Stomach nourishing and protecting composition containing compound probiotics and application of stomach nourishing and protecting composition

Abstract

The invention provides a stomach nourishing and protecting composition containing compound probiotics and application of the stomach nourishing and protecting composition, and belongs to the technical field of nutrition and health care compositions. The stomach nourishing and protecting composition provided by the invention comprises fructo-oligosaccharide, inulin, lactitol, fermented hericium erinaceus powder, orange juice powder, konjaku flour, stachyose, tremella polysaccharide, seaweed meal, fucoidin and compound bacterial powder. The compound bacterium powder is prepared by combining bacterium powder of lactobacillus acidophilus NCFM, bacterium powder of bifidobacterium animalis subsp. Lactis Bb-12, bacterium powder of bifidobacterium lactis BR001, bacterium powder of bifidobacterium bifidum BL002, bacterium powder of bifidobacterium longum BC012, bacterium powder of lactobacillus reuteri LL029, bacterium powder of lactobacillus paracasei GF045, bacterium powder of lactobacillus rhamnosus GG, bacterium powder of lactobacillus fermentum LF028, bacterium powder of lactobacillus plantarum LZ010 and bacterium powder of lactobacillus plantarum LZ026. The composition provided by the invention has remarkable stomach nourishing and protecting effects, can effectively improve gastric mucosal injury, and has a good market prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of nutritional health-care compositions, and in particular relates to a stomach-nourishing and stomach-protecting composition containing composite probiotics and an application thereof. Background Art

[0002] The stomach is the digestive organ of the human body, located below the diaphragm, connected to the esophagus above and the small intestine below. The stomach is the main organ of the human body's digestion and absorption function. All kinds of food need to be ground by the stomach wall (gastric mucosa) and react with the gastric juice secreted by the gastric mucosa to become a paste-like chyme, and then enter the small intestine for further digestion and absorption to become nutrients. The gastric juice secreted by the gastric mucosa mainly includes gastric acid (hydrochloric acid), pepsin, gastric mucus, etc. Gastric acid can denature the protein in the food to facilitate the degradation of pepsin, and gastric acid can also activate pepsinogen into pepsin, so that it has the activity of decomposing protein. Traditional Chinese medicine believes that the spleen and stomach are the foundation of acquired health. If you want a healthy stomach, repairing the gastric mucosa is the key. Therefore, it is very meaningful to develop a composition formula with the function of nourishing and protecting the stomach. Summary of the invention

[0003] In order to solve the problems existing in the prior art, the purpose of the present invention is to provide a stomach nourishing and protecting composition, which contains 11 kinds of probiotic powders, has significant stomach nourishing and protecting effects, and can effectively improve gastric mucosal damage.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a stomach nourishing and protecting composition, which comprises the following components by weight: 35-50 parts of oligofructose, 20-25 parts of inulin, 15-25 parts of lactitol, 5-10 parts of fermented hericium erinaceus powder, 2-6 parts of orange juice powder, 2-4 parts of konjac flour, 0.5-1.5 parts of stachyose, 0.5-1.5 parts of tremella polysaccharide, 0.5-1.5 parts of seaweed powder, 2-3.5 parts of fucoidan and 2-4 parts of compound bacterial powder; The composite bacterial powder is obtained by mixing Lactobacillus acidophilus NCFM bacterial powder, Bifidobacterium animalis lactis subspecies Bb-12 bacterial powder, Bifidobacterium lactis BR001 bacterial powder, Bifidobacterium bifidum BL002 bacterial powder, Bifidobacterium longum BC012 bacterial powder, Lactobacillus reuteri LL029 bacterial powder, Lactobacillus paracasei GF045 bacterial powder, Lactobacillus rhamnosus GG bacterial powder, Lactobacillus fermentum LF028 bacterial powder, Lactobacillus plantarum LZ010 bacterial powder and Lactobacillus plantarum LZ026 bacterial powder.

[0005] Lactobacillus acidophilus NCFM powder, Bifidobacterium animalis lactis subsp. lactis Bb-12 powder, Bifidobacterium lactis BR001 powder, Bifidobacterium bifidum BL002 powder, Bifidobacterium longum BC012 powder, Lactobacillus reuteri LL029 powder, Lactobacillus paracasei GF045 powder, Lactobacillus rhamnosus GG powder, Lactobacillus fermentum LF028 powder, Lactobacillus plantarum LZ010 powder and Lactobacillus plantarum LZ026 powder are mixed according to the number of viable cells (1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3).

[0006] Preferably, the method for preparing bacterial powder comprises the following steps: inoculating the bacterial strain into a culture medium containing cactus powder for fermentation, mixing the bacterial bodies obtained by fermentation with a freeze-drying protective agent, and preparing freeze-dried powder.

[0007] Preferably, the culture medium containing cactus powder comprises: 8-12 g / L peptone, 5-15 g / L beef extract powder, 3-7 g / L yeast extract powder, 15-25 g / L glucose, 3-6 g / L sodium acetate, 1.5-2.5 g / L dipotassium hydrogen phosphate, 1.5-2.5 g / L ammonium citrate, 0.05-0.15 g / L magnesium sulfate, 0.01-0.09 g / L manganese sulfate, 0.5-1.5 g / L Tween 80, 0.2-0.8 g / L L-cysteine ​​hydrochloride and 8-12 g / L cactus powder.

[0008] Preferably, the lyoprotectant comprises 8-12 wt % skim milk powder, 5-15 wt % cactus powder, 6-10 wt % trehalose, 0.3-0.7 wt % vitamin E and 1.5-2.5 wt % lactose.

[0009] Preferably, the mass ratio of the bacterial cells to the lyophilization protective agent is 1:(1-3).

[0010] Preferably, the method for preparing the fermented Hericium erinaceus powder comprises the following steps: inoculating Aspergillus oryzae into a fermentation medium containing Hericium erinaceus powder, fermenting, sterilizing the fermentation product, and spray drying to obtain the fermented Hericium erinaceus powder.

[0011] Preferably, the fermentation medium containing Hericium erinaceus powder comprises: 150-250 g / L potato flour, 150-220 g / L Hericium erinaceus powder and 15-25 g / L glucose.

[0012] Preferably, the fermentation temperature is 30-33° C., the rotation speed is 150-200 r / min, the ventilation volume is 4.0-4.8 L / min, and the fermentation time is 38-42 h.

[0013] The present invention also provides application of the stomach nourishing and stomach protecting composition in preparing stomach nourishing and stomach protecting products.

[0014] Compared with the prior art, the technical solution of the present invention has the following beneficial effects: The invention prepares freeze-dried powders by fermenting a culture medium containing cactus powder with Lactobacillus acidophilus NCFM, Bifidobacterium animalis lactis subspecies Bb-12, Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus rhamnosus GG, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010 and Lactobacillus plantarum LZ026 respectively; the 11 freeze-dried powders are mixed to obtain composite bacterial powder; the composite bacterial powder is proportioned with fermented Hericium erinaceus powder and other components to obtain a stomach nourishing and stomach protecting composition containing composite probiotics; the composition has significant stomach nourishing and stomach protecting effects, can effectively improve gastric mucosal damage and has good market prospects. DETAILED DESCRIPTION

[0015] The invention provides a stomach nourishing and protecting composition, which comprises the following components in parts by mass: 35-50 parts of oligofructose, 20-25 parts of inulin, 15-25 parts of lactitol, 5-10 parts of fermented hericium erinaceus powder, 2-6 parts of orange juice powder, 2-4 parts of konjac flour, 0.5-1.5 parts of stachyose, 0.5-1.5 parts of tremella polysaccharide, 0.5-1.5 parts of seaweed powder, 2-3.5 parts of fucoidan and 2-4 parts of composite bacterial powder. Preferably, the ingredients include 35.8-40 parts of oligofructose, 21-24 parts of inulin, 17-20 parts of lactitol, 8-10 parts of fermented Hericium erinaceus powder, 3-4 parts of orange juice powder, 3-3.5 parts of konjac flour, 0.9-1.2 parts of stachyose, 0.8-1.2 parts of tremella polysaccharide, 0.8-1.2 parts of seaweed powder, 2.5-3.2 parts of fucoidan, and 3.1-3.5 parts of composite bacterial powder.

[0016] The composite bacterial powder of the present invention is obtained by mixing Lactobacillus acidophilus NCFM powder, Bifidobacterium animalis lactis subspecies Bb-12 powder, Bifidobacterium lactis BR001 powder, Bifidobacterium bifidum BL002 powder, Bifidobacterium longum BC012 powder, Lactobacillus reuteri LL029 powder, Lactobacillus paracasei GF045 powder, Lactobacillus rhamnosus GG powder, Lactobacillus fermentum LF028 powder, Lactobacillus plantarum LZ010 powder and Lactobacillus plantarum LZ026 powder. The number of viable bacteria in the composite bacterial powder of the present invention is 1×10 11 cfu / g~9×10 11 The composite bacterial powder of the present invention is preferably obtained by mixing the bacterial powders according to the number of viable bacteria (1-3): (1-3): (1-3): (1-3): (1-3): (1-3): (1-3): (1-3): (1-3): (1-3): (1-3), more preferably, the bacterial powders are mixed in equal proportions according to the number of viable bacteria, and the bacterial powder is preferably a strain freeze-dried powder.

[0017] The deposit number of the Bifidobacterium lactis BR001 of the present invention is CGMCC No. 23665; the deposit number of the Bifidobacterium bifidum BL002 is CGMCC No. 23664; the deposit number of the Bifidobacterium longum BC012 is CGMCC No. 23663; the deposit number of the Lactobacillus reuteri LL029 is CGMCC No. 23648; the deposit number of the Lactobacillus paracasei GF045 is CGMCC No. 26482; the deposit number of the Lactobacillus fermentum LF028 is CGMCC No. 22833; the deposit number of the Lactobacillus plantarum LZ010 is CGMCC No. 24258; the deposit number of the Lactobacillus plantarum LZ026 is CGMCC No. 22832. The Lactobacillus acidophilus NCFM was purchased from Danisco, Denmark, the Bifidobacterium animalis subspecies lactis Bb-12 was purchased from Chr. Hansen, Denmark, and the Lactobacillus rhamnosus GG was purchased from Chr. Hansen, Denmark.

[0018] In the present invention, the preparation method of bacterial powder comprises the following steps: the bacterial strain is inoculated into a culture medium containing cactus powder for fermentation, and the bacterial bodies obtained by fermentation are mixed with a freeze-drying protective agent to prepare freeze-dried powder.

[0019] Preferably, the method for preparing Lactobacillus acidophilus NCFM powder comprises the following steps: inoculating Lactobacillus acidophilus NCFM into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing animal Bifidobacterium lactis subspecies Bb-12 bacterial powder comprises the following steps: inoculating animal Bifidobacterium lactis subspecies Bb-12 into a culture medium containing cactus powder for fermentation, mixing the fermented bacterial bodies with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the preparation method of Bifidobacterium lactis BR001 powder comprises the following steps: inoculating Bifidobacterium lactis BR001 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing the powder of Bifidobacterium bifidum BL002 comprises the following steps: inoculating Bifidobacterium bifidum BL002 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the preparation method of Bifidobacterium longum BC012 powder comprises the following steps: inoculating Bifidobacterium longum BC012 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing Lactobacillus reuteri LL029 powder comprises the following steps: inoculating Lactobacillus reuteri LL029 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the preparation method of Lactobacillus paracasei GF045 bacterial powder comprises the following steps: inoculating Lactobacillus paracasei GF045 into a culture medium containing cactus powder for fermentation, mixing the fermented bacterial bodies with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing the Lactobacillus rhamnosus GG powder comprises the following steps: inoculating Lactobacillus rhamnosus GG into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing the powder of Lactobacillus fermentum LF028 comprises the following steps: inoculating Lactobacillus fermentum LF028 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing Lactobacillus plantarum LZ010 powder comprises the following steps: inoculating Lactobacillus plantarum LZ010 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder; Preferably, the method for preparing Lactobacillus plantarum LZ026 powder comprises the following steps: inoculating Lactobacillus plantarum LZ026 into a culture medium containing cactus powder for fermentation, mixing the fermented bacteria with a freeze-drying protective agent to prepare freeze-dried powder.

[0020] In the present invention, the Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus GG are preferably activated and expanded using MRS culture medium, so that each strain is in an optimal state of vitality to prepare seed liquid; the Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, and Bifidobacterium animalis subspecies lactis Bb-12 are preferably activated and expanded using improved MRS culture medium, so that each strain is in an optimal state of vitality to prepare seed liquid.

[0021] The invention carries out fermentation culture on the prepared seed liquid, wherein the culture medium containing cactus powder comprises: 8-12 g / L of peptone, 5-15 g / L of beef extract powder, 3-7 g / L of yeast extract powder, 15-25 g / L of glucose, 3-6 g / L of sodium acetate, 1.5-2.5 g / L of dipotassium hydrogen phosphate, 1.5-2.5 g / L of ammonium citrate, 0.05-0.15 g / L of magnesium sulfate, 0.01-0.09 g / L of manganese sulfate, 0.5-1.5 g / L of Tween 80, 0.2-0.8 g / L of L-cysteine ​​hydrochloride and 8-12 g / L of cactus powder. Preferably, it includes 10g / L peptone, 10g / L beef extract powder, 5g / L yeast extract powder, 20g / L glucose, 5g / L sodium acetate, 2g / L potassium dihydrogen phosphate, 2g / L ammonium citrate, 0.1g / L magnesium sulfate, 0.05g / L manganese sulfate, 1g / L Tween 80, 0.5g / L L-cysteine ​​hydrochloride and 10g / L cactus powder. The preparation method of the culture medium containing cactus powder includes: mixing the components of the culture medium, adding purified water, heating and boiling until completely dissolved, adjusting the pH to 6.9-7.1, and high temperature sterilization. The high temperature sterilization is preferably 121°C for 30min.

[0022] The conditions for the expanded culture and fermentation culture of the present invention are static culture at 36-38°C for 16-20h, preferably static culture at 37°C for 18h. After the fermentation of the present invention, centrifugal enrichment is performed to obtain the bacterial cells of each strain, and the centrifugal conditions are preferably centrifuged at 8000rmp for 20min at 4°C.

[0023] The freeze-dried protective agent of the present invention comprises 8-12wt% skim milk powder, 5-15wt% cactus powder, 6-10wt% trehalose, 0.3-0.7wt% vitamin E and 1.5-2.5wt% lactose, and the freeze-dried protective agent is supplemented with water to 100%. As an optional embodiment, the preparation method of the freeze-dried protective agent of the present invention comprises: dissolving skim milk powder, cactus powder, trehalose, vitamin E and lactose in water according to mass percentage, and obtaining the freeze-dried protective agent after high-temperature sterilization. Preferably, the mass ratio of the bacteria to the freeze-dried protective agent of the present invention is 1: (1-3), preferably 1: 2. In the present invention, the bacteria and the freeze-dried protective agent are mixed and then freeze-dried, and the freeze-drying conditions are preferably a cold trap temperature of -75°C and freeze-drying for 72h. The number of viable bacteria in each bacterial powder obtained by the present invention ranges from 1×10 11 cfu / g~9×10 11 cfu / g.

[0024] The preparation method of the fermented Hericium erinaceus powder of the present invention comprises the following steps: inoculating Aspergillus oryzae into a fermentation medium containing Hericium erinaceus powder, fermenting, sterilizing the fermentation product, spray drying, and obtaining the fermented Hericium erinaceus powder. The Aspergillus oryzae of the present invention is preferably activated and cultured using a PDA medium, expanded and cultured using a PDB medium, and seed liquid is prepared; the activation culture condition is preferably cultured on a shaking table at 30-34°C for 35-37h.

[0025] The fermentation medium containing Hericium erinaceus powder of the present invention comprises: 150-250 g / L potato flour, 150-220 g / L Hericium erinaceus powder and 15-25 g / L glucose, preferably 200 g / L potato flour, 200 g / L Hericium erinaceus powder and 20 g / L glucose. The preparation method of the culture medium comprises: mixing the components of the culture medium, adding purified water, heating and boiling until completely dissolved, adjusting the pH to 5.4-5.6, and sterilizing at high temperature. The high temperature sterilization is preferably 121°C, and the sterilization is 30 minutes. The fermentation temperature of the present invention is 30-33°C, preferably 32°C, the rotation speed is 150-200 r / min, preferably 180 r / min, the ventilation volume is 4.0-4.8 L / min, preferably 4.5 L / min, and the time is 38-42 hours, preferably 40 hours.

[0026] The present invention sterilizes the fermentation product, removes Aspergillus oryzae, preferably uses a filtering device to remove Aspergillus oryzae, and spray-dries the fermentation liquid. The spray-drying conditions are preferably: an inlet temperature of 190°C and an outlet temperature of 85°C, to obtain the spray-dried fermented Hericium erinaceus powder.

[0027] The present invention also provides the use of the above-mentioned stomach nourishing and stomach protecting composition in the preparation of stomach nourishing and stomach protecting products, and the product is preferably a medicine or a health product.

[0028] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] In a specific embodiment of the present invention: Hericium erinaceus powder and cactus powder were purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0030] Medium preparation: (1) PDA medium: 200 g / L potato flour, 20 g / L glucose, and 20 g / L agar. Mix the components of the medium, add purified water, heat and boil until completely dissolved, and sterilize at 121°C for 30 min to prepare the PDA medium.

[0031] (2) PDB medium (basic fermentation medium): 200 g / L potato flour, 20 g / L glucose. Mix the components of the medium, add purified water, heat and boil until completely dissolved, adjust the pH to 5.5, sterilize at 121°C for 30 min, and prepare the PDB medium.

[0032] (3) PDC medium (Hericium erinaceus fermentation medium): 200 g / L potato flour, 200 g / L Hericium erinaceus powder, and 20 g / L glucose. Mix the components of the medium, add purified water, heat and boil until completely dissolved, adjust the pH to 5.5, sterilize at 121°C for 30 min, and prepare the PDC medium.

[0033] (4) XRZ medium: peptone 10 g / L, beef extract powder 10 g / L, yeast extract powder 5 g / L, glucose 20 g / L, sodium acetate 5 g / L, potassium dihydrogen phosphate 2 g / L, ammonium citrate 2 g / L, magnesium sulfate 0.1 g / L, manganese sulfate 0.05 g / L, Tween 80 1 g / L, L-cysteine ​​hydrochloride 0.5 g / L, cactus powder 10 g / L; mix the components of the medium, add purified water, heat and boil until completely dissolved, adjust the pH to 7.0, sterilize at 121°C for 30 min, and prepare XRZ medium.

[0034] (5) MRS medium: MRS medium was purchased from Beijing Luqiao Technology Co., Ltd. 55.2 g of MRS medium raw material was weighed into 1 L of distilled water, heated and boiled until completely dissolved, and sterilized by high pressure at 121°C for 15 min to prepare MRS medium.

[0035] (6) Improved MRS medium: MRS medium was purchased from Beijing Luqiao Technology Co., Ltd. 55.2 g of MRS medium raw material and 0.5 g of L-cysteine ​​hydrochloride were weighed into 1 L of distilled water, heated and boiled until completely dissolved, and sterilized by high pressure at 121°C for 15 min to prepare improved MRS medium.

[0036] (7) HTG medium: peptone 10 g / L, beef extract powder 10 g / L, yeast extract powder 5 g / L, glucose 20 g / L, sodium acetate 5 g / L, potassium dihydrogen phosphate 2 g / L, ammonium citrate 2 g / L, magnesium sulfate 0.1 g / L, manganese sulfate 0.05 g / L, Tween 80 1 g / L, L-cysteine ​​hydrochloride 0.5 g / L, Hericium erinaceus powder 10 g / L; mix the components of the medium, add purified water, heat and boil until completely dissolved, adjust the pH to 7.0, sterilize at 121°C for 30 min, and prepare HTG medium.

[0037] (8) HTG-Q control medium: peptone 10 g / L, beef extract powder 10 g / L, yeast extract powder 5 g / L, glucose 20 g / L, sodium acetate 5 g / L, potassium dihydrogen phosphate 2 g / L, ammonium citrate 2 g / L, magnesium sulfate 0.1 g / L, manganese sulfate 0.05 g / L, Tween 80 1 g / L, L-cysteine ​​hydrochloride 0.5 g / L; mix the components of the medium, add purified water, heat and boil until completely dissolved, adjust the pH to 7.0, sterilize at 121°C for 30 min, and prepare the HTG-Q control medium.

[0038] The strains used in the specific embodiment of the present invention are: Aspergillus oryzae purchased from Shandong Jining Xiangyuan Biotechnology Co., Ltd.; Lactobacillus acidophilus LS001, with a preservation number of CGMCC No.23649; Bifidobacterium lactis BR001, with a preservation number of CGMCC No.23665; Bifidobacterium bifidum BL002, with a preservation number of CGMCC No.23664; Bifidobacterium longum BC012, with a preservation number of CGMCC No.23663; Lactobacillus reuteri LL029, with a preservation number of CGMCC No.23648; Lactobacillus paracasei GF045, with a preservation number of CGMCC No.26482; Lactobacillus paracasei GF027, with a preservation number of CGMCC No.22831; Lactobacillus fermentum LF028, with a preservation number of CGMCC No.22833; Lactobacillus plantarum LZ010, with a preservation number of CGMCCNo.24258; Lactobacillus plantarum LZ026, with a preservation number of CGMCC No.22832; Lactobacillus acidophilus NCFM (100 billion / g), purchased from Danisco, Denmark; Bifidobacterium animalis subsp. lactis Bb-12 (100 billion / g), purchased from Chr. Hansen, Denmark; Lactobacillus rhamnosus GG (100 billion / g), purchased from Chr. Hansen, Denmark.

[0039] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0040] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0041] Example 1 1. Explore the content of polysaccharides and β-glucan in the fermentation liquid of Hericium erinaceus powder extracted at different fermentation times: (1) Test tube slant activation culture: Aspergillus oryzae was inoculated into PDA slant culture medium and cultured at 30°C for 6 days.

[0042] (2) Preparation of spore suspension: Add 5 mL of sterile water to the slant of the activated strain, gently scrape it with an inoculating loop, pour it into a sterilized triangular flask filled with glass beads, and use the glass beads to fully disperse the spores to make 1×10 7Prepare a spore suspension of 100 / mL for later use.

[0043] (3) Seed shake flask culture: Inoculate 4 mL of spore suspension into a 250 mL Erlenmeyer flask containing 50 mL of PDB medium and culture in a constant temperature shaker at 32°C and 180 rpm for 36 h to obtain seed solution.

[0044] (4) Liquid shake flask culture: Add 1% (v / v) inoculum of Aspergillus oryzae seed liquid into 10 250 mL Erlenmeyer flasks containing 50 mL PDC medium, place in a constant temperature shaker at 32°C and 180 r / min, and take one flask for testing at 0 h, 8 h, 16 h, 24 h, 32 h, 40 h, 48 h, 56 h, 64 h, and 72 h. Repeat this experiment three times.

[0045] (5) β-glucan content detection: The Megazyme kit (K-YBGL) was used for determination. The β-glucan yield was calculated according to formula (1): Formula (1).

[0046] (6) Polysaccharide extraction: Take a certain amount of fermentation liquid and centrifuge (4000g, 30min), add anhydrous ethanol to the supernatant (the volume ratio of supernatant to anhydrous ethanol is 25:75) for alcohol precipitation, let it stand for 24h and then centrifuge (4200g, 30min), and keep the precipitate; add water to the precipitate at 65℃ and freeze-dry to obtain Hericium erinaceus extract. The polysaccharide content in the Hericium erinaceus extract obtained by extraction was determined by the phenol-sulfuric acid method, and the polysaccharide yield was calculated by formula (2): Formula (2).

[0047] The polysaccharide and β-glucan contents extracted at different fermentation times are shown in Table 1. The “*” in the table indicates that compared with 0h, P <0.05.

[0048] Table 1 Polysaccharide and β-glucan content in each group

[0049] The results showed that during the fermentation process of Aspergillus oryzae, as the fermentation time prolonged, the content of Hericium erinaceus polysaccharide and β-glucan gradually increased, reaching the highest content at 40h, and the content of Hericium erinaceus polysaccharide and β-glucan decreased from 48h to 72h. The possible reasons for the change in the content of Hericium erinaceus polysaccharide and β-glucan are: the various enzymes secreted by Aspergillus oryzae, such as amylase, saccharifying enzyme, cellulase, phytase, pectinase, etc., enzymatically hydrolyze Hericium erinaceus in the early stage of fermentation, which increases the content of Hericium erinaceus polysaccharide and β-glucan. In the late stage of fermentation, as Aspergillus oryzae grows, it decomposes and consumes Hericium erinaceus polysaccharide and β-glucan.

[0050] 2. Explore the differences between fermented and unfermented Hericium erinaceus powder (1) Preparation of fermented Hericium erinaceus powder: Aspergillus oryzae seed liquid was prepared by the same method as step 1, and the Aspergillus oryzae seed liquid was inoculated into a fermentation tank containing 5 L PDC medium at an inoculum rate of 1% (v / v) at 32°C, 180 r / min, and a ventilation rate of 4.5 L / min. The fermentation was terminated after 40 h. After the Aspergillus oryzae cells were removed by a 60-mesh filter, the fermentation liquid was spray-dried at an inlet temperature of 190°C and an outlet temperature of 85°C to obtain spray-dried Hericium erinaceus powder.

[0051] (2) Preparation of unfermented Hericium erinaceus powder: The sterilized PDC culture medium was directly spray-dried with an inlet temperature of 190°C and an outlet temperature of 85°C to obtain spray-dried Hericium erinaceus powder that was not fermented with Aspergillus oryzae.

[0052] The same method as step 1 was used to determine the β-glucan and polysaccharide contents of fermented Hericium erinaceus powder and unfermented Hericium erinaceus powder, respectively. The test results are shown in Table 2. "**" in the table indicates that compared with unfermented Hericium erinaceus powder, P <0.01.

[0053] Table 2 Polysaccharide and β-glucan contents of fermented Hericium erinaceus powder and unfermented Hericium erinaceus powder

[0054] The results showed that the polysaccharide and β-glucan contents of Hericium erinaceus powder were greatly increased after fermentation by Aspergillus oryzae.

[0055] Example 2 1. Effect of cactus powder addition on lactic acid bacteria Cactus powder was used to culture lactic acid bacteria to detect whether cactus powder is beneficial to the growth of lactic acid bacteria.

[0056] MRS medium was used to culture lactobacilli: Lactobacillus acidophilus LS001, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus paracasei GF027, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus GG; the above strains were inoculated into sterilized MRS medium, cultured at 37°C for 18 h, subcultured and activated twice, and then inoculated into sterilized MRS medium again at 3% (v / v), cultured at 37°C for 18 h as seed solution for later use.

[0057] The modified MRS medium was used to culture bifidobacteria: Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, and Bifidobacterium animalis subsp. lactis Bb-12; the above strains were inoculated into sterilized modified MRS medium, cultured anaerobically at 37°C for 18 h, cultured for two consecutive times, and then inoculated into sterilized MRS medium again at 3% (v / v), cultured at 37°C for 18 h as seed solution for later use.

[0058] After the 13 strains were expanded and cultured, the seed liquid was inoculated into XRZ medium at a 3% (v / v) inoculation rate, and the pH, OD value and viable count were tested at 37°C for 16 hours under anaerobic conditions. The results are shown in Table 3.

[0059] Table 3 pH, OD values ​​and viable counts of 13 strains

[0060] The results showed that not all lactic acid bacteria could grow well in XRZ medium. Among them, Lactobacillus acidophilus LS001 and Lactobacillus paracasei GF027 grew slowly, had high pH, ​​low OD value and few viable bacteria. Cactus powder had an inhibitory effect on the growth of these two lactic acid bacteria. A total of 11 strains, including Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis Bb-12 and Lactobacillus rhamnosus GG, could grow normally in XRZ medium with a viable bacteria count of 10 9 After that, the experiment was conducted with 11 strains, and Lactobacillus acidophilus LS001 and Lactobacillus paracasei GF027 were removed.

[0061] 2. Experiment on the effect of Hericium erinaceus powder on lactic acid bacteria In this study, Hericium erinaceus powder was used to culture lactic acid bacteria to detect whether Hericium erinaceus powder is beneficial to the growth of lactic acid bacteria.

[0062] The seed liquids of Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus GG, Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, and Bifidobacterium animalis subspecies lactis Bb-12 were prepared respectively by the same method as step 1.

[0063] The seed liquid of the 11 strains was inoculated into HTG and HTG-Q medium at a 3% (v / v) inoculation rate, and the number of viable bacteria was detected after anaerobic static culture at 37°C for 16 hours. The results are shown in Table 4.

[0064] Table 4 pH, OD values ​​and viable counts of 11 strains

[0065] The results of comparing the two culture media with and without Hericium erinaceus powder showed that Hericium erinaceus powder was beneficial to the increase of the number of live lactic acid bacteria. The reason may be that Hericium erinaceus contains a variety of bioactive substances, such as polysaccharides, oligosaccharides, sterols, terpenes and phenols, adenosine, unsaturated fatty acids, proteins and peptides, which have a good growth-promoting effect on bacteria. Therefore, adding Hericium erinaceus powder is beneficial to the growth of lactic acid bacteria.

[0066] 3. Comparison between cactus powder lactic acid bacteria powder and ordinary lactic acid bacteria powder (1) Preparation of cactus powder lactic acid bacteria powder: The seed liquid preparation of Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis Bb-12, and Lactobacillus rhamnosus GG was the same as step 1. The seed liquid was inoculated into XRZ medium at 3% (v / v) at 37°C, and after anaerobic static culture for 16 hours, the strains were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C.

[0067] The bacterial cells of each strain were fully mixed with the protective agent at a mass ratio of 1:2, and freeze-dried for 72 hours at a cold trap temperature of -75°C to obtain freeze-dried powder of each strain, and the live bacteria of each freeze-dried powder were counted.

[0068] The protective agent is: skim milk powder 10wt%, cactus powder 10wt%, trehalose 8wt%, vitamin E 0.5wt%, lactose 2wt%, and the rest is water. Skim milk powder, cactus powder, trehalose, vitamin E and lactose are dissolved in water according to mass percentage, and the freeze-dried protective agent is obtained after sterilization at 110°C for 20 minutes.

[0069] (2) Preparation of ordinary lactic acid bacteria powder The seed liquid preparation of Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis Bb-12, and Lactobacillus rhamnosus GG was the same as step 1. The seed liquid was inoculated into MRS medium at 3% (v / v) at 37°C, and after anaerobic static culture for 16 hours, the strains were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C.

[0070] The bacterial cells of each strain were fully mixed with the protective agent at a mass ratio of 1:2, and freeze-dried for 72 hours at a cold trap temperature of -75°C to obtain freeze-dried powder of each strain, and the live bacteria of each freeze-dried powder were counted.

[0071] The protective agent is: skim milk powder 10wt%, maltodextrin 10wt%, trehalose 8wt%, vitamin E 0.5wt%, lactose 2wt%, and the rest is water. Skim milk powder, maltodextrin, trehalose, vitamin E and lactose are dissolved in water according to mass percentage, and the freeze-dried protective agent is obtained after sterilization at 110°C for 20 minutes.

[0072] The viable bacteria counts of cactus powder lactic acid bacteria powder and ordinary lactic acid bacteria powder were compared, and the results are shown in Table 5.

[0073] Table 5 The number of viable bacteria in different bacterial powders prepared from 11 strains

[0074] The results showed that the number of live bacteria in the freeze-dried powder of the 11 strains in both culture and freeze-drying methods was within 10 11 The above results showed no difference, and cactus powder had no adverse effect on the strains.

[0075] Example 3 A stomach nourishing and protecting composition is prepared by mixing 50g of oligofructose, 20g of inulin, 15g of lactitol, 5g of fermented hericium erinaceus powder, 2g of orange juice powder, 2g of konjac powder, 0.5g of stachyose, 0.5g of tremella polysaccharide, 0.5g of seaweed powder, 2.5g of fucoidan and 2g of composite bacterial powder.

[0076] Preparation of fermented Hericium erinaceus powder: (1) Test tube slant activation culture: Aspergillus oryzae was inoculated into PDA slant culture medium and cultured at 30°C for 6 days.

[0077] (2) Preparation of spore suspension: Add 5 mL of sterile water to the slant of the activated strain, gently scrape it with an inoculating loop, pour it into a sterilized triangular flask filled with glass beads, and use the glass beads to fully disperse the spores to make 1×10 7 Prepare a spore suspension of 100 / mL for later use.

[0078] (3) Seed shake flask culture: inoculate 4 mL of spore suspension into a 250 mL Erlenmeyer flask containing 50 mL of PDB medium and culture in a constant temperature shaker at 32°C and 180 rpm for 36 h.

[0079] (4) The Aspergillus oryzae seed liquid was inoculated into a fermentation tank containing 5 L PDC medium at a 1% inoculum rate at 32 °C, 180 r / min, and a ventilation volume of 4.5 L / min. The fermentation was terminated after 40 h. The Aspergillus oryzae fungi were removed by a 60-mesh filter. The fermentation liquid was spray-dried at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain spray-dried Hericium erinaceus powder.

[0080] Preparation of composite bacterial powder: Seed solutions were prepared according to step 1 of Example 2 for Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis Bb-12, and Lactobacillus rhamnosus GG.

[0081] (1) Preparation of Bifidobacterium lactis BR001 freeze-dried powder: The seed solution of Bifidobacterium lactis BR001 was inoculated into XRZ medium at 3% (v / v) at 37°C, and cultured anaerobically for 16 hours. The cells were then centrifuged at 8000 rpm for 20 minutes at 4°C to obtain enriched cells. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0082] (2) Preparation of freeze-dried powder of Bifidobacterium bifidum BL002: The seed liquid of Bifidobacterium bifidum BL002 was inoculated into XRZ medium at 37°C at a rate of 3% (v / v), and cultured anaerobically for 16 hours. The cells were then centrifuged at 8000 rpm for 20 minutes at 4°C to obtain the enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0083] (3) Preparation of Bifidobacterium longum BC012 freeze-dried powder: The seed liquid of Bifidobacterium longum BC012 was inoculated into XRZ medium at 37°C at a rate of 3% (v / v), and cultured anaerobically for 16 hours. The cells were then centrifuged at 8000 rpm for 20 minutes at 4°C to obtain enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0084] (4) Preparation of freeze-dried powder of Lactobacillus reuteri LL029: The seed liquid of Lactobacillus reuteri LL029 was inoculated into XRZ medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 hours, the bacteria were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C. The bacteria and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried for 72 hours at a cold trap temperature of -75°C to obtain freeze-dried powder.

[0085] (5) Preparation of freeze-dried powder of Lactobacillus paracasei GF045: The seed liquid of Lactobacillus paracasei GF045 was inoculated into XRZ medium at 37°C at a 3% (v / v) inoculation rate, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain the enriched cells. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain freeze-dried powder.

[0086] (6) Preparation of lyophilized powder of Lactobacillus fermentum LF028: The seed liquid of Lactobacillus fermentum LF028 was inoculated into XRZ medium at 37°C at a rate of 3% (v / v), and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain the enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the lyophilized powder.

[0087] (7) Preparation of lyophilized powder of Lactobacillus plantarum LZ010: The seed liquid of Lactobacillus plantarum LZ010 was inoculated into XRZ medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 h, the cells were enriched by centrifugation at 8000 rpm for 20 min at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried for 72 h at a cold trap temperature of -75°C to obtain lyophilized powder.

[0088] (8) Preparation of lyophilized powder of Lactobacillus plantarum LZ026: The seed liquid of Lactobacillus plantarum LZ026 was inoculated into XRZ medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 h, the cells were enriched by centrifugation at 8000 rpm for 20 min at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0089] (9) Preparation of lyophilized powder of Lactobacillus acidophilus NCFM: 3% (v / v) of Lactobacillus acidophilus NCFM seed solution was inoculated into XRZ medium at 37°C, cultured anaerobically for 16 h, and then centrifuged at 8000 rpm for 20 min at 4°C to obtain enriched bacteria. The bacteria and protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0090] (10) Preparation of freeze-dried powder of animal Bifidobacterium lactis subsp. Bb-12: The seed liquid of animal Bifidobacterium lactis subsp. Bb-12 was inoculated into XRZ medium at a rate of 3% (v / v) at 37°C, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain freeze-dried powder.

[0091] (11) Preparation of lyophilized powder of Lactobacillus rhamnosus GG: The seed liquid of Lactobacillus rhamnosus GG was inoculated into XRZ medium at a rate of 3% (v / v) at 37°C, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0092] The protective agent is: skim milk powder 10wt%, cactus powder 10wt%, trehalose 8wt%, vitamin E 0.5wt%, lactose 2wt%, and the rest is water. Skim milk powder, cactus powder, trehalose, vitamin E and lactose are dissolved in water according to mass percentage, and the freeze-dried protective agent is obtained after sterilization at 110°C for 20 minutes.

[0093] (12) The freeze-dried powder of Lactobacillus acidophilus NCFM, the freeze-dried powder of Bifidobacterium animalis subsp. lactis Bb-12, the freeze-dried powder of Bifidobacterium lactis BR001, the freeze-dried powder of Bifidobacterium bifidum BL002, the freeze-dried powder of Bifidobacterium longum BC012, the freeze-dried powder of Lactobacillus reuteri LL029, the freeze-dried powder of Lactobacillus paracasei GF045, the freeze-dried powder of Lactobacillus rhamnosus GG, the freeze-dried powder of Lactobacillus fermentum LF028, the freeze-dried powder of Lactobacillus plantarum LZ010, and the freeze-dried powder of Lactobacillus plantarum LZ026 were fully mixed according to the number of viable cells to prepare a composite bacterial powder (the number of viable cells was on the order of 10 11 cfu / g).

[0094] Example 4 A stomach nourishing and protecting composition is prepared by mixing 35g of oligofructose, 25g of inulin, 15g of lactitol, 5g of fermented hericium erinaceus powder, 6g of orange juice powder, 2g of konjac powder, 1.5g of stachyose, 1.5g of tremella polysaccharide, 1.5g of seaweed powder, 3.5g of fucoidan and 4g of composite bacterial powder.

[0095] The preparation method of fermented Hericium erinaceus powder and the preparation method of composite fungus powder are the same as those in Example 3.

[0096] Example 5 A stomach nourishing and protecting composition is prepared by mixing 35g of oligofructose, 20g of inulin, 25g of lactitol, 5g of fermented hericium erinaceus powder, 3g of orange juice powder, 4g of konjac powder, 1.0g of stachyose, 0.8g of tremella polysaccharide, 1.2g of seaweed powder, 2g of fucoidan and 3g of composite bacterial powder.

[0097] The preparation method of fermented Hericium erinaceus powder and the preparation method of composite fungus powder are the same as those in Example 3.

[0098] Example 6 A stomach nourishing and protecting composition is prepared by mixing 35.8 g of oligofructose, 21 g of inulin, 17 g of lactitol, 10 g of fermented hericium erinaceus powder, 4 g of orange juice powder, 3 g of konjac flour, 0.9 g of stachyose, 1.2 g of tremella polysaccharide, 0.8 g of seaweed powder, 3.2 g of fucoidan, and 3.1 g of composite bacterial powder.

[0099] The preparation method of fermented Hericium erinaceus powder and the preparation method of composite fungus powder are the same as those in Example 3.

[0100] Example 7 A stomach nourishing and protecting composition is prepared by mixing 40g of oligofructose, 22g of inulin, 19.5g of lactitol, 7g of fermented hericium erinaceus powder, 2g of orange juice powder, 3g of konjac powder, 0.5g of stachyose, 0.5g of tremella polysaccharide, 0.6g of seaweed powder, 2.4g of fucoidan and 2.5g of composite bacterial powder.

[0101] The preparation method of fermented Hericium erinaceus powder and the preparation method of composite fungus powder are the same as those in Example 3.

[0102] Example 8 A stomach nourishing and protecting composition is obtained by mixing 42g of oligofructose, 20g of inulin, 21g of lactitol, 6g of fermented hericium erinaceus powder, 2g of orange juice powder, 2g of konjac powder, 1.2g of stachyose, 1.1g of tremella polysaccharide, 0.7g of seaweed powder, 2g of fucoidan, and 2g of composite bacterial powder.

[0103] The preparation method of fermented Hericium erinaceus powder and the preparation method of composite fungus powder are the same as those in Example 3.

[0104] Comparative Example 1 A stomach nourishing and protecting composition is prepared by mixing 50g of oligofructose, 20g of inulin, 15g of lactitol, 5g of hericium erinaceus powder, 2g of orange juice powder, 2g of konjac powder, 0.5g of stachyose, 0.5g of tremella polysaccharide, 0.5g of seaweed powder, 2.5g of fucoidan and 2g of cactus powder.

[0105] The preparation method of Hericium erinaceus powder is as follows: the sterilized PDC culture medium is directly spray-dried, the inlet temperature is 190° C., and the outlet temperature is 85° C., so as to obtain the spray-dried Hericium erinaceus powder which is not fermented by Aspergillus oryzae.

[0106] Comparative Example 2 A stomach nourishing and protecting composition is obtained by mixing 35.8g of oligofructose, 21g of inulin, 17g of lactitol, 10g of hericium erinaceus powder, 4g of orange juice powder, 3g of konjac powder, 0.9g of stachyose, 1.2g of tremella polysaccharide, 0.8g of seaweed powder, 3.2g of fucoidan, and 3.1g of composite bacterial powder.

[0107] The preparation method of Hericium erinaceus powder is as follows: the sterilized PDC culture medium is directly spray-dried, the inlet temperature is 190° C., and the outlet temperature is 85° C., so as to obtain the spray-dried Hericium erinaceus powder which is not fermented by Aspergillus oryzae.

[0108] Preparation of composite bacterial powder: Seed solutions were prepared according to step 1 of Example 2 for Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Lactobacillus reuteri LL029, Lactobacillus paracasei GF045, Lactobacillus fermentum LF028, Lactobacillus plantarum LZ010, Lactobacillus plantarum LZ026, Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis Bb-12, and Lactobacillus rhamnosus GG.

[0109] (1) Preparation of Bifidobacterium lactis BR001 freeze-dried powder: The seed solution of Bifidobacterium lactis BR001 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 hours, the cells were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0110] (2) Preparation of freeze-dried powder of Bifidobacterium bifidum BL002: The seed liquid of Bifidobacterium bifidum BL002 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 hours, the cells were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0111] (3) Preparation of Bifidobacterium longum BC012 freeze-dried powder: The seed liquid of Bifidobacterium longum BC012 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 hours, the cells were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 hours to obtain freeze-dried powder.

[0112] (4) Preparation of freeze-dried powder of Lactobacillus reuteri LL029: The seed liquid of Lactobacillus reuteri LL029 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 hours, the bacteria were enriched by centrifugation at 8000 rpm for 20 minutes at 4°C. The bacteria and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried for 72 hours at a cold trap temperature of -75°C to obtain freeze-dried powder.

[0113] (5) Preparation of freeze-dried powder of Lactobacillus paracasei GF045: The seed liquid of Lactobacillus paracasei GF045 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain the enriched cells. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain freeze-dried powder.

[0114] (6) Preparation of lyophilized powder of Lactobacillus fermentum LF028: The seed liquid of Lactobacillus fermentum LF028 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain the enriched cells. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the lyophilized powder.

[0115] (7) Preparation of lyophilized powder of Lactobacillus plantarum LZ010: The seed liquid of Lactobacillus plantarum LZ010 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 h, the cells were enriched by centrifugation at 8000 rpm for 20 min at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0116] (8) Preparation of lyophilized powder of Lactobacillus plantarum LZ026: The seed liquid of Lactobacillus plantarum LZ026 was inoculated into MRS medium at 37°C at a 3% (v / v) inoculation rate. After anaerobic static culture for 16 h, the cells were enriched by centrifugation at 8000 rpm for 20 min at 4°C. The cells and the protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0117] (9) Preparation of lyophilized powder of Lactobacillus acidophilus NCFM: 3% (v / v) inoculation amount of Lactobacillus acidophilus NCFM seed liquid was inoculated into MRS medium at 37°C, cultured anaerobically for 16 h, and then centrifuged at 8000 rpm for 20 min at 4°C to obtain enriched bacteria. The bacteria and protective agent were fully mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain lyophilized powder.

[0118] (10) Preparation of freeze-dried powder of animal Bifidobacterium lactis subsp. Bb-12: The seed liquid of animal Bifidobacterium lactis subsp. Bb-12 was inoculated into MRS medium at a rate of 3% (v / v) at 37°C, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain freeze-dried powder.

[0119] (11) Preparation of lyophilized powder of Lactobacillus rhamnosus GG: The seed liquid of Lactobacillus rhamnosus GG was inoculated into MRS medium at a rate of 3% (v / v) at 37°C, and cultured anaerobically for 16 h. The cells were then centrifuged at 8000 rpm for 20 min at 4°C to obtain the enriched cells. The cells and the protective agent were thoroughly mixed at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the lyophilized powder.

[0120] The protective agent is: skim milk powder 10wt%, maltodextrin 10wt%, trehalose 8wt%, vitamin E 0.5wt%, lactose 2wt%, and the rest is water. Skim milk powder, maltodextrin, trehalose, vitamin E and lactose are dissolved in water according to mass percentage, and the freeze-dried protective agent is obtained after sterilization at 110°C for 20 minutes.

[0121] (12) The freeze-dried powder of Lactobacillus acidophilus NCFM, the freeze-dried powder of Bifidobacterium animalis subsp. lactis Bb-12, the freeze-dried powder of Bifidobacterium lactis BR001, the freeze-dried powder of Bifidobacterium bifidum BL002, the freeze-dried powder of Bifidobacterium longum BC012, the freeze-dried powder of Lactobacillus reuteri LL029, the freeze-dried powder of Lactobacillus paracasei GF045, the freeze-dried powder of Lactobacillus rhamnosus GG, the freeze-dried powder of Lactobacillus fermentum LF028, the freeze-dried powder of Lactobacillus plantarum LZ010, and the freeze-dried powder of Lactobacillus plantarum LZ026 were fully mixed according to the number of viable cells to prepare a composite bacterial powder (the number of viable cells was on the order of 10 11 cfu / g).

[0122] Comparative Example 3 A stomach nourishing and protecting composition is obtained by mixing 35g of oligofructose, 20g of inulin, 25g of lactitol, 5g of hericium erinaceus powder, 3g of orange juice powder, 4g of konjac powder, 1.0g of stachyose, 0.8g of tremella polysaccharide, 1.2g of seaweed powder, 2g of fucoidan and 3g of composite bacterial powder.

[0123] The preparation method of Hericium erinaceus powder is as follows: the sterilized PDC culture medium is directly spray-dried, the inlet temperature is 190° C., and the outlet temperature is 85° C., so as to obtain the spray-dried Hericium erinaceus powder which is not fermented by Aspergillus oryzae.

[0124] The preparation method of the composite bacterial powder is the same as that in Example 3.

[0125] Comparative Example 4 A stomach nourishing and protecting composition is prepared by mixing 40g of oligofructose, 22g of inulin, 19.5g of lactitol, 7g of fermented hericium erinaceus powder, 2g of orange juice powder, 3g of konjac powder, 0.5g of stachyose, 0.5g of tremella polysaccharide, 0.6g of seaweed powder, 2.4g of fucoidan and 2.5g of composite bacterial powder.

[0126] The preparation method of fermented Hericium erinaceus powder is the same as that in Example 3.

[0127] The preparation method of the composite bacterial powder is the same as that of Comparative Example 2.

[0128] Comparative Example 5 A stomach nourishing and protecting composition is obtained by mixing 42g of oligofructose, 20g of inulin, 21g of lactitol, 6g of hericium erinaceus powder, 2g of orange juice powder, 2g of konjac powder, 1.2g of stachyose, 1.1g of tremella polysaccharide, 0.7g of seaweed powder, 2g of fucoidan, and 2g of composite bacterial powder.

[0129] The preparation method of fermented Hericium erinaceus powder is the same as that in Example 3.

[0130] Composite bacterial powder: a mixture of Lactobacillus acidophilus NCFM powder, Bifidobacterium animalis lactis subspecies Bb-12 powder, Bifidobacterium bifidum BL002 powder, Bifidobacterium longum BC012 powder, Lactobacillus paracasei GF045 powder, Lactobacillus rhamnosus GG powder, Lactobacillus fermentum LF028 powder and Lactobacillus plantarum LZ010 powder (the number of viable cells is 10 11 cfu / g). The composite bacterial powder of this comparative example does not contain Bifidobacterium lactis BR001 bacterial powder, Lactobacillus reuteri LL029 bacterial powder, and Lactobacillus plantarum LZ026 bacterial powder. The preparation methods of each bacterial powder are the same as those in Example 3.

[0131] Test Example 1 Animal acute gastric mucosal injury experiment: Use the 18th item "Auxiliary protection of gastric mucosa test method" in the "Health Food Function Test and Evaluation Method (2022 Edition)".

[0132] Experimental animals: 140 healthy male Wistar rats, weighing 180-220 g.

[0133] The experiment was set as experimental groups 1 to 6, comparative groups 7 to 11, blank control group, model control group and positive control group, a total of 14 groups, 10 animals in each group, gavage once a day, gavage volume 1.0mL / 100g, for 30 consecutive days. Experimental groups 1 to 6 correspond to the compositions prepared in Examples 3, 4, 5, 6, 7 and 8, respectively, and comparative groups 7 to 11 represent the compositions prepared in Comparative Examples 1, 2, 3, 4 and 5, respectively, and the gavage dose of each group is 50mg / kg each time (dissolved in saline for gavage); the positive control group was gavaged with cimetidine, and the gavage dose of cimetidine was 0.04g / kg each time (dissolved in saline for gavage); the blank control group and the model control group were given equal volumes of saline.

[0134] After 30 days of intragastric administration in each group, all animals were strictly fasted for 24 hours (water was not allowed), and the test substance was also prohibited during this period. Except for the blank control, all experimental groups were given 1.0 mL of anhydrous ethanol per animal. The animals were killed 1 hour later, the complete stomach was exposed, the pylorus was ligated, and an appropriate amount of 10% formalin solution was perfused. After fixation for 20 minutes, the stomach was cut open along the greater curvature, the stomach contents were washed, the gastric mucosa was unfolded, and the length and width of the bleeding point or bleeding band were measured with a vernier caliper under a stereoscopic dissection microscope. Because the severity of the injury represented by the width is much greater than the length, double points are used. The scoring criteria are shown in Table 6, and the experimental results are shown in Table 7.

[0135] Table 6 Gross observation scoring criteria for acute anhydrous ethanol injury

[0136] Table 7 Gastric injury scores in each group

[0137] Note: Compared with the control group, * P <0.05;** P <0.01. Compared with the model group, # P <0.05;## P <0.01.

[0138] The results showed that compared with the control group, the rat acute gastric mucosal injury alcohol model was successfully established; compared with the model group, the compound ratios of 1 to 6 in the experimental group could significantly reduce gastric mucosal injury, and the difference was extremely significant ( P <0.01); compared with the model group, the 7-11 compound ratios in the control group can reduce gastric mucosal damage, and the difference is significant ( P<0.05); there was no significant difference between the positive control group and the experimental group; the effect of the comparison group 9 was significantly different from that of the experimental group, indicating that the effect of the Hericium erinaceus powder not treated with Aspergillus oryzae was biased; the effect of the comparison group 7 was significantly different from that of the experimental group, indicating that the Hericium erinaceus powder not treated with Aspergillus oryzae and the effect of not adding composite lactic acid bacteria were poor; the effect of the comparison group 8 was significantly different from that of the experimental group, indicating that the Hericium erinaceus powder not treated with Aspergillus oryzae and the composite bacterial powder not treated with cactus powder had poor effects; the effect of the comparison group 10 was significantly different from that of the experimental group, indicating that the composite bacterial powder was slightly worse without the embedding treatment of cactus powder; compared with the experimental group, the composite bacterial powder in the comparison group 11 lacked three kinds of powders, namely, Bifidobacterium lactis BR001 powder, Lactobacillus reuteri LL029 powder and Lactobacillus plantarum LZ026 powder, and the effect became worse, indicating that the bacterial powders had a synergistic effect with each other in the process of protecting gastric mucosal damage, which could effectively reduce gastric mucosal damage, and the effect of the present invention could not be achieved without some bacterial powders. This animal experiment shows that the combination of Hericium erinaceus fermented with Aspergillus oryzae, lactic acid bacteria fermented with cactus powder and the composite bacterial powder has a better protective effect on gastric mucosal damage.

[0139] Test Example 2 Crowd trial experiment: Subject inclusion criteria: Volunteers who meet the diagnostic criteria for chronic superficial gastritis and are diagnosed with gastric mucosal damage by gastroscopy screening. A total of 83 subjects, aged between 18 and 60 years old. Diagnostic criteria for chronic superficial gastritis: Protracted course of disease, with varying degrees of clinical symptoms such as indigestion, upper abdominal pain, heartburn, belching, acid reflux, abdominal distension, and mild tenderness in the upper abdomen. Meet the diagnostic criteria for chronic superficial gastritis by fiber gastroscopy and biopsy. Patients with gastric ulcers were excluded.

[0140] This study used the formula of Example 6, 4 g per pack. The subjects filled out a questionnaire before consumption, consumed 2 packs per day, either directly or with warm water below 40°C, for 30 consecutive days, and then filled out a questionnaire again to determine the improvement of the clinical manifestations of gastric mucosal injury.

[0141] Clinical symptoms include stomachache, belching, acid reflux, abdominal distension, loss of appetite, and less food intake. Physical signs include the degree of tenderness under the xiphoid process. The score is calculated according to the severity of the symptoms (3 points for severe cases, 2 points for moderate cases, and 1 point for mild cases), see Table 8. The experiment was terminated after 30 days of consumption, and the questionnaire results are shown in Table 9.

[0142] Table 8 Symptom severity classification table for human food trial

[0143] Table 9 Questionnaire results after 30 days of consumption

[0144] According to the questionnaire answers of the subjects, the composition formula of the present invention can effectively improve the stomach discomfort of the subjects: the subjects with acid reflux, abdominal distension and loss of appetite have been significantly improved; the subjects with belching are second; the subjects with stomach pain have the lowest improvement rate. The total improvement rate of the five indicators reached 73.28%, indicating that the formula of the present invention can bring good improvement to the subjects with gastric mucosal damage.

[0145] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A stomach nourishing and stomach protecting composition, characterized in that: By mass, it includes the following components: 35-50 parts of oligofructose, 20-25 parts of inulin, 15-25 parts of lactitol, 5-10 parts of fermented hericium erinaceus powder, 2-6 parts of orange juice powder, 2-4 parts of konjac flour, 0.5-1.5 parts of stachyose, 0.5-1.5 parts of tremella polysaccharide, 0.5-1.5 parts of seaweed powder, 2-3.5 parts of fucoidan and 2-4 parts of compound bacterial powder; The composite bacterial powder is obtained by mixing Lactobacillus acidophilus NCFM bacterial powder, Bifidobacterium animalis lactis subspecies Bb-12 bacterial powder, Bifidobacterium lactis BR001 bacterial powder, Bifidobacterium bifidum BL002 bacterial powder, Bifidobacterium longum BC012 bacterial powder, Lactobacillus reuteri LL029 bacterial powder, Lactobacillus paracasei GF045 bacterial powder, Lactobacillus rhamnosus GG bacterial powder, Lactobacillus fermentum LF028 bacterial powder, Lactobacillus plantarum LZ010 bacterial powder and Lactobacillus plantarum LZ026 bacterial powder.

2. The stomach nourishing and stomach protecting composition according to claim 1, characterized in that: Lactobacillus acidophilus NCFM powder, Bifidobacterium animalis lactis subsp. lactis Bb-12 powder, Bifidobacterium lactis BR001 powder, Bifidobacterium bifidum BL002 powder, Bifidobacterium longum BC012 powder, Lactobacillus reuteri LL029 powder, Lactobacillus paracasei GF045 powder, Lactobacillus rhamnosus GG powder, Lactobacillus fermentum LF028 powder, Lactobacillus plantarum LZ010 powder and Lactobacillus plantarum LZ026 powder are mixed according to the number of viable cells (1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3).

3. The stomach nourishing and stomach protecting composition according to claim 1, characterized in that: The preparation method of bacterial powder comprises the following steps: inoculating the bacterial strain into a culture medium containing cactus powder for fermentation, mixing the bacterial bodies obtained by fermentation with a freeze-drying protective agent, and preparing freeze-dried powder.

4. The stomach nourishing and stomach protecting composition according to claim 3, characterized in that: The culture medium containing cactus powder comprises: 8-12 g / L of peptone, 5-15 g / L of beef extract powder, 3-7 g / L of yeast extract powder, 15-25 g / L of glucose, 3-6 g / L of sodium acetate, 1.5-2.5 g / L of dipotassium hydrogen phosphate, 1.5-2.5 g / L of ammonium citrate, 0.05-0.15 g / L of magnesium sulfate, 0.01-0.09 g / L of manganese sulfate, 0.5-1.5 g / L of Tween 80, 0.2-0.8 g / L of L-cysteine ​​hydrochloride and 8-12 g / L of cactus powder.

5. The stomach nourishing and stomach protecting composition according to claim 3, characterized in that: The freeze-drying protective agent comprises 8-12 wt % of skim milk powder, 5-15 wt % of cactus powder, 6-10 wt % of trehalose, 0.3-0.7 wt % of vitamin E and 1.5-2.5 wt % of lactose.

6. The stomach nourishing and stomach protecting composition according to claim 3, characterized in that: The mass ratio of the bacteria to the freeze-dried protective agent is 1: (1-3).

7. The stomach nourishing and stomach protecting composition according to claim 1, characterized in that: The preparation method of the fermented Hericium erinaceus powder comprises the following steps: inoculating Aspergillus oryzae into a fermentation medium containing Hericium erinaceus powder, fermenting, sterilizing the fermentation product, and spray drying to obtain the fermented Hericium erinaceus powder.

8. The stomach nourishing and stomach protecting composition according to claim 7, characterized in that: The fermentation medium containing Hericium erinaceus powder comprises: 150-250 g / L potato flour, 150-220 g / L Hericium erinaceus powder and 15-25 g / L glucose.

9. The stomach nourishing and stomach protecting composition according to claim 7, characterized in that: The fermentation temperature is 30-33° C., the rotation speed is 150-200 r / min, the ventilation volume is 4.0-4.8 L / min, and the fermentation time is 38-42 h.

10. Use of the stomach nourishing and stomach protecting composition according to any one of claims 1 to 9 in the preparation of stomach nourishing and stomach protecting products.

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