CN111671090A - Probiotic composition and preparation thereof - Google Patents

Probiotic composition and preparation thereof Download PDF

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Publication number
CN111671090A
CN111671090A CN202010456985.4A CN202010456985A CN111671090A CN 111671090 A CN111671090 A CN 111671090A CN 202010456985 A CN202010456985 A CN 202010456985A CN 111671090 A CN111671090 A CN 111671090A
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parts
lactobacillus
fermentation culture
vitamin
probiotic
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王中振
谢涛
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Yan Ruyu Pharmaceutical Technology Co ltd
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Yan Ruyu Pharmaceutical Technology Co ltd
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    • 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
    • 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
    • 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
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • 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
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A23L33/24—Cellulose or derivatives thereof
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • 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
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/123—Bulgaricus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/167—Pentosus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/181—Salivarius
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus

Abstract

The invention relates to the technical field of probiotics, in particular to a probiotic composition and a preparation thereof, wherein the probiotic composition comprises the following raw materials in parts by weight: 0.5-4 parts of streptococcus thermophilus, 0.5-2 parts of lactobacillus acidophilus, 0.5-2.5 parts of lactobacillus bulgaricus, 0.5-2 parts of bifidobacterium lactis, 0.5-1.5 parts of micrococcus krusei, 1.5-2.5 parts of bacillus subtilis, 0.5-1.5 parts of lactobacillus pentosus, 0.5-1.2 parts of lactobacillus salivarius, 2-4 parts of bacillus cereus, 0.5-1 part of saccharomycete and 6-10 parts of protective agent. The probiotic composition has high viable bacteria survival rate, and can promote intestinal digestion and absorption, and effectively regulate intestinal flora ecosystem balance.

Description

Probiotic composition and preparation thereof
Technical Field
The invention relates to the technical field of probiotics, in particular to a probiotic composition and a preparation thereof.
Background
Probiotics are bacteria that produce lactic acid when carbohydrates such as glucose are utilized. Probiotics share the common feature of growing both aerobically and anaerobically, i.e., they are mostly facultative anaerobes. The probiotics has close relationship with human, is the most important probiotic group in human intestinal tracts, and has important effects on nutrition, immunity, health care, even anti-aging and longevity of human bodies. The higher the proportion of probiotics in the human intestinal tract, the better the micro-ecological environment and the more young and active the intestinal tract.
The probiotics are roughly divided into three groups, including lactobacillus (such as lactobacillus acidophilus, lactobacillus casei, lactobacillus jensenii, lactobacillus raman and the like), bifidobacterium (such as bifidobacterium longum, bifidobacterium breve, bifidobacterium ovorans, bifidobacterium thermophilum and the like), gram-positive coccus (such as streptococcus faecalis, lactococcus lactis, streptococcus intermedius and the like), and the probiotics have the following functions in combination: (1) improving immunity: a large amount of immunoglobulin is generated to activate immune cells of an organism, so that external bacteria are difficult to remain in the body, and the immunoregulation effect is achieved; (2) improving the gastrointestinal tract function: a protective barrier is formed on the surface of the intestinal mucosa to resist the invasion and damage of harmful bacteria to the intestinal mucosa, so that an environment which is not beneficial to the growth of the harmful bacteria is created, and the growth and the propagation of the harmful bacteria are inhibited; (3) the nutrition function is as follows: lactic acid and acetic acid are produced by fermentation, so that the utilization rate of calcium, phosphorus and iron can be improved, and digestion and absorption of related nutrient substances are promoted; (4) bowel relaxing and moistening: can generate a large amount of acidic substances in the intestines to stimulate the intestinal peristalsis and play a role in relaxing the bowels. Therefore, research and development on probiotics and related preparations can bring positive promotion significance to human health.
Patent application CN105641209A discloses a probiotic composition for improving immunity, which comprises probiotic dry powder, astragalus extract powder and medlar extract powder supplemented with nutritional ingredients such as vitamin K, vitamin B12, pyridoxine, biotin, folic acid, nicotinic acid and thiamine, and has the advantages of more ingredients, high raw material cost, and probiotic bacteria in living bacteria, easy loss of activity in transportation and storage processes, high acidity and high permeability in gastrointestinal tract and environment containing various hydrolytic enzymes, and great threat to probiotic bacteria, easy inactivation, difficulty in having enough living bacteria to reach intestinal tract or definite value in intestinal tract to play a probiotic role, and thus, space for improvement is provided.
Disclosure of Invention
Aiming at the defects in the prior art, one of the purposes of the invention is to provide a probiotic composition which has high live bacteria survival rate in the transportation and storage processes, can promote the digestion and absorption of intestinal tracts and effectively regulates the ecological system balance of intestinal flora.
The above object of the present invention is achieved by the following technical solutions:
the probiotic composition comprises the following raw materials in parts by weight: 0.5-4 parts of streptococcus thermophilus, 0.5-2 parts of lactobacillus acidophilus, 0.5-2.5 parts of lactobacillus bulgaricus, 0.5-2 parts of bifidobacterium lactis, 0.5-1.5 parts of micrococcus krusei, 1.5-2.5 parts of bacillus subtilis, 0.5-1.5 parts of lactobacillus pentosus, 0.5-1.2 parts of lactobacillus salivarius, 2-4 parts of bacillus cereus, 0.5-1 part of saccharomycete and 6-10 parts of protective agent.
Further, the feed comprises the following raw materials in parts by weight: 0.5-2 parts of streptococcus thermophilus, 1-2 parts of lactobacillus acidophilus, 1-2.5 parts of lactobacillus bulgaricus, 1-2 parts of bifidobacterium lactis, 0.5-1 part of micrococcus krusei, 2-2.5 parts of bacillus subtilis, 0.5-1.5 parts of lactobacillus pentosus, 0.5-1.0 part of lactobacillus salivarius, 2-3 parts of bacillus cereus, 0.5-1 part of saccharomycete and 6-9 parts of a protective agent.
Further, the feed comprises the following raw materials in parts by weight: 1.8 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.6 parts of bifidobacterium lactis, 0.9 part of micrococcus klebsiella, 2.2 parts of bacillus subtilis, 1.0 part of lactobacillus pentosus, 0.8 part of lactobacillus salivarius, 2.5 parts of bacillus cereus, 0.6 part of yeast and 7 parts of protective agent.
Further, the probiotic composition also contains maltose, vitamin A, vitamin C, vitamin D and vitamin B6And microcrystalline cellulose.
Further, the protective agent comprises glycyrrhizic acid and pullulan.
Furthermore, the mass ratio of the glycyrrhizic acid to the pullulan is 1: (2.5-4).
According to the invention, probiotics are reasonably matched, and the ratio of the probiotics is adjusted, so that the effect of the probiotics is exerted to the maximum extent, the intestinal digestion and absorption are promoted, the proliferation of harmful bacteria is inhibited, and the intestinal flora is effectively adjusted; the inventor unexpectedly finds that the glycyrrhizic acid and the pullulan are added into the probiotic composition according to a certain proportion, so that the growth of the probiotics can be enhanced after the probiotics reach the intestinal tract, the activity of the probiotics is effectively improved, and the growth of escherichia coli is inhibited, wherein the glycyrrhizic acid and the pullulan are mixed according to a mass ratio of 1: (2.5-4) when the probiotic composition is added, the activity of the probiotics is better, the intestinal digestion and absorption are better promoted, and the intestinal flora is effectively adjusted.
The second purpose of the invention is to provide a probiotic preparation prepared from the probiotic composition and a medically acceptable carrier, the probiotic preparation has high live bacteria survival rate and can be stored for a long time, the storage time of the probiotic preparation at normal temperature is longer, and the shelf value is improved.
The invention also provides a preparation method of the probiotic preparation, which comprises the following steps:
s1, respectively inoculating lactobacillus bulgaricus, bifidobacterium lactis, micrococcus krusei, bacillus subtilis, lactobacillus pentosus, lactobacillus salivarius, bacillus cereus and saccharomycetes into a fermentation culture medium for fermentation culture, and ending the fermentation culture when the number of viable bacteria reaches a standard to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, and adding microcrystalline cellulose into the fermentation culture medium before fermentation is finished to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, and then adding glycyrrhizic acid, pullulan, maltose, vitamin A, vitamin C, vitamin D and vitamin B6Mixing to obtain compound;
and S4, sequentially granulating, rounding and vacuum drying the compound obtained in the step S3 to obtain the probiotic preparation.
Further, the viable count in the step S1 is 3.5-4.5 × 109And (5) finishing the fermentation culture when cfu/ml is reached.
Furthermore, the main components of the fermentation medium comprise 85-90 g/L of glucose, 5-14 g/L of yeast extract, 10-12 g/L of sodium citrate, 1-2.6 g/L, L-dipotassium phosphate, 0.2-0.6 g/L of cysteine hydrochloride and 2-4 g/L of magnesium sulfate.
The preparation method of the probiotic preparation is simple, safe, pollution-free and low in production cost, and maltose, vitamin A, vitamin C, vitamin D and vitamin B are added into the probiotic preparation6According to the invention, microcrystalline cellulose is added into the fermentation culture medium before the fermentation in step S2 is finished, so that the stability of the probiotic preparation is improved, the storage time of the probiotic preparation is prolonged, and the storage time of the probiotic preparation at normal temperature is prolonged.
In summary, the invention includes at least one of the following beneficial technical effects:
1. the invention provides a probiotic composition, which has high live bacteria survival rate, can promote intestinal digestion and absorption, and effectively regulates the ecological system balance of intestinal flora;
2. the invention provides a probiotic composition, wherein glycyrrhizic acid and pullulan are added into the probiotic composition according to a certain proportion, so that the growth of probiotics can be enhanced after the probiotics reach the intestinal tract, the activity of the probiotics is effectively improved, and the activity of escherichia coli is inhibited;
the invention provides a probiotic preparation, which is characterized in that microcrystalline cellulose is added before the fermentation of streptococcus thermophilus and lactobacillus acidophilus is finished, so that the fermentation time can be reduced, the storage time of the probiotic preparation can be prolonged, and the shape of the probiotic preparation is more stable.
Detailed Description
The present invention will be described in further detail below.
Example 1A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
0.5 part of streptococcus thermophilus, 0.5 part of lactobacillus acidophilus, 0.5 part of lactobacillus bulgaricus, 0.5 part of bifidobacterium lactis, 0.5 part of micrococcus kluyveri, 1.5 parts of bacillus subtilis, 0.5 part of lactobacillus pentosus, 0.5 part of lactobacillus salivarius, 2 parts of bacillus cereus, 0.5 part of yeast and 6 parts of protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 2.5.
Example 2A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
1.5 parts of streptococcus thermophilus, 1 part of lactobacillus acidophilus, 1 part of lactobacillus bulgaricus, 1 part of bifidobacterium lactis, 0.8 part of micrococcus krusei, 1.7 parts of bacillus subtilis, 1 part of lactobacillus pentosus, 1 part of lactobacillus salivarius, 3 parts of bacillus cereus, 0.7 part of yeast and 8 parts of protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1:3.
Example 3A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
2 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.5 parts of bifidobacterium lactis, 1 part of micrococcus keschnei, 2 parts of bacillus subtilis, 1.2 parts of lactobacillus pentosus, 1 part of lactobacillus salivarius, 3 parts of bacillus cereus, 0.8 part of yeast and 9 parts of protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 3.5.
Example 4A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
4 parts of streptococcus thermophilus, 2 parts of lactobacillus acidophilus, 2.5 parts of lactobacillus bulgaricus, 2 parts of bifidobacterium lactis, 1.5 parts of micrococcus krusei, 2.5 parts of bacillus subtilis, 1.5 parts of lactobacillus pentosus, 1.2 parts of lactobacillus salivarius, 4 parts of bacillus cereus, 1 part of yeast and 10 parts of protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 4.
Example 5A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
1.8 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.6 parts of bifidobacterium lactis, 0.9 part of micrococcus klebsiella, 2.2 parts of bacillus subtilis, 1.0 part of lactobacillus pentosus, 0.8 part of lactobacillus salivarius, 2.5 parts of bacillus cereus, 0.6 part of yeast and 7 parts of protective agent.
The protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 3.2.
Example 6A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
0.5 part of streptococcus thermophilus, 0.5 part of lactobacillus acidophilus, 0.5 part of lactobacillus bulgaricus, 0.5 part of bifidobacterium lactis, 0.5 part of micrococcus kluyveri, 1.5 parts of bacillus subtilis, 0.5 part of lactobacillus pentosus, 0.5 part of lactobacillus salivarius, 2 parts of bacillus cereus, 0.5 part of saccharomycete, 0.6 part of maltose, 0.1 part of vitamin A, 0.3 part of vitamin C, 0.3 part of vitamin D, and vitamin B60.2 part, 2 parts of microcrystalline cellulose and 6 parts of a protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 2.5.
Example 7A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
1.5 parts of streptococcus thermophilus, 1 part of lactobacillus acidophilus, 1 part of lactobacillus bulgaricus, 1 part of bifidobacterium lactis, 0.8 part of micrococcus krusei, 1.7 parts of bacillus subtilis, 1 part of lactobacillus pentosus, 1 part of lactobacillus salivarius, 3 parts of bacillus cereus, 0.7 part of saccharomycete, 0.5 part of maltose, 0.2 part of vitamin A, 0.2 part of vitamin C, 0.4 part of vitamin D, and vitamin B60.2 part, 1 part of microcrystalline cellulose and 8 parts of a protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1:3.
Example 8A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
2 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.5 parts of bifidobacterium lactis, 1 part of micrococcus klebsiella, 2 parts of bacillus subtilis, 1.2 parts of lactobacillus pentosus, 1 part of lactobacillus salivarius, 3 parts of bacillus cereus, 0.8 part of saccharomycete, 0.6 part of maltose, 0.1 part of vitamin A, 0.1 part of vitamin C, 0.4 part of vitamin D, and vitamin B60.4 part, 1.8 parts of microcrystalline cellulose and 9 parts of a protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 3.5.
Example 9A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
4 parts of streptococcus thermophilus, 2 parts of lactobacillus acidophilus, 2.5 parts of lactobacillus bulgaricus, 2 parts of bifidobacterium lactis, 1.5 parts of micrococcus krusei, 2.5 parts of bacillus subtilis, 1.5 parts of lactobacillus pentosus, 1.2 parts of lactobacillus salivarius, 4 parts of bacillus cereus, 1 part of saccharomycete, 0.5 part of maltose, 0.3 part of vitamin A, 0.3 part of vitamin C, 0.2 part of vitamin D, and vitamin B60.2 part, 2 parts of microcrystalline cellulose and 10 parts of a protective agent;
the protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 4.
Example 10A probiotic composition
The probiotic composition comprises the following raw materials in parts by weight:
1.8 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.6 parts of bifidobacterium lactis, 0.9 part of micrococcus klebsiella, 2.2 parts of bacillus subtilis, 1.0 part of lactobacillus pentosus, 0.8 part of lactobacillus salivarius, 2.5 parts of bacillus cereus, 0.6 part of saccharomycete, 0.6 part of maltose, 0.3 part of vitamin A, 0.3 part of vitamin C, 0.2 part of vitamin D, and vitamin B60.2 part, 1.4 parts of microcrystalline cellulose and 7 parts of a protective agent.
The protective agent comprises glycyrrhizic acid and pullulan, and the mass ratio of the glycyrrhizic acid to the pullulan is 1: 3.2.
Example 11A probiotic preparation and method of preparation
A preparation method of a probiotic preparation comprises the following steps:
s1, respectively inoculating the lactobacillus bulgaricus, the bifidobacterium lactis, the micrococcus krusei, the bacillus subtilis, the lactobacillus pentosus, the lactobacillus salivarius, the bacillus cereus and the yeast in the weight parts of the example 6 into a fermentation culture medium for fermentation culture, wherein the culture temperature is 35 ℃, and the viable count reaches 3.5 × 109When cfu/ml is reached, ending the fermentation culture to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, wherein the temperature of a fermentation culture sample is 35 ℃, and when the number of viable bacteria reaches 4.2 × 109When cfu/ml is needed, adding microcrystalline cellulose in the weight part of the example 6 into a fermentation medium, and fermenting for 1h to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, mixing for 30min, and then adding the glycyrrhizic acid, the pullulan, the maltose, the vitamin A, the vitamin C, the vitamin D and the vitamin B in the weight parts of the embodiment 66Mixing to obtain compound;
s4, sequentially passing the compound obtained in the step S3 through a granulator, a spheronizer and a dryer to obtain a probiotic preparation;
the main components of the fermentation medium comprise 85g/L glucose, 5g/L yeast extract, 10g/L sodium citrate, 1g/L, L-dipotassium phosphate-0.2 g/L cysteine hydrochloride and 2g/L magnesium sulfate.
Example 12A probiotic formulation and method of preparation
A preparation method of a probiotic preparation comprises the following steps:
s1, respectively inoculating the lactobacillus bulgaricus, the bifidobacterium lactis, the micrococcus krusei, the bacillus subtilis, the lactobacillus pentosus, the lactobacillus salivarius, the bacillus cereus and the yeast in the weight parts of the example 7 into a fermentation culture medium for fermentation culture, wherein the culture temperature is 30 ℃, and the viable count reaches 3.8 × 109When cfu/ml is reached, ending the fermentation culture to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, wherein the temperature of a fermentation culture sample is 35 ℃, and when the number of viable bacteria reaches 4.0 × 109When cfu/ml is needed, adding microcrystalline cellulose in the weight part of the example 7 into a fermentation medium, and fermenting for 1.5h to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, mixing for 35min, and then adding the glycyrrhizic acid, the pullulan, the maltose, the vitamin A, the vitamin C, the vitamin D and the vitamin B in the weight parts of the embodiment 76Mixing to obtain compound;
s4, sequentially granulating, rounding, drying at low temperature in vacuum and coating the compound obtained in the step S3 by using a granulator, a rounding machine, a dryer and a coating machine to obtain the probiotic preparation;
the main components of the fermentation medium comprise 87g/L glucose, 10g/L yeast extract, 11g/L sodium citrate, 1.5g/L, L-dipotassium phosphate-0.4 g/L cysteine hydrochloride and 3g/L magnesium sulfate.
Example 13A probiotic preparation and method of preparation
A preparation method of a probiotic preparation comprises the following steps:
s1, respectively taking the weight portions of the product obtained in the example 8Inoculating Lactobacillus gasseri, Bifidobacterium lactis, Micrococcus clarkii, Bacillus subtilis, Lactobacillus pentosus, Lactobacillus salivarius, Bacillus cereus and yeast into fermentation medium for fermentation culture at 40 deg.C until viable count reaches 4.2 × 109When cfu/ml is reached, ending the fermentation culture to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, wherein the temperature of a fermentation culture sample is 30 ℃, and when the number of viable bacteria reaches 4.5 × 109When cfu/ml is required, adding microcrystalline cellulose in the weight part of the example 8 into a fermentation medium, and fermenting for 50min to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, mixing for 40min, and then adding the glycyrrhizic acid, the pullulan, the maltose, the vitamin A, the vitamin C, the vitamin D and the vitamin B in the weight parts of the embodiment 86Mixing to obtain compound;
s4, sequentially granulating, rounding, drying at low temperature in vacuum and coating the compound obtained in the step S3 by using a granulator, a rounding machine, a dryer and a coating machine to obtain the probiotic preparation;
the main components of the fermentation medium comprise 88g/L of glucose, 12g/L of yeast extract, 11g/L of sodium citrate, 2.3g/L, L g/L of dipotassium phosphate-0.5 g/L of cysteine hydrochloride and 3.5g/L of magnesium sulfate.
Example 14A probiotic preparation and method of preparation
A preparation method of a probiotic preparation comprises the following steps:
s1, respectively inoculating the lactobacillus bulgaricus, the bifidobacterium lactis, the micrococcus krusei, the bacillus subtilis, the lactobacillus pentosus, the lactobacillus salivarius, the bacillus cereus and the yeast in the weight parts of the example 9 into a fermentation culture medium for fermentation culture, wherein the culture temperature is 40 ℃, and the viable count reaches 4.5 × 109When cfu/ml is reached, ending the fermentation culture to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, wherein the temperature of a fermentation culture sample is 35 ℃, and when the number of viable bacteria isUp to 3.8 × 109When cfu/ml is required, adding microcrystalline cellulose in the weight part of the example 9 into a fermentation medium, and fermenting for 1h to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, mixing for 40min, and then adding the glycyrrhizic acid, the pullulan, the maltose, the vitamin A, the vitamin C, the vitamin D and the vitamin B in the weight parts of the example 96Mixing to obtain compound;
s4, sequentially granulating, rounding, drying at low temperature in vacuum and coating the compound obtained in the step S3 by using a granulator, a rounding machine, a dryer and a coating machine to obtain the probiotic preparation;
the main components of the fermentation medium comprise 90g/L glucose, 14g/L yeast extract, 12g/L sodium citrate, 2.6g/L, L-dipotassium phosphate and 0.6g/L cysteine hydrochloride and 4g/L magnesium sulfate.
Example 15A probiotic formulation and method of preparation
A preparation method of a probiotic preparation comprises the following steps:
s1, respectively inoculating the lactobacillus bulgaricus, the bifidobacterium lactis, the micrococcus krusei, the bacillus subtilis, the lactobacillus pentosus, the lactobacillus salivarius, the bacillus cereus and the yeast in the weight parts of the example 10 into a fermentation culture medium for fermentation culture, wherein the culture temperature is 40 ℃, and the viable count reaches 4.3 × 109When cfu/ml is reached, ending the fermentation culture to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, wherein the temperature of a fermentation culture sample is 35 ℃, and when the number of viable bacteria reaches 3.5 × 109When cfu/ml is required, adding microcrystalline cellulose in the weight part of example 10 into a fermentation medium, and fermenting for 1h to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, mixing for 40min, and then adding the glycyrrhizic acid, the pullulan, the maltose, the vitamin A, the vitamin C, the vitamin D and the vitamin B in the weight parts of the embodiment 106Mixing to obtain compound;
s4, sequentially granulating, rounding, drying at low temperature in vacuum and coating the compound obtained in the step S3 by using a granulator, a rounding machine, a dryer and a coating machine to obtain the probiotic preparation;
the main components of the fermentation medium comprise 85-90 g/L of glucose, 5-14 g/L of yeast extract, 10-12 g/L of sodium citrate, 1-2.6 g/L, L-dipotassium phosphate, 0.2-0.6 g/L of cysteine hydrochloride and 2-4 g/L of magnesium sulfate.
Comparative example 1A probiotic preparation
Similar to example 15, except that: the probiotic preparation was free of glycyrrhizic acid and pullulan, and the remaining parameters were the same as in example 5.
Comparative example 2 a probiotic preparation
Similar to example 15, except that: the protective agent is added with glycyrrhizic acid only, and the other parameters are the same as the embodiment 15.
Comparative example 3A probiotic preparation
Similar to example 15, except that: the protective agent only contains pullulan, and the rest parameters are the same as those in example 15.
Comparative example 4 a probiotic preparation
Similar to example 15, except that: the mass ratio of glycyrrhizic acid to pullulan is 1: 1, the remaining parameters are the same as in example 15.
Comparative example 5A probiotic preparation
Similar to example 15, except that: the probiotic formulation was without addition of microcrystalline cellulose and the remaining parameters were the same as in example 15.
Comparative example 6A probiotic preparation
Similar to example 15, except that: microcrystalline cellulose was added after the fermentation of S.thermophilus and L.acidophilus was completed, and the remaining parameters were the same as in example 15.
Test example I determination of viable cell count
The probiotic preparations prepared in examples 11 to 15 and comparative examples 1 to 5 were stored in an environment with a temperature of 37 ± 2 ℃ and a humidity of 75 ± 5%, and the number of viable bacteria in the preparations was counted on days 0, 30 and 60, and the survival rate of the probiotic bacteria was calculated as the number of viable bacteria at the end of day 30 or at the end of day 60/the number of viable bacteria at day 0 × 100%, and the results are shown in table 1.
TABLE 1
Group of Viable count/(cfu/g) at day 0 1 month survival/%) Survival at month 2/%)
Example 11 8.9×109 87 82
Example 12 9.5×109 85 81
Example 13 9.2×109 91 84
Example 14 9.4×109 93 87
Example 15 10.1×109 98 95
Comparative example 1 4.4×109 56 42
Comparative example 2 5.9×109 63 53
Comparative example 3 6.3×109 71 64
Comparative example 4 8.4×109 82 73
Comparative example 5 8.8×109 86 80
According to the data in table 1, the probiotic preparations prepared in examples 11 to 15 have a high viable bacteria count, and after 30 days and 60 days, the viable bacteria survival rate in the probiotic is still high, so that intestinal digestion and absorption can be promoted, and the balance of the intestinal flora ecosystem can be effectively adjusted, the probiotic preparation prepared in comparative example 1 (without adding a protective agent) has a lower viable bacteria count than the probiotic preparations prepared in examples 11 to 15, and the viable bacteria survival rate is lower, the protective agent in the probiotic preparation prepared in comparative example 2 or comparative example 3 only contains one of glycyrrhizic acid or pullulan, the viable bacteria count is lower, the survival rate is lower, the combination of glycyrrhizic acid and pullulan can improve the activity of the probiotic, compared with the probiotic preparation prepared in example 15, the viable bacteria count in comparative example 4 and the survival rate of bacteria are lower than those in example 15, the glycyrrhizic acid and the pullulan are required to be compounded according to a specific proportion so as to better improve the number of viable bacteria and the survival rate of bacteria; the probiotic preparation prepared in comparative example 5 (without microcrystalline cellulose addition) had little effect on the number of viable bacteria and the survival rate of the bacteria.
Test example two, effect of probiotic formulation on intestinal flora
The materials comprise 120 Kunming mice, half male and female, 22-24 g weight for feeding for 7 days, the Kunming mice are randomly divided into 12 groups, each group comprises 10 mice, the probiotic preparation prepared in examples 11-15 and comparative examples 1-5 is used, physiological saline is blank group, and positive group (10% intestines and stomach solution purchased from Beijing Bailin Kangyuan biotechnology Limited liability company, lactobacillus acidophilus is more than or equal to 7.5 × 107Enterococcus faecalis greater than or equal to 2.5 × 107) Feeding 0.2 mL/mouse in each group, wherein the test period is 4 weeks, collecting blood from eyeball of mouse at 4 weeks, aseptically picking thymus and spleen, collecting caecum content, and placing in 10% neutral formaldehyde buffer solution;
10% neutral formaldehyde buffer solution: accurately weighing 0.4gKH2PO4、0.65gK2HPO4Dissolved in a small amount of distilled water, stirred to mix well, and then 10mL of 40% formaldehyde solution was added thereto to adjust pH to 10 mL.
Mixing the collected cecum content and normal saline uniformly at a ratio of 1:9, placing on an oscillator, shaking for 20min, diluting by 10 times, uniformly coating on eosin methylene blue culture medium and MRS culture medium, respectively measuring the number of Escherichia coli and lactobacillus in cecum, placing in an aerobic incubator and an anaerobic incubator at 37 ℃ for 24h and 36h, respectively, and calculating the number of two types of bacteria in each gram of cecum content according to counting results.
TABLE 2
Figure BDA0002509732370000101
Figure BDA0002509732370000111
According to the data in table 2, after week 2, the difference between the colony numbers of the lactic acid bacteria in examples 11 to 15 and the colony numbers of the lactic acid bacteria in the blank group is not significant, but the colony numbers of the lactic acid bacteria in the examples 11 to 15 are significantly lower than that in the blank group, and with the increase of the feeding days, after week 4, the colony numbers of the lactic acid bacteria and the colony numbers of the lactic acid bacteria in examples 11 to 15 are significantly higher and lower than that in the blank group, respectively, and the colony numbers of the lactic acid bacteria in examples 11 to 15 are significantly increased (P < 0.01), and the colony numbers of the lactic acid bacteria are significantly reduced, which indicates that the probiotic preparation of the invention can promote the proliferation of the lactic acid bacteria, inhibit the propagation of the lactic acid bacteria, effectively regulate the balance of the.
Compared with the embodiment 15, the comparative example 1 has the advantages that the protective agent is not added, the number of the lactobacillus colonies is changed, the lactobacillus is easily affected and inactivated by bile salt in the intestinal tract, the number of the lactobacillus colonies is reduced, the number of the escherichia coli colonies is higher, the addition of the protective agent can enhance the number of the lactobacillus colonies, the number of the escherichia coli colonies is reduced, and the digestion and absorption are facilitated.
Compared with the example 15, the comparative example 2 or the comparative example 3 is only added with one of glycyrrhizic acid and pullulan, the increase of the colony number of the lactic acid bacteria is not large, the inhibition effect on the colony number of the escherichia coli is not strong, and the combination of the glycyrrhizic acid and the pullulan can improve the colony number of the lactic acid bacteria and inhibit the colony number of the escherichia coli.
Compared with the probiotic preparation prepared in the example 15, the probiotic preparation prepared in the example 4 has the effects of promoting the increase of the colony number of the lactic acid bacteria and inhibiting the colony of escherichia coli by changing the ratio of the glycyrrhizic acid to the pullulan.
Compared with the example 15, the probiotic preparation does not contain microcrystalline cellulose, and the effects on promoting the increase of the colony number of the lactic acid bacteria and inhibiting the colony number of escherichia coli are similar to and less than the effects of the example 15.
Test example three, accelerated test
The probiotic preparations prepared in examples 11 to 15 and comparative examples 1 to 6 were stored for 6 months at a temperature of 40 ℃. + -. 2 ℃ and a relative humidity of 75%. + -. 5%. Samples were taken at the end of 1 month, 2 months, 3 months, 6 months during the test period and the shape of the probiotic preparation was observed.
The results show that the probiotic preparations of examples 11 to 15 and comparative examples 1 to 4 of the present invention can be stored for a longer period of time without significant changes in the shape of the preparations, which indicates that the addition of microcrystalline cellulose before the fermentation of streptococcus thermophilus and lactobacillus acidophilus can increase the storage time of the probiotic preparations at room temperature, and thus the shape of the probiotic preparations is more stable. In contrast, in comparative example 5, no microcrystalline cellulose was added, and the formulation shape of the probiotic bacteria was dissolved, and in comparative example 6, the microcrystalline cellulose was dissolved after the fermentation of streptococcus thermophilus and lactobacillus acidophilus, and therefore, it was demonstrated that the shape of the formulation could be maintained and the shelf life of the probiotic formulation could be extended only by adding microcrystalline cellulose before the fermentation of streptococcus thermophilus and lactobacillus acidophilus was completed.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims (10)

1. The probiotic composition is characterized by comprising the following raw materials in parts by weight: 0.5-4 parts of streptococcus thermophilus, 0.5-2 parts of lactobacillus acidophilus, 0.5-2.5 parts of lactobacillus bulgaricus, 0.5-2 parts of bifidobacterium lactis, 0.5-1.5 parts of micrococcus krusei, 1.5-2.5 parts of bacillus subtilis, 0.5-1.5 parts of lactobacillus pentosus, 0.5-1.2 parts of lactobacillus salivarius, 2-4 parts of bacillus cereus, 0.5-1 part of saccharomycete and 6-10 parts of protective agent.
2. The probiotic composition according to claim 1, characterized by comprising the following raw materials in parts by weight: 0.5-2 parts of streptococcus thermophilus, 1-2 parts of lactobacillus acidophilus, 1-2.5 parts of lactobacillus bulgaricus, 1-2 parts of bifidobacterium lactis, 0.5-1 part of micrococcus krusei, 2-2.5 parts of bacillus subtilis, 0.5-1.5 parts of lactobacillus pentosus, 0.5-1.0 part of lactobacillus salivarius, 2-3 parts of bacillus cereus, 0.5-1 part of saccharomycete and 6-9 parts of a protective agent.
3. The probiotic composition according to claim 1, characterized by comprising the following raw materials in parts by weight: 1.8 parts of streptococcus thermophilus, 1.5 parts of lactobacillus acidophilus, 2 parts of lactobacillus bulgaricus, 1.6 parts of bifidobacterium lactis, 0.9 part of micrococcus klebsiella, 2.2 parts of bacillus subtilis, 1.0 part of lactobacillus pentosus, 0.8 part of lactobacillus salivarius, 2.5 parts of bacillus cereus, 0.6 part of yeast and 7 parts of protective agent.
4. The probiotic composition according to claim 1, characterized in that it further comprises maltose, vitamin a, vitamin C, vitamin D, vitamin B6And microcrystalline cellulose.
5. The probiotic composition according to claim 1, characterized in that the protective agent comprises glycyrrhizic acid and pullulan.
6. The probiotic composition according to claim 5, wherein the mass ratio of glycyrrhizic acid to pullulan is 1: (2.5-4).
7. A probiotic preparation prepared from the probiotic composition of any one of claims 1 to 6 and a pharmaceutically acceptable carrier.
8. The method for preparing a probiotic preparation according to claim 7, characterized in that it comprises the following steps:
s1, respectively inoculating lactobacillus bulgaricus, bifidobacterium lactis, micrococcus krusei, bacillus subtilis, lactobacillus pentosus, lactobacillus salivarius, bacillus cereus and saccharomycetes into a fermentation culture medium for fermentation culture, and ending the fermentation culture when the number of viable bacteria reaches a standard to obtain a fermentation culture solution I;
s2, inoculating streptococcus thermophilus and lactobacillus acidophilus into a fermentation culture medium for fermentation culture, and adding microcrystalline cellulose into the fermentation culture medium before fermentation is finished to obtain a fermentation culture solution II;
s3, mixing the fermentation culture solution I obtained in the step S1 with the fermentation culture solution II obtained in the step S2, and then adding glycyrrhizic acid, pullulan, maltose, vitamin A, vitamin C, vitamin D and vitamin B6Mixing to obtain compound;
and S4, sequentially granulating, rounding and vacuum drying the compound obtained in the step S3 to obtain the probiotic preparation.
9. The method for preparing probiotic preparation according to claim 8, wherein the number of viable bacteria in step S1 is 3.5-4.5 × 109And (5) finishing the fermentation culture when cfu/ml is reached.
10. The preparation method of the probiotic preparation according to claim 8, wherein the main components of the fermentation medium comprise 85-90 g/L of glucose, 5-14 g/L of yeast extract, 10-12 g/L of sodium citrate, 1-2.6 g/L, L g/L of dipotassium hydrogen phosphate and 0.2-0.6 g/L of cysteine hydrochloride, and 2-4 g/L of magnesium sulfate.
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