A probiotic composition and its preparation method

Abstract

This application relates to the field of biological fermentation technology, and specifically discloses a probiotic composition and a preparation method thereof. The probiotic composition provided by this application comprises the following components in parts by weight: Bifidobacterium longum 5 - 15 parts, Saccharopolyspora sp. 3 - 10 parts, longan seed powder 5 - 15 parts, and small molecule peptides 2 - 6 parts; further, the probiotic composition comprises the following components in parts by weight: Bifidobacterium longum 5 - 15 parts, Saccharopolyspora sp. 2 - 4 parts, longan seed powder 8 - 13 parts, and small molecule peptides 2 - 6 parts. The probiotic composition provided by this application can effectively inhibit the growth of Helicobacter pylori and improve gastrointestinal discomfort.

Description

Technical Field

[0001] This application relates to the field of biological fermentation technology, and specifically relates to a probiotic composition and a preparation method thereof. Background Art

[0002] Helicobacter pylori is a Gram-negative, microaerophilic bacterium that mainly lives in various regions of the stomach and duodenum. When Helicobacter pylori multiplies in large numbers in the stomach, it can cause mild chronic gastritis, and in severe cases, it may lead to peptic ulcer and gastric cancer. Therefore, the current main treatment method for Helicobacter pylori is the use of antibiotics. However, long-term use of antibiotics not only causes gastrointestinal dysfunction and affects the health of the body, but also increases the drug resistance of Helicobacter pylori, making it difficult for a small amount of antibiotics to play a therapeutic role, causing serious physical and economic burdens on patients. Summary of the Invention

[0003] In order to effectively inhibit Helicobacter pylori and improve gastrointestinal discomfort, this application provides a probiotic composition and a preparation method thereof.

[0004] In a first aspect, this application provides a probiotic composition, adopting the following technical solution:

[0005] A probiotic composition includes the following components in parts by weight: 5 - 15 parts of Bifidobacterium longum, 1 - 5 parts of Saccharopolyspora, 5 - 15 parts of longan seed powder, and 2 - 6 parts of small molecule peptides.

[0006] This application uses Bifidobacterium longum, Saccharopolyspora longan and small molecule peptide powder to prepare a probiotic composition, which can effectively inhibit the growth of Helicobacter pylori and improve intestinal discomfort caused by intestinal flora imbalance. The inventors of this application found that longan seed powder has a good inhibitory effect on the vacuolating cytotoxin A protein in type I strains of Helicobacter pylori, can inhibit the growth of Helicobacter pylori, and improve gastrointestinal discomfort caused by Helicobacter pylori; Bifidobacterium longum can inhibit the growth of Helicobacter pylori, prevent the adhesion of Helicobacter pylori in the intestine, reduce the pH value of the intestine, and inhibit the growth of harmful bacteria, maintaining the balance of the intestinal flora. Related technologies indicate that Saccharopolyspora has a degradation effect on tyramine, histamine, putrescine, and cadaverine, and can be used for the fermentation of stinky mandarin fish, cooking wine, and vinegar, etc.; however, the inventors of this application found that when Bifidobacterium longum is combined with a small amount of Saccharopolyspora, the inhibitory effect of the obtained probiotic composition on Helicobacter pylori will be significantly enhanced. Therefore, through a large number of experimental explorations, this application has prepared a probiotic composition with excellent inhibitory effect on Helicobacter pylori and good gastrointestinal improvement effect.

[0007] Preferably, the probiotic composition comprises the following components in parts by weight: 5-15 parts of Bifidobacterium longum, 2-4 parts of Saccharopolyspora, 5-15 parts of longan seed powder, and 2-6 parts of small molecule peptides.

[0008] In the present application, the weight ratio of Bifidobacterium longum to Saccharopolyspora is further adjusted to be between 1:(0.2-0.4), and the obtained probiotic composition has a better inhibitory effect on Helicobacter pylori and a better improvement effect on the gastrointestinal tract.

[0009] More preferably, the probiotic composition comprises the following components in parts by weight: 5-15 parts of Bifidobacterium longum, 2-4 parts of Saccharopolyspora, 8-13 parts of longan seed powder, and 2-6 parts of small molecule peptides.

[0010] In the present application, by further adjusting the addition amount of longan seed powder, a probiotic composition with a better inhibitory effect on Helicobacter pylori and a better improvement effect on intestinal discomfort is obtained.

[0011] In some embodiments, the parts by weight of the Saccharopolyspora can be 1-2 parts, 1-4 parts, 2-4 parts, 2-5 parts, or 4-5 parts.

[0012] In a specific embodiment, the parts by weight of the Saccharopolyspora can also be 1 part, 2 parts, 4 parts, or 5 parts.

[0013] In some embodiments, the parts by weight of the longan seed powder can be 0.5-0.8 parts, 0.5-1.3 parts, 0.8-1.3 parts, 0.8-1.5 parts, or 1.3-1.5 parts.

[0014] In a specific embodiment, the parts by weight of the longan seed powder can also be 0.5 part, 0.8 part, 1.3 parts, or 1.5 parts.

[0015] In the present application, the longan seed powder can be commercially available or can be self-made by the following method.

[0016] The preparation method of the longan seed powder is as follows: Wash and dry the longan seeds, scrape off the black skin on the surface, and grind them into powder; then place the powder in a microwave device and irradiate it at 600-800W for 15-25 minutes to obtain the longan seed powder.

[0017] The inventors of the present application have found through research that after microwave irradiation of the longan seed powder, the obtained longan seed powder has a stronger inhibitory effect on Helicobacter pylori; further controlling the irradiation power and irradiation time within the above range, the obtained longan seed powder has a more obvious inhibitory effect on Helicobacter pylori and a more significant improvement effect on gastrointestinal discomfort.

[0018] In some embodiments, the irradiation power may be 500 - 600W, 500 - 700W, 500 - 800W, 600 - 700W or 700 - 800W.

[0019] In a specific embodiment, the irradiation power may also be 500W, 600W, 700W or 800W.

[0020] In some embodiments, the irradiation time may be 15 - 20 min or 20 - 25 min.

[0021] In a specific embodiment, the irradiation time may also be 15 min, 20 min or 25 min.

[0022] In a second aspect, the present application provides the use of the probiotic composition as a pharmaceutical adjuvant.

[0023] The use of the probiotic composition provided by the present application is not limited to the above pharmaceutical adjuvants; all probiotic foods for improving the intestine prepared using the probiotic composition of the present application fall within the protection scope of the present application.

[0024] In summary, the present application has the following beneficial effects:

[0025] 1. The probiotic composition of the present application is prepared from Bifidobacterium longum, Saccharopolyspora, longan seed powder and small molecule peptides. This probiotic composition can reduce the levels of inflammatory growth factors IL-6 and IL-8 and inhibit the inflammation caused by Helicobacter pylori.

[0026] 2. By subjecting the longan seed powder to microwave treatment and controlling the irradiation power at 600 - 800W and the irradiation time at 15 - 25 min, the present application can further enhance the inhibitory effect of the longan seed powder on Helicobacter pylori. Specific Embodiments

[0027] The present application provides a probiotic composition, which comprises the following components in parts by weight: 5 - 15 parts of Bifidobacterium longum, 1 - 5 parts of Saccharopolyspora, 5 - 15 parts of longan seed powder and 2 - 6 parts of small molecule peptides. Further, the probiotic composition comprises the following components in parts by weight: 5 - 15 parts of Bifidobacterium longum, 2 - 4 parts of Saccharopolyspora, 8 - 13 parts of longan seed powder and 2 - 6 parts of small molecule peptides.

[0028] Among them, the preparation method of the longan seed powder is: washing and drying longan seeds, scraping off the surface black skin, and grinding them into powder; then placing the powder in a microwave device and irradiating it at 600 - 800W for 15 - 25 min to obtain the longan seed powder.

[0029] In this application, the preparation method of the probiotic composition is as follows: Bifidobacterium longum, Saccharopolyspora sp., longan seed powder and small molecule peptides are mixed evenly according to the weight ratio to obtain the probiotic composition.

[0030] In this application, the Bifidobacterium longum is strain LTBL16 with the deposit number of CCTCC M 2019028; the Saccharopolyspora sp. is Saccharopolyspora jiangxiensis J3 with the deposit number of CCTCC NO: M 2020104; the commercially available longan seed powder is purchased from Ningxia Vanilla Biotechnology Co., Ltd.; the small molecule peptides are bovine bone marrow small molecule peptides or soybean small molecule peptides, and the bovine bone marrow small molecule peptides are purchased from Shaanxi Xiazhou Biotechnology Co., Ltd.; the other raw materials, reagents, solvents, etc. used in this application can all be obtained through commercial purchase.

[0031] The following further elaborates on this application in combination with preparation examples, examples and performance detection tests.

[0032] Preparation Examples 1-7

[0033] Preparation Examples 1-7 respectively provide a probiotic composition.

[0034] The difference in the above preparation examples lies in: the weight ratio of each component in the probiotic composition, as specifically shown in Table 1.

[0035] The preparation method of the above probiotic composition is as follows: Bifidobacterium longum, Saccharopolyspora sp., small molecule peptides and longan seed powder are mixed evenly according to the weight ratio to obtain the probiotic composition. The longan seed powder used is commercially available.

[0036] Table 1 The addition amounts of each component in the probiotic composition provided by Preparation Examples 1-7

[0037]

[0038]

[0039] Preparation Example 8

[0040] Preparation Example 8 is prepared according to the method of Preparation Example 3, with the difference that: the longan seed powder used in Preparation Example 8 is treated by microwave.

[0041] In Preparation Example 8, the preparation method of the longan seed powder is as follows: the longan seeds are washed, dried in the sun, the surface black skin is scraped off, and then ground into powder; then the powder is placed in a microwave device and irradiated at 600W for 20 min to obtain the longan seed powder.

[0042] Preparation Examples 9-13

[0043] Preparation Examples 9-13 are prepared according to the method of Preparation Example 8, with the difference that: the conditions of microwave treatment; specifically as shown in Table 2 below.

[0044] Table 2 Microwave treatment conditions of longan seed powder in Preparation Examples 9 - 13

[0045]

[0046] Examples 1 - 13

[0047] Examples 1 - 13 respectively provide a fermentation composition.

[0048] The differences among the above - mentioned examples are as follows: The probiotic compositions used in Examples 1 - 13 are respectively derived from Preparation Examples 1 - 13.

[0049] The preparation method of the above - mentioned fermentation composition includes the following steps:

[0050] (1) Ingredient preparation: Dissolve 100 g of skim milk powder in 150 mL of water at 65 °C, stir evenly until completely dissolved; then sterilize it under steam at a pressure ≥ 0.8 MPa and a temperature of 100 - 120 °C for 3 - 5 s to obtain a pre - sterilized mixture.

[0051] (2) Fermentation: Mix the pre - sterilized mixture with 1.5 g of the probiotic composition, and then ferment at 40 °C for 12 h to obtain fermented milk.

[0052] (3) Ingredient preparation: Mix 3 g of propylene glycol alginate, 0.05 g of acesulfame potassium, 0.55 g of edible essence, 1.2 g of sucralose, and 5.5 g of polydextrose evenly and dissolve them in water at 70 °C. After dissolution, add them to the fermented milk obtained in step (2), and stir evenly to obtain a secondary ingredient mixture.

[0053] (4) Acid adjustment: Add sodium citrate to the secondary ingredient mixture to adjust the acidity to 45 - 50 °T to obtain an acid - adjusted mixture.

[0054] (5) Homogenization: Preheat the secondary ingredient mixture to 60 °C, and then perform two - stage step - type homogenization; in the two - stage step - type homogenization, the pressure in the first stage is 20 MPa, and the pressure in the second stage is 5 MPa.

[0055] (6) Secondary sterilization: Subject the homogenized material to UHT ultra - high - pressure instantaneous sterilization at 120 °C for 10 - 30 s, and cool down after sterilization to obtain the fermentation composition.

[0056] Examples 14 - 17

[0057] Examples 14 - 17 are carried out according to the method of Example 8, with the difference being that the weight ratio of the probiotic composition to the skim milk powder is as shown in Table 3 specifically.

[0058] Table 3 Weight ratio of the probiotic composition to the skim milk powder in the fermentation compositions provided by Examples 14 - 17

[0059]

[0060] Example 18

[0061] Example 18 was carried out according to the method of Example 15, except that: the homogenization step in the preparation method of the fermentation composition is as follows:

[0062] (5) Homogenization: Preheat the secondary batching mixture to 60 °C, and then carry out two - stage stepped homogenization; in the two - stage stepped homogenization, the pressure of the first - stage homogenization is 25 MPa, and the pressure of the second - stage homogenization is 10 MPa.

[0063] Example 19

[0064] Example 19 was carried out according to the method of Example 15, except that: the homogenization step in the preparation method of the fermentation composition is as follows:

[0065] (5) Homogenization: Preheat the secondary batching mixture to 60 °C, and then carry out two - stage stepped homogenization; in the two - stage stepped homogenization, the pressure of the first - stage homogenization is 20 MPa, and the pressure of the second - stage homogenization is 10 MPa.

[0066] Example 20

[0067] Example 20 was carried out according to the method of Example 15, except that: the homogenization step in the preparation method of the fermentation composition is as follows:

[0068] (5) Homogenization: Preheat the secondary batching mixture to 60 °C, and then carry out two - stage stepped homogenization; in the two - stage stepped homogenization, the pressure of the first - stage homogenization is 25 MPa, and the pressure of the second - stage homogenization is 5 MPa.

[0069] Comparative Example 1

[0070] Comparative Example 1 was carried out according to the method of Example 3, except that: the probiotic composition used in the fermentation composition.

[0071] The preparation method of the probiotic composition in Comparative Example 1 is as follows: Mix 1 g of Bifidobacterium longum, 0.4 g of small - molecule peptide and 1.3 g of longan seed powder evenly by weight to obtain the probiotic composition.

[0072] Comparative Example 2

[0073] Comparative Example 2 was carried out according to the method of Example 3, except that: the probiotic composition used in the fermentation composition.

[0074] The preparation method of the probiotic composition in Comparative Example 2 is as follows: Mix 0.4 g of Saccharopolyspora, 0.4 g of small molecule peptides, and 1.3 g of longan seed powder evenly according to the weight ratio to obtain the probiotic composition.

[0075] Comparative Example 3

[0076] Perform Comparative Example 3 according to the method of Example 3, with the difference that: the probiotic composition used in the fermentation composition.

[0077] The preparation method of the probiotic composition in Comparative Example 3 is as follows: Mix 1 g of Bifidobacterium longum, 0.4 g of Saccharopolyspora, and 0.4 g of small molecule peptides evenly according to the weight ratio to obtain the probiotic composition.

[0078] Comparative Example 4

[0079] Perform Comparative Example 4 according to the method of Example 15, with the difference that: the homogenization step in the preparation method of the fermentation composition is as follows:

[0080] (5) Homogenization: Preheat the secondary batching mixture to 60 °C, and then perform two-stage stepped homogenization; in the two-stage stepped homogenization, the pressure of the first stage homogenization is 15 MPa, and the pressure of the second stage homogenization is 30 MPa.

[0081] Comparative Example 5

[0082] Perform Comparative Example 5 according to the method of Example 15, with the difference that: the homogenization step in the preparation method of the fermentation composition is as follows:

[0083] (5) Homogenization: Preheat the secondary batching mixture to 60 °C, and then perform two-stage stepped homogenization; in the two-stage stepped homogenization, the pressure of the first stage homogenization is 30 MPa, and the pressure of the second stage homogenization is 5 MPa.

[0084] Mouse experiment

[0085] Use the fermentation compositions obtained in Examples 1-20 and Comparative Examples 1-5 of the present application to conduct a feeding test on mice infected with Helicobacter pylori. The specific experiment is as follows:

[0086] (1) Experimental subjects: Intragastrically administer 140 6-week-old male SPF-grade Balb / c mice with Helicobacter pylori (0.5 ml / d / rat), once a day, continuously for 2 weeks; one week after the end of intragastric administration, select 130 mice with successful modeling (mice infected with Helicobacter pylori), and randomly divide them into 26 groups, with 5 mice in each group.

[0087] (2) Experimental method: Mice in groups 1 - 20 were respectively given the fermented compositions provided in Examples 1 - 20 (0.5 ml / d / mouse); mice in groups 21 - 25 were respectively given the fermented compositions provided in Comparative Examples 1 - 5 (0.5 ml / d / mouse); mice in group 26 were given normal saline (0.5 ml / d / mouse); after continuous administration for 2 weeks, the mice were sacrificed.

[0088] (3) Experimental detection: The levels of inflammatory factors IL-6 and IL-8 in the above mice were detected, and the average value of the detection results of each group was taken, as shown in Table 4 below. The initial levels of inflammatory factors IL-6 and IL-8 in mice before modeling were 18.2 pg / mL and 31.5 pg / mL respectively; the initial levels of inflammatory factors IL-6 and IL-8 in successfully modeled mice were 83.7 pg / mL and 268.4 pg / mL respectively.

[0089] Table 4 Detection results of inflammatory factor levels and relative expression levels of related factors

[0090]

[0091] According to the detection results of Examples 1 - 20, Comparative Examples 1 - 5 and the normal saline group in Table 4, it can be seen that the inflammatory factor levels of Comparative Examples 1 - 5 and the normal saline group are significantly higher than those of Examples 1 - 20, indicating that the fermented compositions obtained in Examples 1 - 20 of the present application have a good inhibitory effect on Helicobacter pylori.

[0092] According to the detection results of Examples 1 - 7 and Comparative Examples 1 - 3, it can be known that the fermented compositions of Examples 1 - 7 of the present application can reduce the levels of inflammatory growth factors IL-6 and IL-8 to below 38 pg / mL and below 85 pg / mL respectively; while the fermented compositions of Comparative Examples 1 - 3 can only reduce the inflammatory growth factor IL-6 to 50.8 - 71.4 pg / mL and the inflammatory growth factor IL-8 to 165.7 - 202.8 pg / mL. Therefore, it shows that the present application uses Bifidobacterium longum, Saccharopolyspora, longan seed and small molecule peptides, and controls the weight parts of each within the following ranges: 5 - 15 parts of Bifidobacterium longum, 1 - 5 parts of Saccharopolyspora, 5 - 15 parts of longan seed powder and 2 - 6 parts of small molecule peptides, and a fermented composition with excellent inhibitory effect on Helicobacter pylori can be obtained.

[0093] According to the detection results of Examples 8 - 13, it can be known that the present application further performs microwave treatment on the longan seed powder, which can improve the inhibitory effect of the fermented composition on Helicobacter pylori, and controls the irradiation power of the longan seed powder within the range of 600 - 800 W and the irradiation time within the range of 15 - 25 min, and the obtained fermented composition has a better inhibitory effect on Helicobacter pylori.

[0094] According to the detection results of Example 8 and Examples 14-17, the levels of inflammatory growth factors IL-6 and IL-8 in Examples 14-16 are significantly lower than those in Example 8 and Example 17, indicating that when the weight ratio of the probiotic composition to the skim milk powder in this application is controlled within the range of (1.8-2.5):100, the obtained fermented composition has a better taste and a better consistent effect on Helicobacter pylori.

[0095] According to the detection results of Example 15, Examples 18-20, and Comparative Examples 4-5, the levels of inflammatory growth factors IL-6 and IL-8 in Example 15 and Examples 18-20 are significantly lower than those in Comparative Examples 4-5, indicating that in the preparation steps of the fermented composition of this application, when the pressure of the first-stage homogenization is controlled at 20-25 MPa and the pressure of the second-stage homogenization is controlled at 5-10 MPa, a fermented composition with a good effect of inhibiting Helicobacter pylori, delicate taste, and strong fermentation flavor can be obtained.

[0096] Although the present invention has been described in detail above with general descriptions and specific embodiments, based on the present invention, some modifications or improvements can be made, which are obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection required by the present invention.

Claims

1. A probiotic composition, characterized in that, It consists of the following components in parts by weight: 5 - 15 parts of Bifidobacterium longum, 1 - 5 parts of Saccharopolyspora, 5 - 15 parts of longan seed powder, and 2 - 6 parts of small molecule peptides; The preparation method of the longan seed powder is as follows: Wash and dry the longan seeds, scrape off the outer black skin, and grind them into powder; then place the powder in a microwave device and irradiate it at 600 - 800W for 15 - 25 minutes to obtain the longan seed powder.

2. The probiotic composition according to claim 1, wherein The probiotic composition consists of the following components in parts by weight: 5 - 15 parts of Bifidobacterium longum, 2 - 4 parts of Saccharopolyspora, 8 - 13 parts of longan seed powder, and 2 - 6 parts of small molecule peptides.

3. Use of the probiotic composition according to any one of claims 1 - 2 in the preparation of pharmaceutical adjuvants.

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