Composition for promoting proliferation of lactic acid bacteria and application thereof

Abstract

The invention discloses a composition for promoting proliferation of lactic acid bacteria and application thereof.The composition comprises sodium hyaluronate and tremella polysaccharide, the composition has the synergistic interaction capacity on proliferation of the lactic acid bacteria by combining the sodium hyaluronate and the tremella polysaccharide, and the composition can promote proliferation of the lactic acid bacteria. On the one hand, the probiotic powder can be used as a substitute raw material of common matching prebiotics of lactic acid bacteria products, the differentiation of probiotic products in the market is improved, and the product capacity is improved; on the other hand, competitive power of terminal products is different, and reproduction and metabolism of lactic acid bacteria can be promoted; besides, the cultured lactic acid bacteria product or lactic acid bacteria, tremella polysaccharide and sodium hyaluronate can be directly applied to a subject, the intestinal flora of the subject can be balanced, and the effects of beautifying, moisturizing, protecting joints and the like can be achieved.

Description

Technical Field

[0001] The present application relates to the field of food biotechnology, and in particular to a composition for promoting the proliferation of lactic acid bacteria and its use. Background Art

[0002] The human intestine is a diverse and vibrant micro-ecosystem, home to trillions of microorganisms, including bacteria, fungi, etc., which together make up our intestinal flora. Studies have found that the number of these microorganisms far exceeds the number of our own cells. They not only play an important role in host metabolic activities and immune regulation, but are also important participants in the occurrence and defense of diseases. Many factors such as eating habits, exercise, drug (antibiotic) use, surgery, etc. can affect the intestinal flora. Therefore, it is crucial to maintain the balance of intestinal flora, and scientists are also actively looking for beneficial microorganisms to help improve human health.

[0003] Probiotics are a type of microorganisms that have beneficial effects on the human body when ingested alive. The United Nations Food and Agriculture Organization and the World Health Organization officially define probiotics as "live microorganisms that can bring health benefits to the host when ingested in sufficient quantities." At present, the most common probiotics, that is, traditional probiotic lactic acid bacteria, are mainly distributed in food, the human digestive tract, reproductive tract and our body surface. Lactic acid bacteria refer to a type of bacteria that use sugar to produce lactic acid but do not produce spores and are Gram-positive, mainly including lactobacilli and bifidobacteria.

[0004] Hyaluronic acid (HA) is a disaccharide unit structure composed of D-glucuronic acid and N-acetylglucosamine connected by β-1,3-glycosidic bonds, and then connected by β-1,4-glycosidic bonds to form a linear non-sulfated glycosaminoglycan. HA is widely distributed in the extracellular matrix and plays important physiological functions such as cell maintenance, water retention, and nutrient transport. HA can serve as a carbon source for beneficial bacteria and a substrate for peptidoglycan, the main component of the cell wall of beneficial bacteria, to maintain the balance of intestinal microecology. On January 7, 2021, the National Health Commission issued a document approving sodium hyaluronate (i.e. hyaluronic acid) as a "new food raw material" that can be used in ordinary food additives. HA is stable, odorless, soluble in water, highly safe, suitable for long-term use, and has achieved industrial production. Food-grade hyaluronic acid is a natural sodium hyaluronate with high safety and can be eaten for a long time. It can increase the water storage capacity of the skin. Oral sodium hyaluronate (food grade) is undoubtedly a better choice.

[0005] Tremella polysaccharide is an acidic heteropolysaccharide obtained from Tremella fruiting bodies. Its main chain structure is mannan connected by α-(1→3) glycosidic bonds, and its side chains are xylose, fucose, glucuronic acid, and glucose. It is a polysaccharide with a unique spatial network structure, and its molecular weight can reach more than 3 million Daltons. After Tremella polysaccharide is fermented, the glucuronic acid contained in the molecular structure is released. Glucuronic acid, as a precursor substance for the synthesis of hyaluronic acid in the body, can participate in the synthesis of HA in the body, maintain and increase the content of HA in the body, and achieve the effects of improving skin nutrition metabolism, making the skin soft, smooth, wrinkle-removing, increasing elasticity and preventing aging. Summary of the invention

[0006] Chinese patent application CN113907366 A provides an application of turmeric polysaccharide in regulating the proliferation of probiotics. By combining the methods of precooling turmeric, boiling with hot water, ethanol precipitation, and freezing and thawing, turmeric polysaccharide is extracted from turmeric tubers, which has a significant promoting effect on the proliferation of Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus delbrueckii subspecies bulgaricus, and Streptococcus thermophilus. However, this method focuses on the preparation method of turmeric polysaccharide, and does not conduct collaborative research on sodium hyaluronate and Tremella polysaccharide.

[0007] Chinese patent application CN114470005 A provides a composition of probiotics and sodium hyaluronate and its application, which is used to promote the balance of human microecology and achieve anti-inflammatory, anti-infection, laxative, anti-aging and immune-enhancing effects by combining a probiotic complex including Lactobacillus plantarum, Bifidobacterium lactis, etc. with one or more of prebiotics, double rose powder, Poria powder, vitamin C, niacin, and γ-aminobutyric acid. However, this method does not explain and study the synergistic promoting effect of sodium hyaluronate and Tremella polysaccharide with probiotics.

[0008] Chinese patent application CN112226429B discloses a method for preparing probiotic microcapsules, including: obtaining a mixed bacterial suspension; adding prebiotics, natural polymers, and sodium alginate to the mixed bacterial suspension, and forming microcapsules through emulsification and solidification steps; the prepared microcapsules have good moisturizing, acne removal, and skin microecological balance effects, and have good stability, and can be used with solvents or used as cosmetic ingredients in the cosmetics field. It is mentioned that hyaluronic acid and Tremella polysaccharide are one of the natural polymers, but the synergistic promotion effect of sodium hyaluronate and Tremella polysaccharide with probiotics has not been studied.

[0009] In order to solve the above technical problems, the present application provides a composition for promoting the proliferation of lactic acid bacteria, wherein the composition comprises sodium hyaluronate and tremella polysaccharide. Sodium hyaluronate and tremella polysaccharide act synergistically to increase the proliferation of lactic acid bacteria.

[0010] The specific technical solutions of this application are as follows:

[0011] 1. A composition for promoting the proliferation of lactic acid bacteria, comprising sodium hyaluronate and tremella polysaccharide.

[0012] 2. The composition according to item 1, wherein the mass ratio of sodium hyaluronate to Tremella polysaccharide is 1:1.25-2.5.

[0013] 3. The composition according to item 1 or 2, wherein the molecular weight of the sodium hyaluronate is 50k-400k Da, preferably 80k-110k Da.

[0014] 4. The composition according to any one of items 1 to 3, wherein the lactic acid bacteria are selected from one or more of Lactobacillus plantarum, Bifidobacterium lactis, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus casei, Lactobacillus reuteri, Lactobacillus paracasei and Bifidobacterium bifidum.

[0015] 5. Use of the composition according to any one of items 1 to 4 as a prebiotic and / or as a carbon source in promoting the proliferation of lactic acid bacteria.

[0016] 6. Use of sodium hyaluronate and Tremella polysaccharide in promoting the proliferation of lactic acid bacteria.

[0017] 7. The use according to item 6, wherein the sodium hyaluronate and Tremella polysaccharide are used as prebiotics and / or as carbon sources in promoting the proliferation of lactic acid bacteria.

[0018] 8. The use according to item 6 or 7, wherein the mass ratio of sodium hyaluronate to Tremella polysaccharide is 1:1.25-2.5.

[0019] 9. The use according to any one of items 6 to 8, wherein the molecular weight of the sodium hyaluronate is 50k-400kDa, more preferably 80k-110kDa.

[0020] 10. A culture medium product for culturing lactic acid bacteria, comprising the composition according to any one of items 1 to 6 and a culture medium.

[0021] 11. A food or health product comprising the composition according to any one of items 1 to 6 or the culture medium product according to item 10 and lactic acid bacteria.

[0022] Effects of the Invention

[0023] The composition described in the present application combines hyaluronic acid and tremella polysaccharide to have the ability to synergistically enhance the proliferation of lactic acid bacteria. On the one hand, it can be used as an alternative raw material for commonly used prebiotics in lactic acid bacteria products to enhance the differentiation of probiotic products in the market and improve product strength; on the other hand, it has a difference in the competitiveness of the terminal products, which can promote the reproduction and metabolism of lactic acid bacteria; in addition, the cultured lactic acid bacteria product or the lactic acid bacteria, tremella polysaccharide and sodium hyaluronate can be directly applied to the subject, which can balance the intestinal flora of the subject and also have the effects of beauty, moisturizing, and protecting joints.

[0024] The composition described in the present application has the advantages that the raw materials are natural, safe and reliable, safe in storage, transportation and application, stable in quality, and can be preserved for a long time. DETAILED DESCRIPTION

[0025] The embodiments described below are described in detail for the present application. Although specific embodiments of the present application are shown, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0026] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the functional differences of components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of the present application shall be determined by the attached claims.

[0027] The present application provides a composition for promoting the proliferation of lactic acid bacteria, which comprises sodium hyaluronate and tremella polysaccharide. In some embodiments, the mass ratio of sodium hyaluronate to tremella polysaccharide is 1:1.25-2.5.

[0028] For example, the mass ratio (m 透明质酸钠 :m 银耳多糖 ) can be 1:1.25, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, etc.

[0029] In some embodiments, the molecular weight of the sodium hyaluronate is 50k-400k Da. In some embodiments, the molecular weight of the sodium hyaluronate is 80k-110k Da.

[0030] For example, the molecular weight of the sodium hyaluronate can be 80k Da, 90k Da, 100k Da, 110k Da, 120k Da, 130k Da, 140k Da, 150k Da, 160k Da, 170k Da, 180k Da, 190k Da, 200k Da, 210k Da, 220k Da, 230k Da, 240k Da, 250k Da, 260k Da, 270k Da, 280k Da, 290k Da, 300k Da, 310k Da, 320k Da, 330k Da, 340k Da, 350k Da, 360k Da, 370k Da, 380k Da, 390k Da, 400k Da, etc.

[0031] In some embodiments, the lactic acid bacteria are selected from one or more of Lactobacillus plantarum, Bifidobacterium lactis, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus casei, Lactobacillus reuteri, Lactobacillus paracasei and Bifidobacterium bifidum.

[0032] In some embodiments, the composition comprises sodium hyaluronate and Tremella polysaccharide. In some embodiments, the mass ratio of the sodium hyaluronate and Tremella polysaccharide is 1: 1.25-2.5. In some embodiments, the molecular weight of the sodium hyaluronate is 50k-400k Da. In some embodiments, the molecular weight of the sodium hyaluronate is 80k-110k Da. In some embodiments, the lactic acid bacteria is selected from one or more of plant lactobacillus, bifidobacterium lactis, lactobacillus crispatus, lactobacillus acidophilus, lactobacillus rhamnosus, lactobacillus gasseri, lactobacillus casei, lactobacillus reuteri, lactobacillus paracasei and bifidobacterium bifidum.

[0033] The composition described in the present application can significantly increase the proliferation of lactic acid bacteria through the synergistic effect of sodium hyaluronate and Tremella polysaccharide.

[0034] The present application provides the use of the above-mentioned composition as a prebiotic and / or as a carbon source in promoting the proliferation of lactic acid bacteria.

[0035] The above-mentioned composition is added to the lactic acid bacteria liquid culture medium as a prebiotic and / or as a carbon source to promote the proliferation of lactic acid bacteria, which lays a foundation for the development and application of sodium hyaluronate and tremella polysaccharide in the field of lactic acid bacteria products.

[0036] The present application provides the use of sodium hyaluronate and Tremella polysaccharide as prebiotics and / or as carbon sources in promoting the proliferation of lactic acid bacteria.

[0037] The present application also provides the use of sodium hyaluronate and Tremella polysaccharide in promoting the proliferation of lactic acid bacteria. In some embodiments, the sodium hyaluronate and Tremella polysaccharide are used as prebiotics and / or as carbon sources in promoting the proliferation of lactic acid bacteria.

[0038] In some embodiments, the mass ratio of the sodium hyaluronate to the Tremella polysaccharide is 1:1.25-2.5. In some embodiments, the molecular weight of the sodium hyaluronate is 50k-400k Da, more preferably 80k-110k Da.

[0039] For example, the mass ratio (m 透明质酸钠 :m 银耳多糖 ) can be 1:1.25, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2.0, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, etc.

[0040] For example, the molecular weight of the sodium hyaluronate can be 80k Da, 90k Da, 100k Da, 110k Da, 120k Da, 130k Da, 140k Da, 150k Da, 160k Da, 170k Da, 180k Da, 190k Da, 200k Da, 210k Da, 220k Da, 230k Da, 240k Da, 250k Da, 260k Da, 270k Da, 280k Da, 290k Da, 300k Da, 310k Da, 320k Da, 330k Da, 340k Da, 350k Da, 360k Da, 370k Da, 380k Da, 390k Da, 400k Da, etc.

[0041] The present application provides a culture medium product for in vitro culture of lactic acid bacteria, which comprises the above-mentioned composition and culture medium. In the present application, the culture medium is a conventional culture medium for culturing lactic acid bacteria known in the art.

[0042] In some embodiments, the concentration of sodium hyaluronate in the culture medium product is 0.1-0.4%; and / or the concentration of Tremella polysaccharide in the culture medium product is 0.125-0.5%.

[0043] For example, the concentration of sodium hyaluronate in the culture medium product can be 0.1%, 0.2%, 0.3%, 0.4%, etc.;

[0044] The concentration of the Tremella polysaccharide in the culture medium product can be 0.125%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, etc.

[0045] The present application provides a food or health product, which comprises the above-mentioned composition or the above-mentioned culture medium product and lactic acid bacteria.

[0046] The food or health product described in the present application can be obtained by adding lactic acid bacteria, sodium hyaluronate and tremella polysaccharide to a culture medium for cultivation and then administered to a subject. The lactic acid bacteria, sodium hyaluronate and tremella polysaccharide can also be directly administered to a subject to allow the lactic acid bacteria to proliferate in the subject, thereby balancing the intestinal flora of the subject.

[0047] Example

[0048] The present application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, unless otherwise specified, % means wt%, i.e., weight percentage. The reagents or instruments used without indicating the manufacturer are all conventional reagent products that can be obtained commercially.

[0049] Example

[0050] (1) After dissolving the MRS broth medium, add sodium hyaluronate or Tremella polysaccharide or a mixture of Tremella polysaccharide and sodium hyaluronate in Table 1, and sterilize at high temperature at 121° C. for 20 min, wherein the concentrations of sodium hyaluronate and Tremella polysaccharide in Table 1 refer to the concentrations in the culture medium.

[0051] (2) According to 10 7 The lactic acid bacteria (Lactobacillus paracasei, purchased from Huaxi Biology, batch number 2022060901) were inoculated into the corresponding culture medium and cultured at 37°C for 24 h. The OD600nm value was then detected using a spectrophotometer. Three parallel experiments were performed, and the experimental data were expressed as mean ± standard deviation (mean ± SD). Each experiment was repeated at least three times independently, and GraphPad Prism and Excel were used to draw charts. The experimental data were statistically analyzed using one-way analysis of variance, and the results are shown in Table 2.

[0052] Table 1

[0053] serial number Sodium hyaluronate % Tremella polysaccharide % Proportion Control group / / / 1 0.2(10W) / / 2 0.2(40W) / / 3 0.2(100W) / / 4 0.1(10W) / / 5 0.4(10W) / / 6 / 0.125 / 7 / 0.5 / 8 0.1(10W) 0.125 1:1.25 9 0.1(10W) 0.5 1:5 10 0.2(10W) 0.125 1:0.625 11 0.2(10W) 0.5 1:2.5 12 0.4(10W) 0.125 1:0.3125 13 0.4(10W) 0.5 1:1.25

[0054] Table 2

[0055] serial number Multiplication times Relative to the control group Control group 76.93±0.15 / 1 78.86±0.73* 1.93 2 78.07±0.56* 1.14 3 / / 4 76.40±0.09 -0.53 5 77.95±0.46* 1.02 6 79.94±0.09** 3.01 7 80.45±0.09** 3.52 8 82.48±0.24** 5.55 9 76.87±0.31 -0.06 10 81.99±0.09** 5.06 11 83.44±0.09** 6.51 12 77.66±0.22* 0.73 13 82.67±0.23** 5.76

[0056] *Compared with the blank control group, P<0.05; **Compared with the blank control group, P<0.01

[0057] As can be seen from Table 2, compared with the control group, after 24 hours of culture, the proliferation multiples of lactic acid bacteria were significantly increased by adding a certain amount of sodium hyaluronate or Tremella polysaccharide (see numbers 1-2 and 4-7), indicating that these two ingredients can have a significant proliferation effect on lactic acid bacteria when added in a certain amount.

[0058] For groups numbered 1-3, experiments were conducted using sodium hyaluronate with different molecular weights. It can be seen that hyaluronic acid with different molecular weights has different effects on the proliferation of probiotics. Among them, the effect of low molecular weight 10W is better than that of medium molecular weight 40W, and the 100W molecular weight has a large viscosity, so no reference data can be obtained during the experiment. Therefore, sodium hyaluronate with a molecular weight of 10W was used for synergistic experiments in the future.

[0059] In addition, No. 8 is equivalent to the combination of No. 4 and No. 6. The increment of No. 8 relative to the control group is 5.55, which is greater than the sum of the increments of No. 4 and No. 6 relative to the control group (-0.53+3.01=2.48) (the increment of No. 8 relative to the control group is extremely significantly higher than the sum of the increments of No. 4 and No. 6 relative to the control group, p<0.01);

[0060] No. 11 is equivalent to the combination of No. 1 and No. 7. The increment of No. 11 relative to the control group is 6.51, which is greater than the sum of the increments of No. 1 and No. 7 relative to the control group (1.93+3.52=5.45) (the increment of No. 11 relative to the control group is significantly higher than the sum of the increments of No. 1 and No. 7 relative to the control group, p<0.05);

[0061] No. 13 is equivalent to the combination of No. 5 and No. 7. The increase of No. 13 relative to the control group is 5.76, which is greater than the sum of the increases of No. 5 and No. 7 relative to the control group (1.02+3.52=4.54) (the increase of No. 8 relative to the control group is significantly higher than the sum of the increases of No. 4 and No. 6 relative to the control group, p<0.05).

[0062] The ratio of sodium hyaluronate and Tremella polysaccharide used in No. 8, No. 11 and No. 13 is 1:1.25-2.5, which has a synergistic effect on the proliferation of probiotics.

[0063] In summary, sodium hyaluronate and Tremella polysaccharide have a synergistic effect at a specific ratio (1:1.25-2.5).

[0064] Therefore, sodium hyaluronate and Tremella polysaccharide can be used as a prebiotic combination for the proliferation of lactic acid bacteria and extended to products, providing ideas for solving the homogeneity of probiotic products.

[0065] The above is only the preferred embodiment of the present application, and does not limit the present application in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present application without departing from the technical solution of the present application still falls within the protection scope of the technical solution of the present application.

Claims

1. A composition for promoting the proliferation of lactic acid bacteria, comprising sodium hyaluronate and tremella polysaccharide.

2. The composition according to claim 1, wherein The mass ratio of the sodium hyaluronate to the tremella polysaccharide is 1:1.25-2.

5.

3. The composition according to claim 1 or 2, wherein The molecular weight of the sodium hyaluronate is 50k-400k Da, preferably 80k-110k Da.

4. The composition according to any one of claims 1 to 3, wherein The lactic acid bacteria are selected from one or more of Lactobacillus plantarum, Bifidobacterium lactis, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus casei, Lactobacillus reuteri, Lactobacillus paracasei and Bifidobacterium bifidum.

5. Use of the composition according to any one of claims 1 to 4 as a prebiotic and / or as a carbon source in promoting the proliferation of lactic acid bacteria.

6. Use of sodium hyaluronate and Tremella polysaccharide in promoting the proliferation of lactic acid bacteria.

7. The use according to claim 6, wherein The sodium hyaluronate and tremella polysaccharide are used as prebiotics and / or as carbon sources in promoting the proliferation of lactic acid bacteria.

8. The use according to claim 6 or 7, wherein The mass ratio of sodium hyaluronate to Tremella polysaccharide is 1:1.25-2.5; Preferably, the molecular weight of the sodium hyaluronate is 50k-400k Da, more preferably 80k-110k Da.

9. A culture medium product for culturing lactic acid bacteria, comprising the composition according to any one of claims 1 to 6 and a culture medium.

10. A food or health product comprising the composition according to any one of claims 1 to 6 or the culture medium product according to claim 9 and lactic acid bacteria.