CN112741334B - Prebiotic compositions, and uses thereof - Google Patents
Prebiotic compositions, and uses thereof Download PDFInfo
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- CN112741334B CN112741334B CN201911036025.6A CN201911036025A CN112741334B CN 112741334 B CN112741334 B CN 112741334B CN 201911036025 A CN201911036025 A CN 201911036025A CN 112741334 B CN112741334 B CN 112741334B
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- bifidobacterium animalis
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- oligosaccharide
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Abstract
The invention belongs to the technical field of food or medicine, and particularly relates to a prebiotic composition, which comprises the following components: 0.5-18 parts of oligosaccharide and 1 part of isomerized lactose; wherein the oligosaccharide is at least one selected from galacto-oligosaccharide, fructo-oligosaccharide and raffinose. The invention also relates to a (synbiotic) composition containing the prebiotic composition and bifidobacterium animalis, and applications thereof. The prebiotic composition can obviously promote the proliferation of BB12, and can be used for preventing or treating diarrhea, intestinal flora disorder caused by antibiotics, and intestinal flora disorder or respiratory infection diseases caused by iron supplement.
Description
Technical Field
The invention belongs to the technical field of food or medicine, and particularly relates to a prebiotic composition, a composition containing the prebiotic composition and animal bifidobacteria, and food or medicine application of the prebiotic composition and the composition.
Background
World Food and Agriculture Organization (FAO) and World Health Organization (WHO) on probioticsThe definition of bacteria is: live bacteria which exert an effective effect on the health of the consumer can be ingested in an appropriate amount. Among many probiotics, bifidobacteria have been attracting attention. A number of studies have shown that bifidobacteria are closely related to human life health and disease, as shown in FIG. 1 [1] : in the early life stage, the bifidobacteria in intestinal bacteria of a human body account for about 60-70 percent, and the deficiency of the bifidobacteria can cause the newborn to suffer from diseases such as necrotizing enterocolitis, allergy, IBS (IBS-induced bowel disease) and the like; in early adulthood, the bifidobacteria in intestinal bacteria of human body account for about 30-40%, and the bifidobacteria level gradually decreases along with the increase of the age in adulthood, and the lack of the bifidobacteria can cause diseases such as obesity, diabetes and the like; in the elderly, the level of bifidobacteria in the human intestinal tract is further reduced, accounting for about 0-5% of the intestinal bacteria, and the lack of bifidobacteria may lead to cancer and geriatric diseases. The number of bifidobacteria in the intestinal tract of healthy individuals and long-lived people is relatively high. It has been shown that the proportion of bifidobacteria in the faeces of formula-fed infants is significantly lower than that of breast-fed infants, which has a significant effect on the early intestinal micro-ecology of infants, and that by adding bifidobacteria probiotics to infant formula it is possible to adjust the intestinal micro-ecology of formula-fed infants closer to that of breast-fed infants, especially the proportion of bifidobacteria. Bifidobacteria are also associated with a number of potential health-promoting metabolites, including short-chain fatty acids, conjugated linoleic acid and bacteriocins.
Bifidobacterium animalis, bifidobacterium lactis BB-12, which is the most well-documented Bifidobacterium worldwide, is described in over 300 scientific publications, of which over 130 publications relate to human clinical studies, the complete genomic sequence of BB-12 has been drawn and published, numerous in vitro tests have established the strain characteristics and mechanisms of BB-12, BB-12 exhibits excellent acid bile tolerance, contains bile salt hydrolase and has strong adhesion, and these are valuable probiotic properties. The health-care effects of BB-12 were validated in many clinical studies of gastrointestinal health and immune function, which demonstrated: BB-12 survives through the gastrointestinal tract [2-4] And can maintain the health of the gastrointestinal flora, and BB-12 can maintain the balance and health of the intestinal microecologyTreating diarrhea, including diarrhea caused by antibiotic therapy [5-7] (ii) a In the aspect of immune function, clinical researches show that BB-12 can improve the resistance of an organism to common respiratory tract infection and reduce the incidence rate of acute respiratory tract infection, and the experiments prove that BB-12 inhibits pathogenic bacteria, enhances barrier function and interacts with immunity [2,8-12] . BB-12 has high stability in food, and the stability of freeze-dried powder prepared from the BB-12 is also good. For healthy people, including term newborns, BB-12 is considered safe as a dietary ingredient and dietary supplement in food products.
Prebiotics are a class of carbohydrates that are not digestible by the human body and interact with the intestinal flora in the host's gut to produce substances or effects that are beneficial to the host. The prebiotics currently under investigation include galacto-oligosaccharides, fructo-oligosaccharides, inulin, isomerized lactose, polydextrose, xylo-oligosaccharides, and the like.
The galacto-oligosaccharide (GOS) has a structure in which 1 to 7 galactosyl groups are bonded to galactosyl groups of lactose mainly through beta-1, 4-glycosidic bonds, and is obtained by transglycosidation conversion of beta-D-galactosidase with disaccharide lactose as a reaction substrate, and is a hetero-oligosaccharide formed by combining galactose and glucose. The glucosyl and galactosyl in the GOS structure are connected by beta-1, 4-glycosidic bond, and the adjacent galactosyl is connected by beta-1, 3, beta-1, 4 and beta-1, 6-glycosidic bond, wherein the beta-1, 4-glycosidic bond is taken as the main bond, and the main bonds comprise galacto-oligosaccharides, tetrasaccharides, a small amount of pentasaccharides, hexasaccharides and the like. In addition, the beta-D-galactosidase of some microorganisms can catalyze the synthesis of galactooligosaccharides structurally different from lactose, linked by beta-1, 3 and beta-1, 6-glycosidic bonds, from glucose and galactose produced by hydrolysis of the substrate lactose. Animal milk contains a small amount of GOS, and human breast milk contains a little more. The structure of galacto-oligosaccharides (GOS) is shown by the following formula:
molecular structure of Galactooligosaccharides (GOS).
The fructo-oligosaccharide is functional oligosaccharide with beta (2-1) glycosidic bond connection and polymerization degree of 2-9, belongs to food ingredients, and is widely present in fruits, vegetables and other substances. According to different production raw materials and processes, the fructo-oligosaccharide can be divided into two types on the whole, one type is inulin fructo-oligosaccharide, the production raw material is inulin extracted from chicory or jerusalem artichoke, the inulin is generated by enzymolysis or acidolysis, the main structural form of the fructo-oligosaccharide is Fn type (F is fructose, and n is the number of the fructose), and the fructo-oligosaccharide also contains a certain amount of GFn type; the other is sucrose fructo-oligosaccharide, the production raw material is sucrose, and the sucrose fructo-oligosaccharide, tetrasaccharide, pentasaccharide, hexasaccharide and the mixture thereof are generated by the action of enzyme, and the structural form of the sucrose fructo-oligosaccharide is GFn type. The two fructooligosaccharides are slightly different in structure, but have substantially the same physiological function. The structures of GFn type and Fn type are shown as follows:
chemical structures of GFn-type (left) and Fn-type (right).
Isomerized lactose is the earliest prebiotic found, then called the "bifidus factor", and has a good promoting effect on the growth of bifidobacteria. The isomerized lactose is a disaccharide, and has various physicochemical advantages of low sweetness, good acid resistance and good heat stability.
Synbiotics generally refers to a combination of probiotics and prebiotics, wherein the prebiotics enhance the growth and reproduction ability of the probiotics, the survival ability in the intestinal system, and thus provide health benefits to the host. The top experts of the organization of ISAPP (International scientific Association for Probiotics and Prebiotics) in 2019 at 5 months are trying to demonstrate a definition of synbiotics together, but the demonstration result has not been published yet.
There is a need for a new prebiotic composition to enhance the proliferative effect of BB12.
Disclosure of Invention
The present invention provides a prebiotic composition that significantly promotes the proliferation of BB12. On the basis, the invention also provides a composition (synbiotic composition) containing the prebiotic composition and BB12, and food application and medical application of the prebiotic composition and the composition.
The invention relates to a prebiotic composition, which comprises the following components:
0.5 to 18 parts by weight of oligosaccharide (e.g., 0.6 part by weight, 0.7 part by weight, 0.8 part by weight, 0.9 part by weight, 1 part by weight, 1.5 part by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, 5.5 parts by weight, 6 parts by weight, 6.5 parts by weight, 7 parts by weight, 7.5 parts by weight, 8 parts by weight, 8.5 parts by weight, 9 parts by weight, 9.5 parts by weight, 10 parts by weight, 10.5 parts by weight, 11 parts by weight, 11.5 parts by weight, 12 parts by weight, 12.5 parts by weight, 13 parts by weight, 13.5 parts by weight, 14 parts by weight, 14.5 parts by weight, 15 parts by weight, 15.5 parts by weight, 17 parts by weight)
1 part by weight of isomerized lactose;
wherein the oligosaccharide is at least one selected from galacto-oligosaccharide, fructo-oligosaccharide and raffinose.
In some embodiments of the first aspect of the present invention, the oligosaccharide is a galactooligosaccharide and/or a fructooligosaccharide.
In certain embodiments of the first aspect of the present invention, the oligosaccharide is 1 to 4 parts by weight.
In some embodiments of the first aspect of the present invention, the prebiotic composition further comprises an additional prebiotic ingredient.
In some embodiments of the invention, the prebiotic composition further comprises at least one prebiotic component selected from the group consisting of human milk oligosaccharides, fructooligosaccharides, transgalactosyl oligosaccharides, soy oligosaccharides, isomalto-oligosaccharides, neosugars, lactitol oligosaccharides, mannooligosaccharides, gentiooligosaccharides, xylooligosaccharides, polydextrose, dextran, polyfructose, chicory, resistant dextrins, resistant starch, gum arabic, locust bean gum, and guar gum.
In some embodiments of the first aspect of the present invention, the oligosaccharide is a galactooligosaccharide; preferably, the galactooligosaccharide is 2 to 16 parts by weight, such as 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, 5.5 parts by weight, 6 parts by weight, 6.5 parts by weight, 7 parts by weight, 7.5 parts by weight, 8 parts by weight, 8.5 parts by weight, 9 parts by weight, 9.5 parts by weight, 10 parts by weight, 10.5 parts by weight, 11 parts by weight, 12 parts by weight, 13 parts by weight, 14 parts by weight, 15 parts by weight.
In some embodiments of the first aspect of the present invention, the oligosaccharide is a fructooligosaccharide; preferably, the fructooligosaccharide is 1 to 3.7 parts by weight, such as 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight.
A second aspect of the invention relates to a composition comprising a prebiotic composition according to the first aspect of the invention and a probiotic, which is bifidobacterium animalis (bifidobacterium animalis).
In some embodiments of the second aspect of the invention, the probiotic is bifidobacterium animalis BB12.
In some embodiments of the second aspect of the present invention, the composition is a synbiotic composition.
In some embodiments of the second aspect of the present invention, the composition is a synbiotic formulation.
In some embodiments of the invention, the types of synbiotic formulations include, but are not limited to, solutions, powders, tablets, granules, pills, emulsions, suspensions, drops, sprays, ointments, gels, pastes, and the like.
In some embodiments of the second aspect of the invention, the probiotic is not less than 10 per 1-30 g of prebiotic composition 8 CFU。
In some embodiments of the second aspect of the invention, the ratio of prebiotic composition to probiotic is 1 (10) 8 ～10 13 ) g/CFU, e.g. 1 9 g/CFU、1:10 10 g/CFU、1:10 11 g/CFU、1:10 12 g/CFU。
In a third aspect, the present invention relates to a method for culturing or promoting the proliferation of bifidobacterium animalis, comprising the steps of:
the composition according to the second aspect of the present invention is added to a culture medium and cultured for 6 to 30 hours (e.g., 7 hours, 8 hours, 9 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 21 hours, 23 hours, 25 hours, 27 hours, 29 hours) at 30 to 40 ℃ (e.g., 31 ℃, 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, 37 ℃, 38 ℃, 39 ℃) to obtain a culture.
In some embodiments of the third aspect of the invention, the medium is MRS +0.05% cysteine medium.
In some embodiments of the invention, MRS +0.05% cysteine medium refers to de Man Rogosa Sharpe (MRS) broth (Difco, detroit, md.) medium containing 0.05% (W/W) cysteine.
In some embodiments of the third aspect of the invention, the weight of the prebiotic composition in the composition is between 0.4% and 4% of the weight of the medium, e.g. 0.8%, 1%, 2%, 3%.
In some embodiments of the third aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In a fourth aspect, the invention relates to a culture comprising bifidobacterium animalis produced by the method of the third aspect of the invention.
In some embodiments of the fourth aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In some embodiments of the fourth aspect of the invention, the Bifidobacterium animalis in the culture has an OD600 reading of ≥ 0.38, preferably 0.4-0.5, e.g. 0.39, 0.42, 0.44, 0.45, 0.46, 0.48, 0.50, 0.52, 0.55, 0.57, 0.60, 0.70, 0.80, 0.90.
A fifth aspect of the invention relates to a food or pharmaceutical product comprising a prebiotic composition according to the first aspect of the invention and bifidobacterium animalis, or a composition according to the second aspect of the invention, or a culture comprising bifidobacterium animalis according to the fourth aspect of the invention.
In some embodiments of the fifth aspect of the invention, the food or pharmaceutical product has a bifidobacterium animalis content of not less than 10 per 1-30 g prebiotic composition 8 CFU。
In some embodiments of the fifth aspect of the invention, the ratio of prebiotic composition to Bifidobacterium animalis in the food or pharmaceutical product is 1 (10) 8 ～10 13 ) g/CFU, e.g. 1 9 g/CFU、1:10 10 g/CFU、1:10 11 g/CFU、1:10 12 g/CFU。
In some embodiments of the fifth aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In some embodiments of the fifth aspect of the present invention, the food product is selected from at least one of dairy products, cereal products, soy products, condiments, ice products and beverages, such as dairy products.
In some embodiments of the fifth aspect of the present invention, the dairy product is selected from at least one of a group consisting of a sterilized milk, a reconstituted milk, a yogurt milk, a milk powder, a milk formula, a condensed milk, a cheese, a casein, a whey powder and a milk fat, preferably a milk formula, more preferably a milk formula for infants.
A sixth aspect of the invention relates to the use of a prebiotic composition and bifidobacterium animalis in the preparation of a synbiotic formulation according to the first aspect of the invention.
In some embodiments of the sixth aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In some embodiments of the sixth aspect of the invention, the bifidobacterium animalis is not less than 10 per 1-30 g of prebiotic composition 8 CFU。
In some embodiments of the sixth aspect of the invention, the ratio of prebiotic composition to Bifidobacterium animalis is 1 (10) 8 ～10 13 ) g/CFU, e.g. 1 9 g/CFU、1:10 10 g/CFU、1:10 11 g/CFU、1:10 12 g/CFU。
The seventh aspect of the invention relates to the prebiotic composition and bifidobacterium animalis of the first aspect of the invention, or the use of the composition of the second aspect of the invention in culturing or promoting the proliferation of bifidobacterium animalis.
In some embodiments of the seventh aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In some embodiments of the seventh aspect of the invention, the bifidobacterium animalis is not less than 10 per 1-30 g of prebiotic composition 8 CFU。
In some embodiments of the seventh aspect of the invention, the ratio of prebiotic composition to Bifidobacterium animalis is 1 (10) 8 ～10 13 ) g/CFU, e.g. 1 9 g/CFU、1:10 10 g/CFU、1:10 11 g/CFU、1:10 12 g/CFU。
An eighth aspect of the present invention relates to the use of the prebiotic composition according to the first aspect of the present invention and bifidobacterium animalis, or the composition according to the second aspect of the present invention, or the culture containing bifidobacterium animalis according to the fourth aspect of the present invention in food, or in the manufacture of a medicament for the prevention or treatment of diarrhea, intestinal flora disturbance caused by antibiotics, intestinal flora disturbance caused by iron supplementation, or respiratory infection.
In some embodiments of the eighth aspect of the invention, the bifidobacterium animalis is bifidobacterium animalis BB12.
In some embodiments of the eighth aspect of the invention, the bifidobacterium animalis is not less than 10 per 1-30 g of prebiotic composition 8 CFU。
In some embodiments of the eighth aspect of the invention, the ratio of prebiotic composition to Bifidobacterium animalis is 1 (10) 8 ～10 13 ) g/CFU, e.g. 1 9 g/CFU、1:10 10 g/CFU、1:10 11 g/CFU、1:10 12 g/CFU。
In some embodiments of the invention, the galactooligosaccharides have a relative molecular mass of 300 to 2000.
In some embodiments of the present invention, the galactooligosaccharide may be a single structure galactooligosaccharide or a mixture of galactooligosaccharides with different structures.
In some embodiments of the invention, the galactooligosaccharide is selected from at least one of galactooligosaccharide, galactooligosaccharide pentasaccharide and galactooligosaccharide hexasaccharide.
In some embodiments of the invention, the galactooligosaccharide may be provided in the form of a galactooligosaccharide powder or a galactooligosaccharide solution.
In some embodiments of the invention, the galactooligosaccharide content of the dry weight of the lacto-oligosaccharide powder or galactooligosaccharide solution is greater than or equal to 57%.
In some embodiments of the invention, the galactooligosaccharide complies with the regulation of ministry of health of the people's republic of china No. 20, 2008.
In some embodiments of the present invention, the fructooligosaccharide may be inulin fructooligosaccharide or sucrose fructooligosaccharide, or a mixture thereof.
In some embodiments of the invention, the fructooligosaccharide may be of the GFn type or the Fn type alone or in combination. Wherein, the GFn type can be a single structure with n taking a fixed value, and can also be a mixture of different GFn types; the Fn type may be a single structure or a mixture of different Fn types.
In some embodiments of the invention, the fructooligosaccharides comply with the regulations of GB/T23528-2009 fructooligosaccharides.
In some embodiments of the invention, the isomerized lactose complies with the specifications in GB 1886.176-2016 for isomerized lactose.
In some embodiments of the present invention, the raffinose meets the specifications of "national food safety standard food nutrient supplement raffinose" of GB 31618-2014.
The invention has the following beneficial effects:
1. the prebiotic composition can obviously promote the proliferation of BB12.
2. The composition (synbiotic composition) of the present invention is useful for preventing or treating diarrhea, intestinal flora disturbance caused by antibiotics, intestinal flora disturbance caused by iron supplementation, or respiratory infectious diseases.
Drawings
In order that the present disclosure may be more readily and clearly understood, reference is now made to the following detailed description of the present disclosure taken in conjunction with the accompanying drawings, in which:
FIG. 1 is a graphical representation of the results of a prior study of the health and disease relationship of bifidobacteria to humans;
FIG. 2 is the results of OD600 readings for various combinations in example 1 of the present invention;
FIG. 3 shows the results of various combinations of OD600 readings in example 2 of the present invention.
Detailed Description
The embodiments of the present invention will now be described more fully hereinafter with reference to the accompanying examples, in which some, but not all embodiments of the invention are shown. The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Bifidobacterium animalis BB12 strain was supplied by Kehansen.
(1) BB12 strain was placed in de Man Rogosa Sharpe (MRS) broth (Difco, detroit, md.) supplemented with 0.5g/L of L-cysteine and incubated at 37 ℃ for 24h in an anaerobic environment to obtain a culture. The cultures were then passaged twice on semisynthetic MRS medium (sMRS) +0.5 g/L-cysteine supplemented with 1% (w/v) sterile-filtered glucose as the sole carbohydrate source; after the second passage, the resulting culture was used as an inoculum for future use. Wherein, the mother liquor of each carbon source is filtered and sterilized by a filter with the pore diameter of 0.22 micron and is used at the final concentration of 1 percent.
(2) Let Galactooligosaccharide (GOS) group, isomerized Lactose (LOS) group, GOS/LOS (weight ratio 1). Each group was inoculated with BB12 in MRS +0.05% cysteine medium at an OD600 reading of 0.1 and added with 1% Galactooligosaccharide (GOS), 1% isomerized Lactose (LOS), 1% GOS/LOS (1 by weight) 10 1CFU/g, the temperature of the culture is 35-38 ℃, OD600 in each group of culture medium is measured by High Performance Liquid Chromatography (HPLC) after 15 hours of culture,the results are shown in FIG. 2.
As can be seen from fig. 2, the proliferation amount of BB12 in the GOS/LOS (weight ratio 4. But the pH of each broth was not significantly different. This suggests that a certain proportion of GOS and LOS synergistically promote the proliferation of BB12.
Example 2
(1) Same as in step (1) of example 1.
(2) The Fructooligosaccharide (FOS) group, the isomerized Lactose (LOS) group, the FOS/LOS (1 by weight), the FOS/LOS (4 by weight. Each group was inoculated with BB12 in MRS +0.05% cysteine medium according to OD600 reading 0.1 and added with 1% Fructooligosaccharide (FOS), 1% isomerized Lactose (LOS), 1% FOS/LOS (1 in weight 10 1CFU/g, at a temperature of 35 to 38 ℃ for 15 hours, and then measured for OD600 by High Performance Liquid Chromatography (HPLC), the results are shown in FIG. 3.
As can be seen from fig. 3, the proliferation amount of BB12 in FOS/LOS (weight ratio 1. But the pH of each broth was not significantly different. This indicates that a certain proportion of FOS and LOS synergistically promote the proliferation of BB12.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications derived therefrom are intended to be within the scope of the invention.
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Claims (9)
1. A composition comprises prebiotic composition and Bifidobacterium animalis BB12, wherein the ratio of prebiotic composition to Bifidobacterium animalis BB12 is 1 (10) 8 ～10 11 ) g/CFU; wherein,
the prebiotic composition consists of the following components:
galacto-oligosaccharide 4 parts by weight
1 part by weight of isomerized lactose;
alternatively, the prebiotic composition is comprised of:
1 part by weight of fructo-oligosaccharide
Isomerized lactose 1 part by weight.
2. A method for culturing Bifidobacterium animalis BB12 or promoting the proliferation of Bifidobacterium animalis BB12, comprising the steps of:
the composition of claim 1 is added to MRS +0.05% cysteine medium, the weight of the prebiotic composition is 0.4-2% of the weight of the medium, and the culture is obtained by culturing at 30-40 ℃ for 14-16 hours.
3. A culture comprising bifidobacterium animalis BB12 produced by the method of claim 2.
4. A food or pharmaceutical product comprising the composition of claim 1 or comprising the culture of bifidobacterium animalis BB12 of claim 3.
5. The food product of claim 4, wherein the food product is selected from at least one of dairy products, cereal products, soy products, condiments, ice products, and beverages.
6. A food product according to claim 4 wherein the food product is a dairy product.
7. The food product of claim 5, wherein the dairy product is selected from at least one of the group consisting of sterilized milk, reconstituted milk, yogurt milk, powdered milk formula, condensed milk, cheese, casein, whey powder, and milk fat.
8. Use of the composition of claim 1 for culturing bifidobacterium animalis BB12 or promoting the proliferation of bifidobacterium animalis BB12.
9. Use of the composition of claim 1 or the culture containing bifidobacterium animalis BB12 of claim 3 in the preparation of a food or a medicament for the prevention or treatment of diarrhea, intestinal flora disturbance caused by antibiotics, intestinal flora disturbance caused by iron supplementation or respiratory infectious diseases.
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