Prebiotic composition for proliferating bifidobacterium and application thereof

Abstract

The invention discloses a prebiotic composition capable of proliferating bifidobacterium and application of the prebiotic composition. The prebiotic composition comprises polydextrose, isomaltooligosacharide and galactooligosaccharide. Wherein the mass ratio of the polydextrose to the isomaltooligosacharide to the galactooligosaccharide is (0-2): (3-5): (4-5). The prebiotic composition provided by the invention can increase the content of total bifidobacteria in intestinal tracts, especially can specifically proliferate bifidobacteria longum and bifidobacteria breve, and can accurately regulate and control intestinal flora, thereby accurately intervening in host health.

Description

Technical Field

[0001] The invention belongs to the field of food and health food, and specifically relates to a prebiotic composition for proliferating bifidobacteria and application thereof. Background Art

[0002] The International Scientific Association of Probiotics and Prebiotics (ISAPP) defines prebiotics as "a substrate selectively utilized by host microorganisms and having health benefits". Common prebiotics include inulin, oligofructose, galacto-oligosaccharide, isomaltooligosaccharide, oligoxylose, polydextrose, human milk oligosaccharide, etc. In recent years, the industry has also paid more and more attention to the research on the health benefits of prebiotics. As a functional ingredient with multiple biological activities, prebiotics provide many benefits to human health, such as regulating intestinal flora, enhancing immunity, improving nutrient absorption, etc. The physiological function of prebiotics is mainly to achieve similar functions to probiotics by promoting the growth of beneficial bacteria in the human intestine and optimizing the balance of flora. With the development of metagenomics, metabolomics, proteomics and the advancement of big data analysis methods, the concept of precise prebiotics based on individual characteristics and the clinical application of prebiotics have gradually formed, which is expected to become an important direction for the development of prebiotics in the future. Prebiotics are important functional factors in the regulation of modern intestinal function and intestinal microecology, and have become one of the means of prevention and intervention of many chronic metabolic diseases. The safety of prebiotics and their function of regulating intestinal flora have been proven by a large number of modern scientific experiments. However, due to the complexity of the entire intestinal flora, the diversity of prebiotic structure, the lack and lag of research means and methods, their development speed is relatively slow.

[0003] Among the vast intestinal bacterial community, members of the genus Bifidobacterium are among the first microorganisms to colonize the human intestine. They play an important role in the intestine from birth and are recognized as safe probiotics. Bifidobacterium is the most abundant genus in the intestine of healthy infants. In late adulthood and in a variety of diseases, the level of Bifidobacterium and its species diversity decrease. Studies have shown that obese children have lower levels of Bifidobacterium; in allergic diseases, long-term asthma patients have lower levels of Bifidobacterium; IBS patients have altered their associated microbiota compared with healthy people, and have lower levels of Bifidobacterium; the number of Bifidobacterium is reduced in elderly people with infections; and a reduction in the number of Bifidobacterium has also been observed in other diseases, such as cystic fibrosis, hepatitis B, and type I and type II diabetes. There are repeated trends between Bifidobacterium levels and different age stages and a variety of common disease states, indicating that Bifidobacterium plays an important role in human health.

[0004] The types of bifidobacteria in the intestine also change with age. Studies have found that Bifidobacterium longum, Bifidobacterium breve and Bifidobacterium bifidum are most abundant in infants; the relative abundance of Bifidobacterium breve and Bifidobacterium longum in centenarians is significantly higher than that in other populations. Bifidobacterium longum is one of the core flora members of the human intestine and one of the most abundant bifidobacteria in the human intestine. Studies have found that compared with young elderly people and young people, centenarians in Guangxi, China have significantly increased Bifidobacterium longum in the intestine. Bifidobacterium breve is a bacterium found in the human intestine. It is particularly important in the early stages of life and is one of the most abundant bacteria in the intestine of newborns. With age, Bifidobacterium breve will gradually decrease, and poor diet or lifestyle habits will lead to a large reduction in Bifidobacterium breve. Bifidobacterium breve is the dominant species in the long-lived population. Studies have found that this species has a consistent effect on potentially pathogenic intestinal bacteria. This result also supports that Bifidobacterium breve is a beneficial species that contributes to longevity. The increased abundance of Bifidobacterium longum and Bifidobacterium breve in the intestine has a certain effect on intestinal health and longevity.

[0005] At present, CN106589012A discloses a prebiotic that specifically proliferates intestinal bifidobacteria. Although it can proliferate bifidobacteria, it cannot selectively proliferate Bifidobacterium longum and Bifidobacterium breve; CN108541953A discloses a preparation method, equipment and application of a prebiotic composition that efficiently regulates intestinal flora, but it cannot selectively proliferate Bifidobacterium longum and Bifidobacterium breve; CN113862311A discloses a combination of isomerized lactose and 2'-fucosyllactose in promoting the growth of Bifidobacterium longum infantis subsp. The application in metabolic acid production can promote the growth and metabolic acid production of infant Bifidobacterium, but not Bifidobacterium longum and Bifidobacterium breve; CN101626774A discloses a composition for improving intestinal microbial flora, which can proliferate Bifidobacterium but cannot selectively proliferate Bifidobacterium longum and Bifidobacterium breve; CN114854643B discloses a culture medium for promoting the synergistic proliferation of Lactobacillus and Bifidobacterium and its application, which can proliferate Bifidobacterium but cannot selectively proliferate Bifidobacterium longum and Bifidobacterium breve. Summary of the invention

[0006] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0007] In view of the above problems and / or the problems existing in the prior art, the present invention is proposed.

[0008] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide a prebiotic composition that can proliferate total bifidobacteria in the intestine, especially can specifically proliferate Bifidobacterium longum and Bifidobacterium breve in the intestine.

[0009] In order to solve the above technical problems, the present invention provides the following technical solutions: a prebiotic composition for proliferating bifidobacteria, comprising polydextrose, isomaltooligosaccharide and galacto-oligosaccharide;

[0010] Among them, based on the mass ratio of raw materials, polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=0-2:3-5:4-5.

[0011] As a preferred embodiment of the prebiotic composition of the present invention, the ratio of the raw materials by mass is: polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=0-1:4-5:4-5.

[0012] As a preferred embodiment of the prebiotic composition of the present invention, the ratio of the raw materials by mass is: polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=1:4-5:4-5.

[0013] As a preferred embodiment of the prebiotic composition of the present invention, the prebiotic composition proliferates the total bifidobacteria in the intestinal tract of the host.

[0014] As a preferred embodiment of the probiotic composition of the present invention, the prebiotic composition has a significant effect on the specific proliferation of Bifidobacterium longum.

[0015] As a preferred embodiment of the probiotic composition of the present invention, the prebiotic composition has a significant specific proliferation effect on Bifidobacterium breve.

[0016] As a preferred embodiment of the prebiotic composition of the present invention, the host includes humans and animals.

[0017] Another object of the present invention is to overcome the deficiencies in the prior art and provide a method for preparing a prebiotic composition, comprising uniformly mixing polydextrose, isomaltooligosaccharide and galacto-oligosaccharide in a specific ratio to prepare the composition.

[0018] Another object of the present invention is to overcome the deficiencies in the prior art and provide an application of a prebiotic composition in the preparation of food and health food, wherein the food dosage forms include powders, tablets, soft candies, drinks and soft capsules.

[0019] Beneficial effects of the present invention:

[0020] (1) The prebiotic composition provided by the present invention can increase the content of total bifidobacteria in the intestine, especially can specifically increase Bifidobacterium longum and Bifidobacterium breve, accurately regulate the intestinal flora, and thus accurately intervene in the health of the host;

[0021] (2) The prebiotic combination, when polydextrose:isomaltooligosaccharide:galacto-oligosaccharide = 0-2:3-5:4-5 (more preferably, polydextrose:isomaltooligosaccharide:galacto-oligosaccharide = 0-1:4-5:4-5), has the best effect in promoting the production of short-chain fatty acids, and when utilized by intestinal flora, the gas production is small, and no obvious side effects are produced on the host intestine. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them:

[0023] Figure 1 It is a comparison chart of total bifidobacterial proliferation content in the examples of the present invention. DETAILED DESCRIPTION

[0024] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0025] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0026] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0027] The present invention is a prebiotic composition, which can be prepared into dosage forms such as powder, tablet, soft candy, beverage and the like, and can be directly orally consumed.

[0028] The composition is a composition of two or more prebiotics, and the prebiotics include polydextrose, isomaltooligosaccharide, and galacto-oligosaccharide. Among them, polydextrose: isomaltooligosaccharide: galacto-oligosaccharide = 0-2: 3-5: 4-5. Above or below this range, the proliferation of total bifidobacterium content, the specific proliferation of long bifidobacterium and breve bifidobacterium, the content of short-chain fatty acids and the gas production cannot be optimized.

[0029] The polydextrose, isomaltooligosaccharide and galacto-oligosaccharide mentioned in the embodiments of the present invention are all common commercially available products.

[0030] The Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium adolescentis, and Bifidobacterium infantis used in this embodiment are commercially available probiotic powders. The probiotic powders in this embodiment are from Jiangsu Weikang Biotechnology Co., Ltd. and Shanghai Nord Biotechnology Co., Ltd., but in principle other commercially available probiotic powders may also be used.

[0031] Example 1

[0032] Bifidobacterium animalis subsp. lactis (abbreviated as "B. lactis"), Bifidobacterium bifidum, Bifidobacterium longum subsp. longum (abbreviated as "B. longum"), Bifidobacterium breve, Bifidobacterium adolescentis, Bifidobacterium longum subsp. infantis (abbreviated as "B. infantis") are isolated, purified, identified, activated and then inoculated.

[0033] Specifically, six kinds of probiotics were inoculated into PYG medium (commercially available), cultured in an anaerobic workstation for 48-72 hours, and after activation, 200ul of bacterial solution was spread on a solidified PYG medium plate and cultured for 48-72 hours. A single colony was taken with a sterile inoculation loop and transferred to a new plate for streaking to obtain a single colony, and then a second streaking was performed for purification. Finally, a single colony was picked and transferred to PYG liquid medium for enrichment culture. After the culture was completed, the bacterial solution was collected for strain identification for subsequent experiments.

[0034] After the six species of Bifidobacterium were cultured to the logarithmic phase, 5 concentration (inoculation ratio of 1:1:1:1:1:1) was inoculated into CDM fermentation vials and placed in a constant temperature incubator at 37°C for 48 h;

[0035] Prebiotics with different proportions (see Table 2) were added as carbon sources. The concentration of prebiotics was 1%, i.e., 10 g / L. The prebiotics included galacto-oligosaccharides (GOS), isomaltooligosaccharides (IMO), and polydextrose (PDX).

[0036] Detection indicators: qPCR detection of changes in the composition of bifidobacteria before and after cultivation, short-chain fatty acids SCFAs, and total gas production.

[0037] qPCR detection of Bifidobacterium: The primer information of the strains is shown in Table 1.

[0038] Table 1

[0039]

[0040] The standard concentration plasmid 20ng / μL was diluted with dd water to 2ng / μL, 0.2ng / μL, 0.02ng / μL, 0.002ng / μL, 0.0002ng / μL to prepare the standard solution;

[0041] Take an appropriate amount of eight-tube strips and place them on an ice box, add 19 μL of the mixed solution to the bottom of the well, then add 1 μL of the mixed sample or standard solution to the well wall, cover or cover with a film, centrifuge for 30 seconds, and perform real-time fluorescence quantitative PCR on the machine for determination.

[0042] Detection of short-chain fatty acids SCFA: Use a 100μL microinjection needle to draw 80μL of acetic acid, 60μL of propionic acid, 40μL each of butyric acid, isobutyric acid, valeric acid, and isovaleric acid, and dissolve and dilute to 10mL of ultrapure water. Use ultrapure water for gradient dilution when using. Prepare standard curves for acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, and valeric acid. Take 500μL of sample extract (or fermentation broth) and place it in a centrifuge tube, add 100μL of short-chain fatty acid pretreatment solution, freeze at -30℃ for 24h, centrifuge at 10000rpm and 4℃ for 3min after thawing, take the supernatant, filter it with a filter, take 100μL of sample extract and add it to the intubation tube in the gas phase sample bottle, cover it tightly with a lid to remove bubbles, and analyze it on a gas chromatograph. Use the peak area internal standard method for quantitative determination.

[0043] Gas production measurement: Insert a barometer with a needle into the rubber stopper of the fermentation bottle. The barometer will display the pressure inside the bottle. After the value stabilizes, record the gas production.

[0044] For a comparison of total bifidobacterial proliferation content, see Figure 1 .

[0045] Table 2 Ratios of prebiotics in different groups

[0046]

[0047] Note: “ / ” indicates not measured

[0048] It can be seen that groups 4, 11 and 12 are the best; groups 7 and 9 are the second best.

[0049] Comparative Example 1

[0050] The galacto-oligosaccharide (GOS) in group 11 in Example 1 was replaced with inulin (Inulin) in equal proportion, and the effect of the prebiotic combination on the proliferation of bifidobacteria was tested by the above experimental method, and relevant indicators were detected. The results are shown in Table 3 below.

[0051] Table 3

[0052]

[0053]

[0054] The prebiotic composition of Comparative Example 1 could not proliferate total bifidobacteria well, let alone specifically proliferate Bifidobacterium breve, and its gas production was significantly higher than that of the test group in Example 1.

[0055] Prebiotics have glycosidic bonds in their molecular structure that cannot be decomposed by gastrointestinal digestive enzymes, and cannot be decomposed and utilized by human intestinal digestive enzymes, but can be used as a carbon source for probiotics, fermented and utilized by beneficial bacteria in the intestines, and promote the proliferation of probiotics; different prebiotics have different effects on the proliferation of different probiotics due to their different bond types and polymerization degrees. When the prebiotic composition of polyglucose: oligomalto-oligosaccharide: oligogalactose = 0-2: 3-5: 4-5, its structural unit composition and glycosidic bond link composition can promote bifidobacteria, especially long bifidobacterium and breve bifidobacterium, to more efficiently decompose and utilize the glycosidic bonds of the prebiotic composition, shorten their growth lag period, quickly enter a rapid growth state, increase the growth rate and bacterial concentration, produce more short-chain fatty acids, and generate less gas.

[0056] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the present invention.

Claims

1. A prebiotic composition for proliferating bifidobacteria, characterized in that: Including, polydextrose, isomaltooligosaccharide and galacto-oligosaccharide; Among them, based on the mass ratio of raw materials, polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=0-2:3-5:4-5.

2. The prebiotic composition according to claim 1, characterized in that: Calculated by the mass ratio of raw materials, polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=0-1:4-5:4-5.

3. The prebiotic composition according to claim 1 or 2, characterized in that: Calculated by the mass ratio of raw materials, polydextrose:isomaltooligosaccharide:galacto-oligosaccharide=1:4-5:4-5.

4. The prebiotic composition according to claim 3, characterized in that: The prebiotic composition proliferates total bifidobacteria in the intestinal tract of the host.

5. The prebiotic composition according to claim 4, characterized in that: The prebiotic composition specifically proliferates Bifidobacterium longum in the intestinal tract of the host.

6. The prebiotic composition according to claim 5, characterized in that: The prebiotic composition specifically proliferates Bifidobacterium breve in the intestinal tract of the host.

7. The prebiotic composition according to claim 5 or 6, characterized in that: The host includes humans and animals.

8. The method for preparing the prebiotic composition according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: uniformly mixing polydextrose, isomaltooligosaccharide and galacto-oligosaccharide in proportion to prepare the composition.

9. Use of the prebiotic composition according to any one of claims 1 to 7 in the preparation of food and health food.

10. The use according to claim 9, characterized in that: The food dosage forms include powders, tablets, soft candies, drinks and soft capsules.

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