer oil coating preparation method as follows:
(1) Weighing 100g2000 hundred million cfu/g lactobacillus rhamnosus, 80g1000 hundred million cfu/g bifidobacterium lactis, 30g palm oil, 9.6g magnesium stearate and 7.2g microcrystalline cellulose, and uniformly mixing;
(2) Weighing 60g of pure water as a wetting agent;
(3) Slowly adding the component (2) into the component (1), and uniformly mixing;
(4) The above mixture was extruded at a speed of 10rmp and then rounded in a spheronizer for 5 minutes at 1500 rmp;
(5) Drying by using a low-temperature vacuum drying oven until the moisture content of the material is less than or equal to 5%;
(6) The first layer of grease coating is carried out by a fluidized bed, the used grease is any one or the combination of at least two of solid palm oil and fish oil, the softening point of the grease is 45-60 ℃, a spray gun is provided with a heating and heat-preserving device to ensure the normal atomization of the grease, the atomization pressure is 2bar, the blowing frequency is 30Hz, the heating temperature is 60 ℃, the material temperature is 36 ℃, the particles in the bed continuously flow to form a good fluidization state, the weight of the coating is increased by 1-2%, and the fluidized coating is sieved by a sieve of 20-40 meshes.
Example 6 this example provides a pellet extrusion spheronization first layer oil coating preparation method as follows:
(1) Weighing 222g6000 hundred million cfu/g of bifidobacterium animalis, 31g of solid fish oil, 15g of magnesium stearate and 31g of erythritol, and uniformly mixing;
(2) 180g of pure water was weighed as a wetting agent;
(3) Slowly adding the component (2) into the component (1), and uniformly mixing;
(4) The above mixture was extruded at a speed of 10rmp and then rounded in a spheronizer for 5 minutes at 1500 rmp;
(5) Drying by using a low-temperature vacuum drying oven until the moisture content of the material is less than or equal to 5%;
(6) The first layer of grease coating is carried out by a fluidized bed, the used grease is any one or the combination of at least two of solid palm oil and fish oil, the softening point of the grease is 45-60 ℃, a spray gun is provided with a heating and heat-preserving device to ensure the normal atomization of the grease, the atomization pressure is 2bar, the blowing frequency is 30Hz, the heating temperature is 60 ℃, the material temperature is 36 ℃, the particles in the bed continuously flow to form a good fluidization state, the weight of the coating is increased by 1-2%, and the fluidized coating is sieved by a sieve of 20-40 meshes.
The viable count in the pellet probiotic pellets of examples 4 to 6 was measured and the survival rate calculated according to GB 4789.35-2016 food safety national Standard food microbiology test lactic acid bacteria test, respectively, and the specific test results are shown in Table 1:
table 1 table of viable count of pellet probiotics of examples 4 to 6
As can be seen from Table 1, the survival rate of the viable count of the pellet in example 6 is highest and reaches more than 90%, so that the pellet in example 6 is selected for the subsequent coating process.
Example 7
1. And (2) coating a second layer: 10g of pullulan is added into 190g of hot water with the temperature of 90 ℃ and uniformly stirred at constant temperature to prepare a second layer of coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 8
1. And (2) coating a second layer: 10g of soybean polysaccharide is added into 190g of hot water with the temperature of 90 ℃ and uniformly stirred at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 9
1. And (2) coating a second layer: 10g of pectin is added into 190g of hot water at 90 ℃ and stirred uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 10
1. And (2) coating a second layer: firstly, adding 6g of citric acid into 190g of hot water at 90 ℃, then adding 4g of chitosan into the mixture, and uniformly stirring the mixture at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 11
1. And (2) coating a second layer: adding 5g of polysaccharide expressed by bifidobacterium longum into 195g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 12
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 13
1. And (2) coating a second layer: 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of soybean polysaccharide are added into 142.5g of hot water at 90 ℃ and stirred uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 14
1. And (2) coating a second layer: 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pectin are added into 142.5g of hot water at 90 ℃ and stirred uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 15
1. And (2) coating a second layer: firstly, adding 6g of citric acid into 142.5g of hot water at 90 ℃, adding 5g of polysaccharide expressed by bifidobacterium longum, then adding 2.5g of chitosan, and uniformly stirring at constant temperature to prepare a second layer of coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, adding 80g of absolute ethyl alcohol into 20g of lac, stirring and dissolving the lac into transparent liquid at normal temperature, and adding 2g of glycerol to prepare a third layer coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 16
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: firstly, 10g of zein is added with 90g of 80% ethanol, stirred and dissolved at 60 ℃ to form transparent liquid, and 1g of glycerol is added to prepare a third coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 17
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: 10g of polyvinyl acetate phthalate is firstly added with 90g of absolute ethyl alcohol, stirred and dissolved at 60 ℃ to form transparent liquid, and 1g of glycerol is added to prepare a third coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 18
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: 10g of hydroxypropyl methylcellulose phthalate is firstly taken, 90g of absolute ethyl alcohol is added, stirring and dissolving are carried out at 60 ℃ to obtain transparent liquid, and 1g of glycerol is added to prepare the third coating solution. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 19
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: the third coating solution was prepared by commercially available opadry. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 20
1. And (2) coating a second layer: and adding 5g of polysaccharide expressed by bifidobacterium longum and 2.5g of pullulan into 142.5g of hot water at 90 ℃ and stirring uniformly at constant temperature to prepare a second coating solution. Coating by using a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 56 ℃, the material temperature is 36 ℃, and the second layer of coated crystal balls are obtained by sieving with a 20-40 mesh sieve.
2. And (3) coating a third layer: commercially availableA third coating solution is prepared. Coating by a fluidized bed, wherein the atomization pressure is 2.5bar, the heating temperature is 50 ℃, the material temperature is 34 ℃, and the mixture is sieved by a sieve with 20 meshes to 40 meshes to obtain the crystal ball probiotic preparation.
Example 21 Effect of coating films made from different coating materials on preparation of pellet probiotic formulations
The prepared probiotic preparation is respectively detected in low temperature, normal temperature storage stability, survival rate in gastric juice, survival rate in bile and survival rate in intestinal juice.
The number of viable probiotic bacteria in the probiotic preparation was detected according to GB 4789.35-2016 food safety national Standard food microbiology test lactic acid bacteria test, and the survival rate was calculated.
The method for detecting the storage stability at low temperature and normal temperature comprises the following steps: respectively storing at-4deg.C and 25deg.C for 3 months, and detecting survival rate of probiotics; the method for detecting the survival rate in gastric juice comprises the following steps: preparing artificial gastric juice (containing 3.2mg/ml pepsin and 0.2% (w/v) NaCl solution) according to a pharmacopoeia method, wherein the pH is 1.2, putting the probiotic preparation into an artificial gastric juice environment at 37 ℃, and detecting the survival rate of probiotics after 2 hours; the detection method of the survival rate in bile comprises the following steps: preparing artificial simulated cholic acid (2% (w/v) pig bile extract) according to a pharmacopoeia method, putting a probiotic preparation into a simulated cholic acid environment at 37 ℃, and detecting the survival rate of probiotics after 2 hours; the method for detecting the survival rate in intestinal juice comprises the following steps: preparing artificial simulated intestinal juice (containing 1% (w/v) pancreatin) according to a pharmacopoeia method, putting the probiotic preparation into a simulated intestinal juice environment at 37 ℃, and detecting the survival rate of the probiotics after 40 minutes; the calculation formula of the survival rate: survival (%) = (N/N0) ×100, N is the number of plated colonies after treatment, N0 is the number of plated colonies before treatment, and the number of colonies is obtained: about 1g of the collected probiotic preparation is added into 1000mL of phosphate buffer solution (pH=6.8-7.0), dispersed for 5min by a homogenizer, a proper amount of slurry is taken, diluted by a certain multiple and coated on MRS agar medium, and all colony numbers on the plate are counted after culturing for 72+/-2 h at 36 ℃. The test results are shown in Table 2.
TABLE 2 influence of coating films prepared from different single polysaccharide coatings on preparation of pellet probiotic formulations
As can be seen from table 2, due to the different polysaccharide types, when the third enteric coating material is the same and the second layer is coated with a single polysaccharide, only the probiotic preparation prepared by using the polysaccharide coating expressed by bifidobacterium longum can obtain better low temperature, normal temperature stability, better gastric juice survival rate and higher survival rate in intestinal tract. Therefore, the polysaccharide expressed by bifidobacterium longum is most suitable as a probiotic sugar coating material.
TABLE 3 influence of coating films prepared from different polysaccharides on preparation of pellet probiotic formulations
Sample of Survival rate at-4℃% Survival rate at 25℃% Survival rate in gastric juice% Survival rate in bile% Survival rate in intestinal juice%
Example 12 99.16 98.82 96.17 95.21 88.87
Example 13 94.51 89.68 87.52 84.23 77.61
Example 14 93.86 88.91 86.71 85.42 78.12
Example 15 95.27 90.35 88.19 85.64 79.18
As can be seen from Table 3, when the polysaccharide expressed by Bifidobacterium longum and other commercially available conventional polysaccharide compound coatings are adopted, the probiotic pellet coated with the polysaccharide-pullulan compound coating expressed by Bifidobacterium longum has better coating effect, better low-temperature and normal-temperature stability, and very high survival rate in gastric juice, bile and intestinal juice.
As can be seen from comparison of examples 7, 11 and 3 and example 12 in Table 2, the polysaccharide expressed by bifidobacterium longum alone or the pullulan polysaccharide alone is used for coating the polysaccharide which is not expressed by bifidobacterium longum-pullulan compound coating effect is good, which indicates that the polysaccharide expressed by bifidobacterium longum and the pullulan can play a synergistic effect, and the performance of the crystal ball probiotics is further improved. The membrane structure is more compact and can better resist the external severe environment due to the fact that the pullulan polysaccharide and the polysaccharide expressed by bifidobacterium longum are crosslinked into a membrane through a hydrogen bond interpenetrating network.
In addition, the effect of the polysaccharide expressed by the bifidobacterium longum and the compound coating of the soybean polysaccharide, the pectin and the chitosan are respectively compared, and researches prove that the polysaccharide expressed by the bifidobacterium longum has no obvious synergistic effect when being compounded with the soybean polysaccharide, the pectin and the chitosan.
TABLE 4 influence of coating films made of different enteric materials on preparation of pellet probiotic formulations
Sample of Survival rate at-4℃% Survival rate at 25℃% Survival rate in gastric juice% Survival rate in bile% Survival rate in intestinal juice%
Example 16 99.21 98.53 96.28 95.37 88.58
Example 17 99.42 98.61 96.42 95.81 88.24
Example 18 99.35 98.49 96.37 95.73 88.39
Example 19 99.43 98.27 96.48 95.68 88.41
Example 20 99.37 98.39 96.51 95.54 88.62
As can be seen from Table 4, when the second layer adopts the polysaccharide expressed by Bifidobacterium longum-pullulan compound coating and the third layer adopts different enteric coating materials, the coating effect is not very different, indicating that the coating is prepared from different enteric materials
The coating film has no influence on the preparation of the crystal ball probiotic preparation, namely, when the second layer adopts polysaccharide expressed by bifidobacterium longum-pullulan to be coated in a compounding way and then the third layer is coated in an enteric coating way, the enteric coating material matched with the second layer is not needed to be selected, so that the universality is strong, the simplicity and convenience are realized, and the applicability is wide.
Although the present invention is disclosed above, the present invention is not limited thereto. For example, the application range of the composition in medicine and food can be expanded. Various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention, and the scope of the invention should be assessed accordingly to that of the appended claims.
Sequence(s)
SEQ ID NO：1
TAGGGAAATGTGGGGGCGTGCCTTACCATTGCAAGTCGAACGGGATCCATCGGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGTTCCAGTTGATCGCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGTAACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATACGGCCCCTACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGTGAGTGAGTTTACCCGTTGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTATATATCGGGAAGAACACCAATGGCGAAGGCATGTCTCTGGACCGTTACTGACGCTGAGGAGCGAAAAGCGTGGGGAGCGAAAAAGGATTAGATACCCTGGTAGTCCACGCCCGCAGACGGT。

Claims (5)

1. A bifidobacterium longum XNY6069 strain capable of producing polysaccharide, which is characterized in that: the bifidobacterium longum strain was deposited at the collection of microbiological strains (GDMCC) in the cantonese province at 12 months 22 of 2022 under the deposit number GDMCC No:63000; the bifidobacterium longum is separated from sea water in the beach of the south China sea.
2. A method for preparing a three-layer coated crystal ball probiotic preparation, which is characterized by comprising the following steps: the capsule comprises a first grease coating layer of the pellet, a second isolation coating layer for wrapping the pellet, and a third enteric coating layer for wrapping the isolation layer from inside to outside; the crystal ball probiotic micro-pellets are prepared from probiotic concentrated solution and/or probiotic powder and food-grade auxiliary materials through extrusion and spheronization;
the first grease coating layer comprises any one or the combination of two of solid palm oil and solid fish oil; the second barrier coating comprises a polysaccharide expressed by a strain of bifidobacterium longum according to claim 1 or a combination of a polysaccharide expressed by a strain of bifidobacterium longum according to claim 1 and pullulan; the third enteric coating is made of enteric materials and comprises one or more of shellac, alcohol-soluble zein, fibroin, acrylic resin and derivatives thereof, cellulose and derivatives thereof, and polyvinyl alcohol derivatives.
3. A method of preparing a three-layer coated pellet probiotic formulation according to claim 2, wherein: the food-grade auxiliary materials comprise any one or a combination of at least two of celluloses, saccharides, sugar alcohols, starches, natural colloids, animal and vegetable proteins, grease, animal and vegetable waxes and alginates; the natural colloid comprises guar gum, locust bean gum, flaxseed gum, acacia gum, tragacanth gum, ghatti gum, karaya gum, peach gum, pectin, konjac gum, gelatin, fish gelatin, xanthan gum, gellan gum, agar, carrageenan, and/or red algae gum.
4. The method for preparing the pellet probiotic preparation according to claim 2, comprising the following steps:
(1) Dissolving the polysaccharide material in water to prepare a solution with the mass concentration of 1-15%, taking the solution as a second coating solution, coating the solution by using a fluidized bed, and sieving the solution to obtain a second isolation coating;
(2) The enteric coating material is prepared into a solution with the mass percentage concentration of 5-20% by using ethanol solution or absolute ethanol with the volume concentration of 60-85% as a third coating solution, and the third enteric coating is obtained by coating and sieving a fluidized bed.
5. The use of a polysaccharide expressed by a strain of bifidobacterium longum according to claim 1, characterized in that: the polysaccharide expressed by the bifidobacterium longum is used for preparing a three-layer coated crystal ball probiotic preparation; the polysaccharide is used for a three-layer coated pellet probiotic preparation, and comprises a first grease coating layer of pellet probiotic pellets, a second isolation coating layer used for coating the probiotic pellets and a third enteric coating layer used for coating the isolation layer from inside to outside; the crystal ball probiotic micro-pellets are prepared from probiotic concentrated solution and/or probiotic powder and food-grade auxiliary materials through extrusion and spheronization;
the first grease coating layer comprises any one or the combination of two of solid palm oil and solid fish oil; the second barrier coating comprises a polysaccharide expressed by a strain of bifidobacterium longum according to claim 1 or a combination of a polysaccharide expressed by a strain of bifidobacterium longum according to claim 1 and pullulan; the third enteric coating is made of enteric materials and comprises one or more of shellac, alcohol-soluble zein, fibroin, acrylic resin and derivatives thereof, cellulose and derivatives thereof, and polyvinyl alcohol derivatives.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010168309A (en) * 2009-01-22 2010-08-05 Kaneka Corp Particulate composition and method for producing the same
CN104894005A (en) * 2015-04-30 2015-09-09 江苏紫石微康生物科技有限公司 Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof
WO2016200048A1 (en) * 2015-06-11 2016-12-15 주식회사 종근당바이오 Method for increasing survival rate, storage stability, acid resistance or bile resistance of lactic acid bacterium
WO2017191093A1 (en) * 2016-05-02 2017-11-09 Lacto Research Sprl Method for coating microorganisms, powder of said coated microorganisms, and pharmaceutical, nutraceutical, cosmetic, food or sanitary composition comprising said powder
CN113293113A (en) * 2021-06-21 2021-08-24 美益添生物医药(武汉)有限公司 Bifidobacterium longum MI-186 and application thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050266069A1 (en) * 2002-09-06 2005-12-01 Simmons Donald L Stable probiotic microsphere compositions and their methods of preparation
US20040103821A1 (en) * 2002-11-29 2004-06-03 Freund Industrial Co., Ltd. Aqueous shellac coating agent and production process therefor, and coated food and production process therefor, coated drug and production process therefor, glazing composition for oil-based confectionary, glazing process, and glazed oil-based confectionary using same
CN111134333A (en) * 2018-11-06 2020-05-12 内蒙古蒙牛乳业（集团）股份有限公司 Food or health product containing probiotic microcapsules and preparation method thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010168309A (en) * 2009-01-22 2010-08-05 Kaneka Corp Particulate composition and method for producing the same
CN104894005A (en) * 2015-04-30 2015-09-09 江苏紫石微康生物科技有限公司 Extracellular polysaccharide high-yielding lactic acid bacterium BL21, and preparation method and application thereof
WO2016200048A1 (en) * 2015-06-11 2016-12-15 주식회사 종근당바이오 Method for increasing survival rate, storage stability, acid resistance or bile resistance of lactic acid bacterium
WO2017191093A1 (en) * 2016-05-02 2017-11-09 Lacto Research Sprl Method for coating microorganisms, powder of said coated microorganisms, and pharmaceutical, nutraceutical, cosmetic, food or sanitary composition comprising said powder
CN113293113A (en) * 2021-06-21 2021-08-24 美益添生物医药(武汉)有限公司 Bifidobacterium longum MI-186 and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
长双歧杆菌BBMN68寡糖利用预测与验证;欧扬雯珊 等;《中国食品学报》;第20卷(第1期);摘要 *

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