CN114045233A - Direct-vat-set fermentation inoculant for relieving constipation-predominant irritable bowel syndrome and preparation method and application thereof - Google Patents

Direct-vat-set fermentation inoculant for relieving constipation-predominant irritable bowel syndrome and preparation method and application thereof Download PDF

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CN114045233A
CN114045233A CN202111293299.0A CN202111293299A CN114045233A CN 114045233 A CN114045233 A CN 114045233A CN 202111293299 A CN202111293299 A CN 202111293299A CN 114045233 A CN114045233 A CN 114045233A
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bowel syndrome
irritable bowel
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relieving constipation
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CN114045233B (en
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方曙光
董瑶
汪欣
朱建国
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Shanghai Yiweierkang Biotechnology Co ltd
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    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus
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    • 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
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis

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Abstract

The invention relates to a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, a preparation method and an application thereof, wherein the direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome comprises a streptococcus thermophilus ST81 fermented product, a lactobacillus bulgaricus LB42 fermented product, a bifidobacterium lactis BLA80 fermented product and a protective agent. The bifidobacterium lactis BLA80 plays a key role in regulating intestinal health and relieving constipation-type irritable bowel syndrome, and is combined with streptococcus thermophilus ST81 and lactobacillus bulgaricus LB42, so that on one hand, the three floras are mutually promoted, the fermentation process is facilitated, the growth of beneficial floras is promoted, on the other hand, the three floras are mutually matched, the remarkable synergistic effect is achieved in regulating intestinal floras and relieving constipation-type irritable bowel syndrome, and the prepared fermented food can improve symptoms such as constipation and abdominal pain, so that defecation is smoother, and the constipation-type irritable bowel syndrome is effectively relieved.

Description

Direct-vat-set fermentation inoculant for relieving constipation-predominant irritable bowel syndrome and preparation method and application thereof
Technical Field
The invention belongs to the technical field of food processing, relates to a direct-vat-set microbial inoculum, a preparation method and application thereof, and particularly relates to a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, a preparation method and application thereof.
Background
Irritable Bowel Syndrome (IBS) is a group of bowel dysfunction diseases with persistent or intermittent episodes, characterized by changes in abdominal pain, abdominal distension, bowel habits and/or stool behavior to clinical manifestations, but a lack of structural and biochemical abnormalities in the gastrointestinal tract. Constipation-predominant irritable bowel syndrome (IBS-C) is one of the more common types of irritable bowel syndrome. The patient may have a series of digestive tract symptoms, such as abdominal pain, abdominal distension, abdominal discomfort, etc., and may also be accompanied by psychological problems such as anxiety, depression, etc. At present, the constipation irritable bowel syndrome is mainly treated by medicaments.
For example, CN112843143A discloses a Chinese medicinal composition for treating constipation-predominant irritable bowel syndrome, and a medicament and an application thereof, wherein the composition of the raw materials is as follows: cortex Magnolia officinalis, flos Magnoliae officinalis, fructus Aurantii Immaturus, fructus Hippophae, thallus laminariae, fructus Cannabis, Atractylodis rhizoma, radix Angelicae sinensis, fructus Aurantii, Cistanchis herba, and Mori fructus. Adding pharmaceutically conventional adjuvants into the composition according to pharmaceutically conventional process, and making into pharmaceutically common dosage forms. The traditional Chinese medicine composition disclosed by the invention shows a definite curative effect on constipation-type irritable bowel syndrome in both animal experiments and clinical symptoms.
CN109731092A discloses a traditional Chinese medicine composition for treating constipation-predominant irritable bowel syndrome and a preparation method thereof. The composition comprises the following raw materials: radix bupleuri, immature bitter orange, white paeony root, fructus cannabis, betel nut, coptis chinensis, garden euphorbia herb, fiveleaf akebia fruit, cluster mallow fruit, dried ginger, poria cocos, momordica grosvenori and liquorice. The medicines are combined according to a specific weight proportion, can treat both liver and spleen, tonify and treat both symptoms and root causes, achieve the effects of dissolving turbidity and detoxifying, clearing hollow viscera and discharging turbidity, can effectively improve symptoms of constipation, abdominal pain and the like of IBS-C, has definite curative effect, high cure rate, low recurrence rate and low price, and can reduce the treatment cost of IBS-C.
However, the drug treatment often causes certain dependence, constipation recurs once a patient stops using the drug, and the drug treatment may cause side effects such as headache, abdominal pain, diarrhea, cardiovascular diseases and the like.
Therefore, how to develop a new method for non-drug treatment of constipation-predominant irritable bowel syndrome is a problem to be urgently solved by those skilled in the art.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a direct vat set, a preparation method and application thereof, and particularly provides a direct vat set for relieving constipation-predominant irritable bowel syndrome, a preparation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a direct-administration type fermentation inoculant for relieving constipation-predominant irritable bowel syndrome, which comprises a streptococcus thermophilus ST81 ferment, a lactobacillus bulgaricus LB42 ferment, a bifidobacterium lactis BLA80 ferment and a protective agent.
The invention creatively combines the fermentation product of streptococcus thermophilus ST81, the fermentation product of lactobacillus bulgaricus LB42 and the fermentation product of bifidobacterium lactis BLA80 to form a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome. The bifidobacterium lactis belongs to probiotics, can play a role in regulating intestinal flora by directly supplementing probiotics in vivo, and can inhibit the growth of harmful flora, so that the intestinal tract maintains a normal microenvironment and is beneficial to the health of the intestinal tract. It has certain regulating effect on functional intestinal diseases including irritable bowel syndrome, intestinal dysfunction, chronic diarrhea, abdominal distention and even chronic constipation. Streptococcus thermophilus is a lactic acid bacterium, has the effects of improving gastrointestinal tract microbial environment, reducing intestinal tract pH value, promoting enterokinesia, preventing colonization of pathogenic bacteria, secreting bacteriocin to inhibit growth of pathogenic bacteria, inhibiting activity of cholesterol synthase, and reducing cholesterol content in blood serum. The fermentation product can regulate and control blood pressure, the products such as polysaccharide, bacteriocin, lactic acid and the like generated by the fermentation have the function of anti-tumor activity, the generation of tumors is resisted by activating an immune system of an organism and inhibiting the mutation of cells, and the superoxide dismutase generated by the fermentation can eliminate excessive superoxide anion free radicals generated in the in-vivo metabolic process and delay aging. The lactobacillus bulgaricus also belongs to lactic acid bacteria, has the effects of helping lactose fermentation and promoting digestion and absorption, can inhibit the growth and development of harmful flora in intestinal tracts, has the effect of regulating the intestinal flora, can activate the immune function in the intestinal tracts, inhibits cancer cell proliferation, and has certain anti-inflammatory and antibacterial effects. In the direct-vat-set fermentation inoculant for relieving constipation-predominant irritable bowel syndrome provided by the invention, bifidobacterium lactis BLA80 plays a key role in regulating intestinal health and relieving constipation-predominant irritable bowel syndrome, and is combined with streptococcus thermophilus ST81 and lactobacillus bulgaricus LB42, so that on one hand, three floras are mutually promoted, the fermentation process is facilitated, the growth of beneficial floras is promoted, on the other hand, the three floras are mutually matched, a remarkable synergistic effect is achieved in regulating intestinal floras and relieving constipation-predominant irritable bowel syndrome, and the prepared fermented food can improve symptoms such as constipation and abdominal pain, so that defecation is smoother, and the constipation-predominant irritable bowel syndrome is effectively relieved.
Preferably, the direct vat set for relieving constipation irritable bowel syndrome comprises 25-45 parts of streptococcus thermophilus ST81 leavening, 1.5-2.5 parts of lactobacillus bulgaricus LB42 leavening, 2.5-65 parts of bifidobacterium lactis BLA80 leavening and 18-36 parts of a protective agent in parts by weight.
Specific values in the above-mentioned 25 to 45 parts may be 25 parts, 27 parts, 29 parts, 31 parts, 33 parts, 35 parts, 37 parts, 39 parts, 41 parts, 43 parts, 45 parts and the like.
Specific values in the above 1.5 to 2.5 parts may be 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts, 2.5 parts and the like.
Specific numerical values in the above 2.5 to 65 parts may be 2.5 parts, 3 parts, 3.5 parts, 4 parts, 8 parts, 12 parts, 16 parts, 20 parts, 24 parts, 28 parts, 32 parts, 36 parts, 40 parts, 44 parts, 48 parts, 52 parts, 56 parts, 60 parts, 62 parts, 65 parts and the like.
Specific numerical values among the above 18 to 36 parts may be 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts and the like.
Preferably, the number of viable bacteria of bifidobacterium lactis BLA80 in the direct vat set for relieving constipation-predominant irritable bowel syndrome is 1 × 1011cfu/g or more, e.g. 1X 1011cfu/g、2×1011cfu/g、3×1011cfu/g、 4×1011cfu/g、6×1011cfu/g、8×1011cfu/g、9×1011cfu/g、1×1012cfu/g, and the like.
Preferably, the protective agent comprises any one or a combination of at least two of trehalose, maltodextrin, beta-cyclodextrin, sucrose, glycerol, mannitol or tween-80, the combination of at least two of trehalose and maltodextrin, the combination of beta-cyclodextrin and sucrose, the combination of sucrose and glycerol and the like, any combination mode can be selected, and the description is omitted, and the combination of trehalose, maltodextrin, beta-cyclodextrin, sucrose, glycerol, mannitol and tween-80 is preferred.
Compared with other choices, the combination of trehalose, malt paste, beta-cyclodextrin, sucrose, glycerol, mannitol and tween-80 is adopted as a protective agent, so that the stability of the prepared fermented food is improved.
Preferably, the protective agent comprises 12-18 parts of trehalose, 2.5-8.5 parts of maltodextrin, 1.0-1.5 parts of beta-cyclodextrin, 2-5 parts of sucrose, 1.2-2.4 parts of glycerol, 0.01-0.3 part of mannitol and 0.25 part of tween-800.02 in parts by weight.
The specific numerical value of the above-mentioned 12 to 18 parts may be 12 parts, 12.5 parts, 13 parts, 13.5 parts, 14 parts, 14.5 parts, 15 parts, 15.5 parts, 16 parts, 16.5 parts, 17 parts, 17.5 parts, 18 parts, etc.
Specific numerical values among the above 2.5 to 8.5 parts may be 2.5 parts, 2.7 parts, 3 parts, 3.2 parts, 3.5 parts, 3.7 parts, 4 parts, 4.2 parts, 4.5 parts, 4.7 parts, 5 parts, 5.2 parts, 5.5 parts, 5.7 parts, 6 parts, 6.2 parts, 6.5 parts, 6.7 parts, 7 parts, 7.2 parts, 7.5 parts, 7.7 parts, 8 parts, 8.2 parts, 8.5 parts and the like.
Specific values in the above 1.0 to 1.5 parts may be 1 part, 1.15 parts, 1.2 parts, 1.25 parts, 1.3 parts, 1.35 parts, 1.4 parts, 1.45 parts, 1.5 parts and the like.
Specific values in the above 2 to 5 parts may be 2 parts, 2.2 parts, 2.5 parts, 2.7 parts, 3 parts, 3.2 parts, 3.5 parts, 3.7 parts, 4 parts, 4.2 parts, 4.5 parts, 4.7 parts, 5 parts and the like.
Specific values in the above 1.2 to 2.4 parts may be 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts and the like.
Specific values of the above 0.01 to 0.3 part may be 0.01 part, 0.03 part, 0.05 part, 0.07 part, 0.1 part, 0.12 part, 0.15 part, 0.17 part, 0.2 part, 0.22 part, 0.25 part, 0.27 part, 0.3 part and the like.
Specific values of the above-mentioned 0.02 to 0.25 part may be 0.02 part, 0.04 part, 0.06 part, 0.08 part, 0.1 part, 0.12 part, 0.15 part, 0.17 part, 0.2 part, 0.22 part, 0.25 part and the like.
In a second aspect, the present invention provides a method for preparing a direct vat set for relieving constipation-predominant irritable bowel syndrome according to the first aspect, the method comprising the steps of:
(1) culturing and activating streptococcus thermophilus ST81, and fermenting to obtain streptococcus thermophilus ST81 fermentation product; culturing and activating lactobacillus bulgaricus LB42, and fermenting to obtain lactobacillus bulgaricus LB42 fermentation product; culturing and activating Bifidobacterium lactis BLA80, and fermenting to obtain Bifidobacterium lactis BLA80 fermented product;
(2) and (2) mixing the streptococcus thermophilus ST81 fermentation product, the lactobacillus bulgaricus LB42 fermentation product and the bifidobacterium lactis BLA80 fermentation product obtained in the step (1) with a protective agent to obtain the direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome.
Preferably, the temperature of the culture of Streptococcus thermophilus ST81, the temperature of the culture of Lactobacillus bulgaricus LB42 and the temperature of the culture of Bifidobacterium lactis BLA80 in step (1) are each independently 35 to 40 ℃, for example 35 ℃, 35.5 ℃, 36 ℃, 36.5 ℃, 37 ℃, 37.5 ℃, 38 ℃, 38.5 ℃, 39 ℃, 39.5 ℃, 40 ℃ and the like.
Preferably, the time for culturing the streptococcus thermophilus ST81, the time for culturing the lactobacillus bulgaricus LB42 and the time for culturing the bifidobacterium lactis BLa80 in the step (1) are respectively and independently 16-24h, such as 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h and the like.
Preferably, the temperature of fermentation of Streptococcus thermophilus ST81, the temperature of fermentation of Lactobacillus bulgaricus LB42 and the temperature of fermentation of Bifidobacterium lactis BLA80 in step (1) are each independently 35 to 40 ℃, such as 35 ℃, 35.5 ℃, 36 ℃, 36.5 ℃, 37 ℃, 37.5 ℃, 38 ℃, 38.5 ℃, 39 ℃, 39.5 ℃, 40 ℃ and the like.
Preferably, the time for fermentation of Streptococcus thermophilus ST81, the time for fermentation of Lactobacillus bulgaricus LB42 and the time for fermentation of Bifidobacterium lactis BLA80 in step (1) are each independently 9-12h, such as 9h, 9.2h, 9.5h, 9.7h, 10h, 10.2h, 10.5h, 10.7h, 11h, 11.2h, 11.5h, 11.7h, 12h, etc.
Preferably, the pH of the streptococcus thermophilus ST81 fermentation system, the pH of the lactobacillus bulgaricus LB42 fermentation system and the pH of the bifidobacterium lactis BLa80 fermentation system in step (1) are each independently 5.5-6.0, such as 5.5, 5.55, 5.6, 5.65, 5.7, 5.75, 5.8, 5.85, 5.9, 5.95, 6, etc.
Preferably, the fermentation product is collected by centrifugation after the fermentation of Streptococcus thermophilus ST81, the fermentation of Lactobacillus bulgaricus LB42 and the fermentation of Bifidobacterium lactis BLA80 in step (1).
Preferably, the rotational speeds of the centrifugation are 6000-10000r/min, such as 6000r/min, 6500r/min, 7000r/min, 7500r/min, 8000r/min, 8500r/min, 9000r/min, 9500 r/min, 10000r/min, and the like, independently of one another.
The centrifugation time is 10-15min, such as 10min, 10.5min, 11min, 11.5min, 12min, 12.5min, 13min, 13.5min, 14min, 14.5min, 15min, etc.
Preferably, the mixing in step (2) further comprises drying.
Preferably, the drying is by freeze drying.
In a third aspect, the present invention provides a use of the directly administered fermentation broth for relieving constipation-predominant irritable bowel syndrome according to the first aspect in preparing a fermented food, wherein the inoculation amount of the directly administered fermentation broth for relieving constipation-predominant irritable bowel syndrome is 20-50g/T, such as 20g/T, 22g/T, 25g/T, 27g/T, 30g/T, 32g/T, 35g/T, 37g/T, 40g/T, 42g/T, 45g/T, 47g/T, 50g/T, etc. The T represents ton.
Preferably, the fermented food includes any one of fermented soybean milk, fermented soybean powder, fermented milk, or fermented milk powder.
In a fourth aspect, the present invention provides a fermented soybean milk for relieving constipation-predominant irritable bowel syndrome, which comprises soybean flour, a gum base, water and the direct vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome according to the first aspect.
Preferably, the preparation raw materials of the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome comprise, by mass, 0.002-0.005% of a direct-vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, 10-28% of soybean flour, 3-20% of a gum base material, and the balance of water.
Specific values of the above-mentioned 0.002 to 0.005% may be 0.002%, 0.0022%, 0.0025%, 0.0027%, 0.003%, 0.0032%, 0.0035%, 0.0037%, 0.004%, 0.0042%, 0.0045%, 0.0047%, 0.005%, etc.
Specific values of 10 to 28% may be 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, etc.
Specific values of the above-mentioned 3-20% may be 3%, 4%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, etc.
Preferably, the gum base material comprises any one or combination of at least two of coconut pulp, white granulated sugar, agar, pectin, citrus fiber and sea salt, the combination of at least two of the coconut pulp and the white granulated sugar, the combination of the agar and the pectin, the combination of the pectin and the citrus fiber and the like can be selected, and any combination mode can be selected, so that the detailed description is omitted, and the combination of the coconut pulp, the white granulated sugar, the agar, the pectin, the citrus fiber and the sea salt is preferred.
Preferably, the gum base material comprises, by weight, 3-10 parts of coconut milk, 1-5 parts of white granulated sugar, 0.1-1 part of agar, 0.1-0.8 part of pectin, 0.2-0.8 part of citrus fiber and 0.01-0.1 part of sea salt.
Specific numerical values in the above-mentioned 3 to 10 parts may be 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts, 8.5 parts, 9 parts, 9.5 parts, 10 parts and the like.
Specific values in the above 1 to 5 parts may be 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts and the like.
Specific values in the above 0.1 to 1 part may be 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part, 1 part and the like.
Specific values of the above 0.1 to 0.8 parts may be 0.1 part, 0.15 part, 0.2 part, 0.25 part, 0.3 part, 0.35 part, 0.4 part, 0.45 part, 0.5 part, 0.55 part, 0.6 part, 0.65 part, 0.7 part, 0.75 part, 0.8 part and the like.
Specific values of the above 0.2 to 0.8 parts may be 0.2 parts, 0.25 parts, 0.3 parts, 0.35 parts, 0.4 parts, 0.45 parts, 0.5 parts, 0.55 parts, 0.6 parts, 0.65 parts, 0.7 parts, 0.75 parts, 0.8 parts and the like.
Specific numerical values of the above 0.01 to 0.1 part may be 0.01 part, 0.015 part, 0.02 part, 0.025 part, 0.03 part, 0.035 part, 0.04 part, 0.045 part, 0.05 part, 0.055 part, 0.06 part, 0.065 part, 0.07 part, 0.075 part, 0.08 part, 0.085 part, 0.09 part, 0.095 part, 0.1 part or the like.
In a fifth aspect, the invention provides a preparation method of the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome according to the fourth aspect, wherein the preparation method comprises the steps of mixing soybean flour, a gum base material and water, homogenizing, sterilizing, inoculating a direct vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, fermenting, and demulsifying to obtain the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome.
Preferably, the mixing temperature is 60-90 degrees C, such as 60 degrees C, 62 degrees C, 65 degrees C, 67 degrees C, 70 degrees C, 72 degrees C, 75 degrees C, 77 degrees C, 80 degrees C, 82 degrees C, 85 degrees C, 87 degrees C, 90 degrees C.
Preferably, the temperature of the inoculation is 38-50 deg.C, such as 38 deg.C, 39 deg.C, 40 deg.C, 41 deg.C, 42 deg.C, 43 deg.C, 44 deg.C, 45 deg.C, 46 deg.C, 47 deg.C, 48 deg.C, 49 deg.C, 50 deg.C etc.
Preferably, the fermentation temperature is 38-50 deg.C, such as 38 deg.C, 39 deg.C, 40 deg.C, 41 deg.C, 42 deg.C, 43 deg.C, 44 deg.C, 45 deg.C, 46 deg.C, 47 deg.C, 48 deg.C, 49 deg.C, 50 deg.C, etc.
The recitation of numerical ranges herein includes not only the above-recited values, but also any values between any of the above-recited numerical ranges not recited, and for the sake of brevity and clarity, the present invention is not intended to be exhaustive of the specific values encompassed within the range.
Compared with the prior art, the invention has the following beneficial effects:
the invention creatively combines the fermentation product of streptococcus thermophilus ST81, the fermentation product of lactobacillus bulgaricus LB42 and the fermentation product of bifidobacterium lactis BLA80 to form a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome. The bifidobacterium lactis BLA80 plays a key role in regulating intestinal health and relieving constipation-type irritable bowel syndrome, and is combined with streptococcus thermophilus ST81 and lactobacillus bulgaricus LB42, so that on one hand, the three floras are mutually promoted, the fermentation process is facilitated, the growth of beneficial floras is promoted, on the other hand, the three floras are mutually matched, the synergistic effect is remarkable in regulating intestinal floras and relieving constipation-type irritable bowel syndrome, and the prepared fermented food can improve symptoms such as constipation and abdominal pain, so that defecation is smoother, and the constipation-type irritable bowel syndrome is effectively relieved. In addition, the present invention has the effect of improving the stability of the prepared fermented food by using a specific combination of trehalose, maltodextrin, β -cyclodextrin, sucrose, glycerol, mannitol and tween-80 as a protective agent.
Detailed Description
To further illustrate the technical means and effects of the present invention, the following preferred embodiments of the present invention are further described, but the present invention is not limited to the scope of the embodiments.
The Streptococcus thermophilus ST81 related to the following examples, comparative examples, application examples and comparative application examples is preserved in the general microbiological center of China Committee for culture Collection of microorganisms, and is classified and named as Streptococcus thermophilus, the preservation number is CGMCC No.15752, the preservation date is 2018.05.11, and the preservation address is No. 3 of Xilu No.1 of Xinyang district of Beijing City.
The following examples, comparative examples, application examples and comparative application examples relate to Lactobacillus bulgaricus LB42 which was deposited in the general microbiological center of the China Committee for culture Collection of microorganisms and which was classified and named Lactobacillus delbrueckii subsp. bulgaricus, with the deposit number of CGMCC No.15751, the deposit date of 2018.05.11, and the deposit address of No. 3, North Chen West Lu No.1, of the sunward, Beijing.
The Bifidobacterium lactis BLA80 related to the following examples, comparative examples, application examples and comparative application examples is stored in the China general microbiological culture Collection center, and is classified and named as Bifidobacterium animalis subsp.Lactis, the preservation number of the Bifidobacterium animalis is CGMCC NO.15410, the preservation date of the Bifidobacterium animalis is 2018.03.05, and the preservation address of the Bifidobacterium lactis is No. 3 of the North Chen West Lu No.1 of the sunward region of Beijing.
Example 1
The embodiment provides a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, which comprises 30 parts by weight of streptococcus thermophilus ST81 leavening, 1.5 parts by weight of lactobacillus bulgaricus LB42 leavening, 2.5 parts by weight of bifidobacterium lactis BLA80 leavening, 12.5 parts by weight of trehalose, 4 parts by weight of maltodextrin, 1 part by weight of beta-cyclodextrin, 3 parts by weight of sucrose, 1.5 parts by weight of glycerol, 0.05 part by weight of mannitol and 800.2 parts by weight of tween-800.2. Wherein the viable count of Bifidobacterium lactis BLA80 in the direct-administered fermentation agent for relieving constipation-type irritable bowel syndrome is 1 × 1011cfu/g。
The preparation method comprises the following steps:
(1) inoculating Bifidobacterium lactis BLA80 preserved in Glycine max (L.) Gaertn tube into MRS liquid culture medium at a ratio of 2%, standing at 37 deg.C for 22 hr, and activating for 3 generations. Inoculating the activated seeds into a fermentation tank with the inoculation amount of 2%, culturing at the temperature of 37 ℃ for 11h at the pH of 5.5, collecting bacterial liquid, centrifuging for 10min at the speed of 10000r/min, removing supernatant, and collecting bacterial sludge to obtain the fermented product of the bifidobacterium lactis BLA 80.
The streptococcus thermophilus ST81 preserved in the glycerin tube is inoculated in MRS liquid culture medium according to the proportion of 2 percent, and is statically cultured for 20 hours at the temperature of 38 ℃ to be activated for 3 generations. Inoculating the activated seeds into a fermentation tank with the inoculation amount of 3%, culturing at the pH of 6.0 and the temperature of 39 ℃ for 9h, collecting bacterial liquid, centrifuging at 8000r/min for 12min, discarding supernatant, and collecting bacterial sludge to obtain the streptococcus thermophilus ST81 fermentation product.
The lactobacillus bulgaricus LB42 preserved by glycerin tube is inoculated in MRS liquid culture medium according to the proportion of 2 percent, and is statically cultured for 18h at the temperature of 39 ℃ and activated for 3 generations. Inoculating the activated seeds into a fermentation tank according to the inoculation amount of 2.5%, culturing for 10h at the temperature of 38 ℃ and the pH of 5.8, collecting bacterial liquid, centrifuging for 15min at the speed of 6000r/min, removing supernatant, and collecting bacterial sludge to obtain the lactobacillus bulgaricus LB42 ferment.
(2) And (2) uniformly mixing the bifidobacterium lactis BLA80 fermented product, the streptococcus thermophilus ST81 fermented product and the lactobacillus bulgaricus LB42 fermented product obtained in the step (1) with trehalose, maltodextrin, beta-cyclodextrin, sucrose, glycerol, mannitol and tween-80, and carrying out vacuum freeze drying to obtain the direct-vat-set microbial inoculum for relieving constipation type irritable bowel syndrome.
Example 2
The embodiment provides a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, which comprises 25 parts by weight of streptococcus thermophilus ST81 leavening, 2 parts by weight of lactobacillus bulgaricus LB42 leavening, 2.5 parts by weight of bifidobacterium lactis BLA80 leavening, 14 parts by weight of trehalose, 8 parts by weight of maltodextrin, 1.5 parts by weight of beta-cyclodextrin, 2 parts by weight of sucrose, 2.2 parts by weight of glycerol, 800.25 parts by weight of tween-800 and 0.05 part by weight of mannitol.
The preparation method comprises the following steps:
(1) the streptococcus thermophilus ST81 preserved in the glycerin tube is inoculated in MRS liquid culture medium according to the proportion of 2 percent, and is statically cultured for 22h at 37 ℃ for 3 generations. Inoculating the activated seeds into a fermentation tank according to the inoculation amount of 2%, culturing at the temperature of 37 ℃ for 11h at the pH of 5.5, collecting bacterial liquid, centrifuging for 10min at the speed of 10000r/min, removing supernatant, and collecting bacterial sludge to obtain the streptococcus thermophilus ST81 fermented product.
The lactobacillus bulgaricus LB42 preserved by glycerin tube is inoculated in MRS liquid culture medium according to the proportion of 2 percent, and is statically cultured for 20 hours at the temperature of 38 ℃ and activated for 3 generations. Inoculating the activated seeds into a fermentation tank according to the inoculation amount of 3%, culturing for 9h at the temperature of 39 ℃ and the pH of 6.0, collecting bacterial liquid, centrifuging for 12min at 8000r/min, removing supernatant, and collecting bacterial sludge to obtain the lactobacillus bulgaricus LB42 fermented product.
Inoculating Bifidobacterium lactis BLA80 preserved in Glycine max (L.) Gaertn tube into MRS liquid culture medium at a ratio of 2%, standing at 39 deg.C for 18 hr, and activating for 3 generations. Inoculating the activated seeds into a fermentation tank at the inoculation amount of 2.5%, culturing at pH 5.8 and 38 deg.C for 10h, collecting bacterial liquid, centrifuging at 6000r/min for 15min, removing supernatant, and collecting bacterial mud to obtain fermented product of Bifidobacterium lactis BLA 80.
(2) And (2) uniformly mixing the streptococcus thermophilus ST81 fermentation product, the lactobacillus bulgaricus LB42 fermentation product and the bifidobacterium lactis BLA80 fermentation product obtained in the step (1) with trehalose, maltodextrin, beta-cyclodextrin, sucrose, Tween-80, glycerol and mannitol, and carrying out vacuum freeze drying to obtain the direct-vat-set microbial inoculum for relieving constipation type irritable bowel syndrome.
Example 3
The embodiment provides a direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, which comprises 32 parts by weight of streptococcus thermophilus ST81 leavening, 1.5 parts by weight of lactobacillus bulgaricus LB42 leavening, 2.5 parts by weight of bifidobacterium lactis BLA80 leavening, 13 parts by weight of trehalose, 3 parts by weight of maltodextrin, 1.5 parts by weight of beta-cyclodextrin, 2 parts by weight of sucrose, 0.01 part by weight of mannitol, 1 part by weight of glycerol and 800.2 parts by weight of tween-800.2.
The preparation method comprises the following steps:
(1) inoculating Lactobacillus bulgaricus LB42 preserved in Glycine max (L.) Gaertn tube into MRS liquid culture medium at a ratio of 2%, standing at 37 deg.C for 22 hr, and activating for 3 generations. Inoculating the activated seeds into a fermentation tank according to the inoculation amount of 2%, culturing at the temperature of 37 ℃ for 11h at the pH of 5.5, collecting bacterial liquid, centrifuging at 10000r/min for 10min, removing supernatant, and collecting bacterial sludge to obtain the lactobacillus bulgaricus LB42 fermentation product.
Inoculating Bifidobacterium lactis BLA80 preserved in Glycine max (L.) Gaertn tube into MRS liquid culture medium at a ratio of 2%, standing at 38 deg.C for 20 hr, and activating for 3 generations. Inoculating the activated seeds into a fermentation tank with 3% of inoculation amount, culturing at pH 6.0 and 39 deg.C for 9h, collecting bacterial liquid, centrifuging at 8000r/min for 12min, discarding supernatant, and collecting bacterial sludge to obtain fermented product of Bifidobacterium lactis BLA 80.
The streptococcus thermophilus ST81 preserved in the glycerin tube is inoculated in MRS liquid culture medium according to the proportion of 2 percent, and is statically cultured for 18h at the temperature of 39 ℃ to be activated for 3 generations. Inoculating the activated seeds into a fermentation tank according to the inoculation amount of 2.5%, culturing for 10h at the temperature of 38 ℃ and the pH value of 5.8, collecting bacterial liquid, centrifuging for 15min at the speed of 6000r/min, removing supernatant, and collecting bacterial sludge to obtain the streptococcus thermophilus ST81 fermentation product.
(2) And (2) uniformly mixing the lactobacillus bulgaricus LB42 fermented product, the bifidobacterium lactis BLA80 fermented product and the streptococcus thermophilus ST81 fermented product obtained in the step (1) with trehalose, sucrose, mannitol, maltodextrin, beta-cyclodextrin, glycerol and tween-80, and carrying out vacuum freeze drying to obtain the direct-vat-set fermentation inoculum for relieving the constipation type irritable bowel syndrome.
Example 4
This example provides a direct vat set for relieving constipation-predominant irritable bowel syndrome, which differs from example 1 only in that "trehalose 12.5 parts, maltodextrin 4 parts, beta-cyclodextrin 1 part, sucrose 3 parts, glycerin 1.5 parts, mannitol 0.05 parts and tween-800.2 parts" in example 1 are replaced with "trehalose 13 parts, maltodextrin 5.25 parts, sucrose 2 parts, glycerin 2 parts", and the other components and contents are not changed.
The preparation process is as in example 1.
Example 5
This example provides a direct vat set for relieving constipation-predominant irritable bowel syndrome, which differs from example 1 only in that "trehalose 12.5 parts, maltodextrin 4 parts, beta-cyclodextrin 1 part, sucrose 3 parts, glycerin 1.5 parts, mannitol 0.05 part and tween-800.2 parts" in example 1 are replaced with "trehalose 15 parts and maltodextrin 7.25 parts", and the other components and contents are not changed.
The preparation process is as in example 1.
Comparative example 1
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of S.thermophilus ST81 ferment, 1.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "31.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment", and the other components and contents are not changed.
The preparation process is as in example 1.
Comparative example 2
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of S.thermophilus ST81 ferment, 1.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "31.5 parts of S.thermophilus ST81 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment", and the other components and contents are not changed.
The preparation process is as in example 1.
Comparative example 3
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of the Streptococcus thermophilus ST81 ferment, 1.5 parts of the Lactobacillus bulgaricus LB42 ferment, 2.5 parts of the Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "34 parts of the combination of the Streptococcus thermophilus ST81 ferment and the Lactobacillus bulgaricus LB42 ferment", wherein the mass ratio of the Streptococcus thermophilus ST81 ferment to the Lactobacillus bulgaricus LB42 ferment is 20:1, and the other components and the contents are not changed.
The preparation process is as in example 1.
Comparative example 4
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of S.thermophilus ST81 ferment, 1.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "2.5 parts of Bifidobacterium lactis BLA80 ferment", and other components and contents are not changed.
The preparation process is as in example 1.
Comparative example 5
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of S.thermophilus ST81 ferment, 1.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "34 parts of S.thermophilus ST81 ferment", and the other components and contents are not changed.
The preparation process is as in example 1.
Comparative example 6
This comparative example provides a direct vat set inoculant which differs from example 1 only in that "30 parts of S.thermophilus ST81 ferment, 1.5 parts of Lactobacillus bulgaricus LB42 ferment, 2.5 parts of Bifidobacterium lactis BLA80 ferment" of example 1 are replaced by "34 parts of Lactobacillus bulgaricus LB42 ferment", and the other components and contents are not changed.
The preparation process is as in example 1.
Application example 1
The application example provides fermented soybean milk for relieving constipation-predominant irritable bowel syndrome, and the raw materials for preparing the fermented soybean milk comprise, by mass, 0.003% of the direct vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome in example 1, 17.5% of soybean meal, 5.5% of coconut milk, 2% of white granulated sugar, 1% of agar, 0.3% of pectin, 0.2% of citrus fiber, 0.03% of sea salt and the balance of water.
The preparation method comprises the following steps:
(1) dissolving soybean powder in water, shearing at 80 deg.C for 15min, standing for 10min, mixing, and heating to 90 deg.C. Mixing agar, citrus fiber and white sugar at 70 deg.C, shearing for 20min to dissolve, adding dissolved bean powder, coconut milk, pectin and sea salt, stirring, and mixing to obtain mixture.
(2) Homogenizing the mixture obtained in the step (1) at 60 ℃ and 20 MPa.
(3) Heating the homogenized mixture to 95 deg.C, and maintaining the temperature for 5min for sterilization.
(4) Cooling the sterilized mixture to 43 ℃, inoculating a direct-vat-set fermentation microbial inoculum for relieving constipation-type irritable bowel syndrome, fermenting at 44 ℃, stopping when the pH of the system is below 4.5 and the acidity of the system is above 30 DEG T, and demulsifying to obtain the fermented soybean milk for relieving constipation-type irritable bowel syndrome.
Application example 2
The application example provides fermented soybean milk for relieving constipation-predominant irritable bowel syndrome, and the raw materials for preparing the fermented soybean milk comprise, by mass, 0.003% of the direct vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome of example 2, 25% of soybean meal, 3% of coconut milk, 1% of white granulated sugar, 0.5% of agar, 0.1% of pectin, 0.5% of citrus fiber and the balance of water.
The preparation method comprises the following steps:
(1) dissolving soybean powder in water, shearing at 70 deg.C for 15min, standing for 10min, mixing, and heating to 80 deg.C. Mixing agar, citrus fiber and white sugar at 60 deg.C, shearing for 20min to dissolve, adding dissolved bean powder, pectin and coconut milk, stirring, and mixing to obtain mixture.
(2) Homogenizing the mixture obtained in the step (1) at 65 ℃ and 20 MPa.
(3) Heating the homogenized mixture to 90 deg.C, and maintaining the temperature for 10min for sterilization.
(4) Cooling the sterilized mixture to 48 ℃, inoculating a direct-vat-set fermentation microbial inoculum for relieving constipation-type irritable bowel syndrome, fermenting at 47 ℃, stopping when the pH of a system is below 4.5 and the acidity of the system is above 30 DEG T, and demulsifying to obtain the fermented soybean milk for relieving constipation-type irritable bowel syndrome.
Application example 3
The application example provides fermented soybean milk for relieving constipation-predominant irritable bowel syndrome, and the raw materials for preparing the fermented soybean milk comprise, by mass, 0.003% of the direct vat set for relieving constipation-predominant irritable bowel syndrome of example 3, 15% of soybean meal, 9.5% of coconut milk, 5% of white granulated sugar, 0.2% of agar, 0.08% of sea salt, and the balance of water.
The preparation method comprises the following steps:
(1) dissolving soybean powder in water, shearing at 75 deg.C for 15min, standing for 15min, mixing, and heating to 85 deg.C. Mixing agar and white sugar at 65 deg.C, shearing for 15min to dissolve, adding dissolved bean powder, coconut milk and sea salt, stirring, and mixing to obtain mixture.
(2) Homogenizing the mixture obtained in the step (1) at 70 ℃ and 18 MPa.
(3) Heating the homogenized mixture to 90 deg.C, and maintaining the temperature for 10min for sterilization.
(4) Cooling the sterilized mixture to 38 ℃, inoculating a direct-vat-set fermentation microbial inoculum for relieving constipation-type irritable bowel syndrome, fermenting at 39 ℃, stopping when the pH of a system is below 4.5 and the acidity of the system is above 30 DEG T, and demulsifying to obtain the fermented soybean milk for relieving constipation-type irritable bowel syndrome.
Application examples 4 to 5
Application examples 4 to 5 provide two kinds of fermented soybean milks for relieving constipation-predominant irritable bowel syndrome, which are different from application example 1 only in that "0.003% of the straightly administered fermentation broth for relieving constipation-predominant irritable bowel syndrome of example 1" was replaced with 0.003% of the straightly administered fermentation broth for relieving constipation-predominant irritable bowel syndrome of examples 4 to 5, respectively, and the other components and contents were not changed.
The preparation process is as in application example 1.
Comparative application examples 1 to 6
Comparative application examples 1 to 6 provide six kinds of fermented soybean milks, which are different from application example 1 only in that "0.003% of the orthotropic fermented microbial agent for relieving constipation-predominant irritable bowel syndrome of example 1" was replaced with "0.003% of the orthotropic fermented microbial agent of comparative examples 1 to 6", respectively, and the other components and contents were not changed.
The preparation process is as in application example 1.
Test example 1
And (3) stability testing:
the viscosity and the acidity are important indexes for evaluating the mouthfeel of the fermented soybean milk and also important indexes for evaluating the quality stability of the fermented soybean milk. The viable count of the bifidobacterium lactis BLA80 is an important influence factor for the efficacy of fermented soybean milk and the regulation of intestinal health. Therefore, the fermented soybean milks of application examples 1 to 5 were tested for viscosity, acidity, pH value, total number of lactic acid bacteria and viable cell number change of Bifidobacterium lactis BLA80 in shelf life (refrigerated at 10 ℃ C. for 21 days) by the following methods:
the acidity was measured by an acidity meter (using phenolphthalein as an indicator and titrating with 0.101mol/L NaOH standard solution). The pH was measured with a pH meter (METTLER TOLEDO pH meter). The viscosity of the product was measured using a viscometer (NDJ-8S).
The total number of live lactobacillus (namely the total number of live streptococcus thermophilus and lactobacillus bulgaricus) and the total number of live bifidobacterium (namely the number of live bifidobacterium lactis BLA 80) are detected according to the method of the national standard GB 4789.35-2016. The results are shown in Table 1.
TABLE 1
Figure BDA0003335468890000181
Figure BDA0003335468890000191
The results show that: the fermented soybean milk provided by application examples 1-5 has small viscosity change in shelf life and stable texture state. The acidity and pH of the fermented soybean milk change to a smaller extent on the whole, which indicates that the flavor and the taste are more stable. During the cold storage period, the total number of lactobacillus in the fermented soybean milk is reduced, and the viable count of Bifidobacterium lactis BLA80 is stable and reaches 3.0 × 107More than cfu/g, therefore, people suffering from constipation-predominant irritable bowel syndrome can achieve the effects of regulating intestinal health and relieving constipation by drinking 300g of the fermented soybean milk provided in application examples 1-5 every day. Among them, the viscosity change, acidity change, and pH change degree of the fermented soybean milk provided in application examples 4 and 5 were slightly larger than those of application example 1, and the viable count of lactic acid bacteria and bifidobacterium lactis BLa80 was also slightly lower than that of application example 1, which indicates that the use of a specific combination of trehalose, malt paste, β -cyclodextrin, sucrose, glycerin, mannitol, and tween-80 as a protecting agent has an effect of improving the stability of the prepared fermented soybean milk. In summary, the fermented soybean milks provided in application examples 1 to 5 have small changes in viscosity, acidity and pH during refrigeration, and the fermented soybean milks have a high viable cell count of bifidobacterium lactis BLa80 and are stable, indicating that the fermented soybean milks have good stability.
Test example 2
And (3) safety evaluation:
the safety of the fermented soybean milk provided in application examples 1 to 5 was evaluated. 100 healthy volunteers (without allergic history to fermented soybean milk) were selected and randomly divided into 5 groups of 20, and each of the 1 st to 5 th groups took 300ml of fermented soybean milk provided in application examples 1 to 5 per day. All volunteers did not take other drugs for treating gastrointestinal diseases and other fermented foods during the test period. The test period was 1 month. And recording and comparing the occurrence conditions of adverse reactions such as abdominal pain, abdominal distension, nausea and the like of all groups of volunteers, and evaluating the safety of the fermented soybean milk.
The results show that: all volunteers do not have the phenomena of abdominal pain, abdominal distension, nausea and the like, which shows that the fermented soybean milk provided by the application examples 1-5 has no adverse reaction after being taken, and the safety meets the requirement.
Test example 3
Evaluation of effect on relieving constipation-predominant irritable bowel syndrome:
the fermented soybean milks provided in application examples 1 to 3 and comparative application examples 1 to 6 were evaluated for their relieving effect on constipation-predominant irritable bowel syndrome. 100 volunteers with constipation-predominant irritable bowel syndrome (the inclusion standard is that the volunteers accord with the diagnosis standard related to irritable bowel syndrome in western medicine and Chinese medicine theories and are diagnosed, the volunteers have all typical manifestations of constipation-predominant irritable bowel syndrome, no allergic history exists in fermented soybean milk, the content of the test is known, and the test is participated in with the exclusion standard that the volunteers receive other kinds of medicine before the test begins and have cardiovascular diseases, renal diseases and other organic diseases) are randomly divided into 10 groups, 10 groups are selected, each of the 1 st to 9 th groups takes 300ml of the fermented soybean milk provided in the application examples 1 to 3 and the comparative application examples 1 to 6 every day, the 10 th group is a control group, and each of the 10 th group takes 300ml of common commercially available fermented soybean milk every day. All volunteers took a unified meal after breakfast and received a unified regular diet regimen during the trial period, including adjustment of diet structure, increase of diet salty taste and water intake, etc., and did not take other drugs for treating gastrointestinal diseases and other fermented foods. The test period was 6 months.
And (3) evaluating the curative effect: the constipation symptoms and the stool characteristics of the volunteers before and after taking the fermented soybean milk are observed, the constipation symptom score refers to a constipation symptom score table of the surgical institute of the Chinese medical society for anal and intestinal sciences group, the score table comprises 6 items of difficult defecation, stool characteristics (BSS), defecation time, feeling of incomplete tenesmus, frequency and abdominal distension, the score is set to be 0-3, and the higher the score is, the more serious the symptoms are. Symptoms of abdominal distension, abdominal pain and difficulty in defecation were integrated before and after the test in each group of volunteers
Figure BDA0003335468890000211
Are shown in Table 2. Processing data with SPSS 20.0 statistical software, testing measurement data with t test, and counting data with x2Inspection, P<0.05 indicates that there is a statistical significance for the difference. No statistical significance was observed for the differences between symptom scores in the volunteers of each group before the test (P)>0.05)
TABLE 2
Figure BDA0003335468890000212
Figure BDA0003335468890000221
Comparing the clinical effects of the groups: the curative effect is divided into cure, obvious effect, effective and ineffective. And (3) healing: the volunteers have normal defecation, normal stool texture and no abdominal distension and pain and relapse does not occur within 6 months; the effect is shown: the defecation condition and the stool quality of the volunteers are obviously improved compared with those before treatment, the defecation condition and the stool quality tend to be normal, most symptoms disappear, and relapse occurs occasionally within 6 months; the method has the following advantages: the defecation condition and the stool quality of the volunteers are improved, and the symptoms are reduced compared with those before treatment; and (4) invalidation: the defecation condition and the stool texture and the symptoms of abdominal distension and pain are not improved after the treatment of the volunteers. The results are shown in Table 3.
TABLE 3
Figure BDA0003335468890000222
Figure BDA0003335468890000231
The results show that: as can be seen from the data in tables 2 and 3, the fermented soybean milk provided in application examples 1 to 3 was significantly improved in symptoms such as abdominal distension, abdominal pain, and difficulty in defecation after being administered to the patients with constipation-predominant irritable bowel syndrome, indicating that the fermented soybean milk provided in application examples 1 to 3 has a significant effect of relieving constipation-predominant irritable bowel syndrome. In comparison with the case where none of the fermented soybean milks provided in application examples 3, 5 and 6 contains bifidobacterium lactis BLa80, the fermented soybean milks have a certain effect of improving the symptoms of abdominal distension but cannot relieve the symptoms of abdominal pain, difficulty in defecation and the like of a patient, and do not have the function of relieving constipation-predominant irritable bowel syndrome, and it is demonstrated that bifidobacterium lactis BLa80 plays a key role in regulating intestinal health and relieving constipation-predominant irritable bowel syndrome. Compared with the fermented soybean milk provided by application examples 1, 2 and 4, the fermented soybean milk has certain relieving effect on symptoms such as abdominal distension, abdominal pain, difficult defecation and the like, but the effect is obviously inferior to that of application example 1, and the synergistic effect is proved by the cooperation of three microbial agents of the streptococcus thermophilus ST81 fermented product, the lactobacillus bulgaricus LB42 fermented product and the bifidobacterium lactis BLA80 fermented product, and the effect on relieving constipation type irritable bowel syndrome is more remarkable than that of the single fermented product of the bifidobacterium lactis BLA80, the combination of the fermented product of the streptococcus thermophilus ST81 and the fermented product of the bifidobacterium lactis BLA80 and the combination of the fermented product of the lactobacillus bulgaricus LB42 and the fermented product of the bifidobacterium lactis BLA 80.
In conclusion, the invention creatively combines the fermentation product of streptococcus thermophilus ST81, the fermentation product of lactobacillus bulgaricus LB42 and the fermentation product of bifidobacterium lactis BLA80 to form the direct-administration type fermentation inoculum for relieving constipation-predominant irritable bowel syndrome. The bifidobacterium lactis BLA80 plays a key role in regulating intestinal health and relieving constipation-type irritable bowel syndrome, and is combined with streptococcus thermophilus ST81 and lactobacillus bulgaricus LB42, so that on one hand, three floras are mutually promoted, the fermentation process is facilitated, the growth of beneficial floras is promoted, on the other hand, the three floras are mutually matched, the synergistic effect is remarkable in regulating intestinal floras and relieving constipation-type irritable bowel syndrome, and the prepared fermented food can improve symptoms such as constipation and abdominal pain, so that defecation is smoother, and the constipation-type irritable bowel syndrome is effectively relieved. In addition, the invention adopts the specific combination of trehalose, malt paste, beta-cyclodextrin, sucrose, glycerol, mannitol and tween-80 as the protective agent, and has the effect of improving the stability of the prepared fermented food.
The applicant states that the present invention is described by the above examples and application examples, but the present invention is not limited to the above examples and application examples, that is, the present invention is not limited to the above examples and application examples. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the technical features described in the above embodiments can be combined in any suitable manner without contradiction, and various possible combinations of the features are not described in order to avoid unnecessary repetition.

Claims (10)

1. The direct-vat-set inoculant for relieving constipation-predominant irritable bowel syndrome comprises a streptococcus thermophilus ST81 leavening, a lactobacillus bulgaricus LB42 leavening, a bifidobacterium lactis BLA80 leavening and a protective agent.
2. The fermented product for relieving constipation-predominant irritable bowel syndrome according to claim 1, wherein the fermented product for relieving constipation-predominant irritable bowel syndrome comprises 25 to 45 parts by weight of a fermented product of streptococcus thermophilus ST81, 1.5 to 2.5 parts by weight of a fermented product of lactobacillus bulgaricus LB42, 2.5 to 65 parts by weight of a fermented product of bifidobacterium lactis BLa80, and 18 to 36 parts by weight of a protective agent;
preferably, the number of viable bacteria of bifidobacterium lactis BLA80 in the direct vat set for relieving constipation-predominant irritable bowel syndrome is 1 × 1011cfu/g or more;
preferably, the protective agent comprises any one or a combination of at least two of trehalose, maltodextrin, beta-cyclodextrin, sucrose, glycerol, mannitol or tween-80, preferably a combination of trehalose, maltodextrin, beta-cyclodextrin, sucrose, glycerol, mannitol and tween-80;
preferably, the protective agent comprises 12-18 parts of trehalose, 2.5-8.5 parts of maltodextrin, 1.0-1.5 parts of beta-cyclodextrin, 2-5 parts of sucrose, 1.2-2.4 parts of glycerol, 0.01-0.3 part of mannitol and 0.25 part of tween-800.02 in parts by weight.
3. A method for preparing the direct vat set for relieving constipation-predominant irritable bowel syndrome according to claim 1 or 2, comprising the steps of:
(1) culturing and activating streptococcus thermophilus ST81, and fermenting to obtain streptococcus thermophilus ST81 fermentation product; culturing and activating lactobacillus bulgaricus LB42, and fermenting to obtain lactobacillus bulgaricus LB42 fermentation product; culturing and activating Bifidobacterium lactis BLA80, and fermenting to obtain Bifidobacterium lactis BLA80 fermented product;
(2) and (2) mixing the streptococcus thermophilus ST81 fermentation product, the lactobacillus bulgaricus LB42 fermentation product and the bifidobacterium lactis BLA80 fermentation product obtained in the step (1) with a protective agent to obtain the direct-vat-set microbial inoculum for relieving constipation-predominant irritable bowel syndrome.
4. The method according to claim 3, wherein the temperature of the culture of Streptococcus thermophilus ST81, the temperature of the culture of Lactobacillus bulgaricus LB42 and the temperature of the culture of Bifidobacterium lactis BLA80 in step (1) are each independently 35 to 40 ℃;
preferably, the time for culturing the streptococcus thermophilus ST81, the time for culturing the lactobacillus bulgaricus LB42 and the time for culturing the bifidobacterium lactis BLA80 in the step (1) are respectively and independently 16-24 h;
preferably, the temperature of fermentation of the streptococcus thermophilus ST81, the temperature of fermentation of lactobacillus bulgaricus LB42 and the temperature of fermentation of bifidobacterium lactis BLa80 of step (1) are each independently 35-40 ℃;
preferably, the time for fermentation of the streptococcus thermophilus ST81, the time for fermentation of lactobacillus bulgaricus LB42 and the time for fermentation of bifidobacterium lactis BLa80 of step (1) are each independently 9-12 h;
preferably, the pH of the system when the streptococcus thermophilus ST81 is fermented, the pH of the system when the lactobacillus bulgaricus LB42 is fermented and the pH of the system when the bifidobacterium lactis BLA80 is fermented in the step (1) are respectively and independently 5.5-6.0;
preferably, the fermentation of the streptococcus thermophilus ST81, the fermentation of the lactobacillus bulgaricus LB42 and the fermentation of the bifidobacterium lactis BLA80 in the step (1) optionally further comprises centrifuging and collecting the fermentation products;
preferably, the rotating speed of the centrifugation is 6000-10000r/min respectively and the time of the centrifugation is 10-15min respectively and independently;
preferably, the mixing in step (2) further comprises drying;
preferably, the drying is by freeze drying.
5. The use of the fermented food product of claim 1 or 2 for relieving constipation-predominant irritable bowel syndrome in the preparation of fermented food products, wherein the amount of the fermented food product is 20 to 50 g/T.
6. The use of claim 5, wherein the fermented food product comprises any one of fermented soy milk, fermented soy flour, fermented milk, or fermented milk powder.
7. A fermented soybean milk for relieving constipation-predominant irritable bowel syndrome, which is prepared from soybean flour, a gum base, water and the direct vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome according to claim 1 or 2.
8. The fermented soybean milk for relieving constipation-predominant irritable bowel syndrome according to claim 7, wherein the raw materials for preparing the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome include, in mass percent, 0.002-0.005% of a ready-to-use fermented microbial agent for relieving constipation-predominant irritable bowel syndrome, 10-28% of soybean flour, 3-20% of a gum base, and the balance of water;
preferably, the gum base comprises any one or a combination of at least two of coconut milk, white granulated sugar, agar, pectin, citrus fiber or sea salt;
preferably, the gum base comprises coconut pulp, white granulated sugar, agar, pectin, citrus fiber, and sea salt;
preferably, the gum base material comprises, by weight, 3-10 parts of coconut milk, 1-5 parts of white granulated sugar, 0.1-1 part of agar, 0.1-0.8 part of pectin, 0.2-0.8 part of citrus fiber and 0.01-0.1 part of sea salt.
9. The preparation method of the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome according to claim 7 or 8, wherein the preparation method comprises mixing soybean flour, gum base and water, homogenizing, sterilizing, inoculating a direct-vat set microbial inoculum for relieving constipation-predominant irritable bowel syndrome, fermenting, and demulsifying to obtain the fermented soybean milk for relieving constipation-predominant irritable bowel syndrome.
10. The method of claim 9, wherein the temperature of the mixing is 60-90 ℃;
preferably, the temperature of the inoculation is 38-50 ℃;
preferably, the temperature of the fermentation is 38-50 ℃.
CN202111293299.0A 2021-11-03 2021-11-03 Direct-vat fermentation microbial inoculum for relieving constipation type irritable bowel syndrome and preparation method and application thereof Active CN114045233B (en)

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CN114146101A (en) * 2021-11-22 2022-03-08 微康益生菌(苏州)股份有限公司 Application of bifidobacterium animalis subsp lactis BLA80 in preparing medicine or food for regulating intestinal motility
CN114480208A (en) * 2022-02-18 2022-05-13 微康益生菌(苏州)股份有限公司 Direct-vat probiotic starter for relieving immune diseases and application thereof
CN115558620A (en) * 2022-09-30 2023-01-03 微康益生菌(苏州)股份有限公司 Lactic acid bacteria freeze-drying protective agent and preparation method and application thereof
CN117736943A (en) * 2024-02-20 2024-03-22 山东中科嘉亿生物工程有限公司 Composite microbial agent for improving irritable bowel syndrome and preparation method and application thereof

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CN111466440A (en) * 2020-06-03 2020-07-31 金华银河生物科技有限公司 Probiotic fermented milk capable of relieving constipation

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CN103893214A (en) * 2014-03-28 2014-07-02 北京和美科盛生物技术有限公司 Probiotics viable bacteria powder produced by whole oat solid-state mixed fermentation and preparation method of probiotics viable bacteria powder
CN111466440A (en) * 2020-06-03 2020-07-31 金华银河生物科技有限公司 Probiotic fermented milk capable of relieving constipation

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Publication number Priority date Publication date Assignee Title
CN114146101A (en) * 2021-11-22 2022-03-08 微康益生菌(苏州)股份有限公司 Application of bifidobacterium animalis subsp lactis BLA80 in preparing medicine or food for regulating intestinal motility
CN114480208A (en) * 2022-02-18 2022-05-13 微康益生菌(苏州)股份有限公司 Direct-vat probiotic starter for relieving immune diseases and application thereof
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CN115558620A (en) * 2022-09-30 2023-01-03 微康益生菌(苏州)股份有限公司 Lactic acid bacteria freeze-drying protective agent and preparation method and application thereof
CN115558620B (en) * 2022-09-30 2023-10-20 微康益生菌(苏州)股份有限公司 Lactic acid bacteria freeze-drying protective agent and preparation method and application thereof
CN117736943A (en) * 2024-02-20 2024-03-22 山东中科嘉亿生物工程有限公司 Composite microbial agent for improving irritable bowel syndrome and preparation method and application thereof
CN117736943B (en) * 2024-02-20 2024-05-24 山东中科嘉亿生物工程有限公司 Composite microbial agent for improving irritable bowel syndrome and preparation method and application thereof

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