A stomach-nourishing and stomach-protecting composition containing compound probiotics and its application

Abstract

The present invention provides a stomach-nourishing and protecting composition containing compound probiotics and its application, belonging to the technical field of nutritional and health care compositions. The stomach-nourishing and protecting composition provided by the present invention comprises fructooligosaccharide, inulin, lactitol, fermented Hericium erinaceus powder, orange juice powder, konjac powder, stachyose, tremella polysaccharide, seaweed powder, fucoidan and compound probiotic powder; the compound probiotic powder is obtained by combining the bacterial powders of Lactobacillus acidophilus NCFM, Bifidobacterium animalis subsp. lactis 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. The composition provided by the present invention has a remarkable stomach-nourishing and protecting effect, can effectively improve gastric mucosal injury, and has a good market prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of nutritional and health care compositions, and particularly relates to a stomach-nourishing and stomach-protecting composition containing compound probiotics and its application. Background Art

[0002] The stomach is an organ of the human body for digestion, located under the diaphragm, connected to the esophagus above and the small intestine below. The stomach is the main organ for the digestion and absorption function of the human body. All kinds of foods need to be ground by the gastric wall (gastric mucosa) and act 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 proteins in food to facilitate the degradation by pepsin. Gastric acid can also activate pepsinogen into pepsin, thus enabling it to have the activity of decomposing proteins. Traditional Chinese medicine believes that the spleen and stomach are the foundation of acquired constitution. To keep the stomach healthy, 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 stomach-protecting composition, which contains 11 kinds of probiotic powders, has a significant effect on nourishing and protecting the stomach, and can effectively improve gastric mucosal damage.

[0004] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0005] The present invention provides a stomach-nourishing and stomach-protecting composition, which comprises the following components in parts by mass:

[0006] 35 - 50 parts of fructooligosaccharide, 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 powder, 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;

[0007] The compound bacterial powder is obtained by mixing Lactobacillus acidophilus NCFM bacterial powder, Bifidobacterium animalis subsp. lactis 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.

[0008] The Lactobacillus acidophilus NCFM powder, Bifidobacterium animalis 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 viable count ratio of (1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3):(1~3).

[0009] Preferably, the preparation method of the powder includes the following steps: inoculating the strain into a medium containing prickly pear powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder.

[0010] Preferably, the medium containing prickly pear powder includes: peptone 8~12 g / L, beef extract powder 5~15 g / L, yeast extract powder 3~7 g / L, glucose 15~25 g / L, sodium acetate 3~6 g / L, dipotassium hydrogen phosphate 1.5~2.5 g / L, ammonium citrate 1.5~2.5 g / L, magnesium sulfate 0.05~0.15 g / L, manganese sulfate 0.01~0.09 g / L, Tween 80 0.5~1.5 g / L, L-cysteine hydrochloride 0.2~0.8 g / L and prickly pear powder 8~12 g / L.

[0011] Preferably, the freeze-drying protectant includes: skim milk powder 8~12 wt%, prickly pear powder 5~15 wt%, trehalose 6~10 wt%, vitamin E 0.3~0.7 wt% and lactose 1.5~2.5 wt%.

[0012] Preferably, the mass ratio of the thallus to the freeze-drying protectant is 1:(1~3).

[0013] Preferably, the preparation method of the fermented Hericium erinaceus powder includes the following steps: inoculating Aspergillus oryzae into a fermentation medium containing Hericium erinaceus powder for fermentation, sterilizing and spray-drying the fermentation product to obtain the fermented Hericium erinaceus powder.

[0014] Preferably, the fermentation medium containing Hericium erinaceus powder includes: potato powder 150~250 g / L, Hericium erinaceus powder 150~220 g / L and glucose 15~25 g / L.

[0015] Preferably, the temperature of the fermentation is 30~33 °C, the rotation speed is 150~200 r / min, the ventilation volume is 4.0~4.8 L / min, and the time is 38~42 h.

[0016] The present invention also provides the application of the above-mentioned stomach-nourishing and stomach-protecting composition in the preparation of stomach-nourishing and stomach-protecting products.

[0017] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0018] 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

[0019] 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.

[0020] 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 11cfu / g. The compound bacterial powder of the present invention is preferably obtained by mixing each bacterial powder according to the viable count ratio of (1-3):(1-3):(1-3):(1-3):(1-3):(1-3):(1-3):(1-3):(1-3):(1-3):(1-3), more preferably obtained by mixing each bacterial powder according to an equal ratio of viable counts. The bacterial powder is preferably the freeze-dried powder of the strain.

[0021] The preservation number of Bifidobacterium lactis BR001 of the present invention is CGMCC No.23665; the preservation number of Bifidobacterium bifidum BL002 is CGMCC No.23664; the preservation number of Bifidobacterium longum subsp. longum BC012 is CGMCC No.23663; the preservation number of Lactobacillus reuteri LL029 is CGMCC No.23648; the preservation number of Lactobacillus paracasei GF045 is CGMCC No.26482; the preservation number of Lactobacillus fermentum LF028 is CGMCC No.22833; the preservation number of Lactobacillus plantarum LZ010 is CGMCC No.24258; the preservation number of Lactobacillus plantarum LZ026 is CGMCC No.22832. The Lactobacillus acidophilus NCFM is purchased from Danisco, Denmark; the Bifidobacterium animalis subsp. lactis Bb-12 is purchased from Chr. Hansen, Denmark; the Lactobacillus rhamnosus GG is purchased from Chr. Hansen, Denmark.

[0022] In the present invention, the preparation method of the bacterial powder includes the following steps: inoculating the strain into a medium containing cactus powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder.

[0023] Preferably, the preparation method of the Lactobacillus acidophilus NCFM bacterial powder includes the following steps: inoculating Lactobacillus acidophilus NCFM into a medium containing cactus powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder;

[0024] Preferably, the preparation method of the Bifidobacterium animalis subsp. lactis Bb-12 bacterial powder includes the following steps: inoculating Bifidobacterium animalis subsp. lactis Bb-12 into a medium containing cactus powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder;

[0025] Preferably, the preparation method of the Bifidobacterium lactis BR001 bacterial powder includes the following steps: inoculating Bifidobacterium lactis BR001 into a medium containing cactus powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder;

[0026] Preferably, the preparation method of the Bifidobacterium bifidum BL002 bacterial powder includes the following steps: inoculating Bifidobacterium bifidum BL002 into a medium containing cactus powder for fermentation, mixing the obtained thallus with a freeze-drying protectant, and preparing a freeze-dried powder;

[0027] Preferably, the preparation method of Bifidobacterium longum BC012 powder comprises the following steps: inoculating Bifidobacterium longum BC012 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0028] Preferably, the preparation method of Lactobacillus reuteri LL029 powder comprises the following steps: inoculating Lactobacillus reuteri LL029 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0029] Preferably, the preparation method of Lactobacillus paracasei GF045 powder comprises the following steps: inoculating Lactobacillus paracasei GF045 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0030] Preferably, the preparation method of Lactobacillus rhamnosus GG powder comprises the following steps: inoculating Lactobacillus rhamnosus GG into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0031] Preferably, the preparation method of Lactobacillus fermentum LF028 powder comprises the following steps: inoculating Lactobacillus fermentum LF028 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0032] Preferably, the preparation method of Lactobacillus plantarum LZ010 powder comprises the following steps: inoculating Lactobacillus plantarum LZ010 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder;

[0033] Preferably, the preparation method of Lactobacillus plantarum LZ026 powder comprises the following steps: inoculating Lactobacillus plantarum LZ026 into a culture medium containing prickly pear powder for fermentation, mixing the obtained bacterial cells with a lyophilization protectant, and preparing a lyophilized powder.

[0034] In the present invention, 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 in an MRS culture medium respectively to make each strain in an optimal vitality state, and a seed solution is prepared; Lactobacillus bifidus BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, and Bifidobacterium animalis subsp. lactis Bb-12 are preferably activated and expanded in a modified MRS culture medium respectively to make each strain in an optimal vitality state, and a seed solution is prepared.

[0035] The seed liquid obtained by the present invention is subjected to fermentation culture. The culture medium containing cactus powder includes: peptone 8-12 g / L, beef extract powder 5-15 g / L, yeast extract powder 3-7 g / L, glucose 15-25 g / L, sodium acetate 3-6 g / L, dipotassium hydrogen phosphate 1.5-2.5 g / L, ammonium citrate 1.5-2.5 g / L, magnesium sulfate 0.05-0.15 g / L, manganese sulfate 0.01-0.09 g / L, Tween 80 0.5-1.5 g / L, L-cysteine hydrochloride 0.2-0.8 g / L, and cactus powder 8-12 g / L. Preferably, it includes 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, dipotassium hydrogen 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, and cactus powder 10 g / L. The preparation method of the culture medium containing cactus powder includes: mixing each component of the culture medium, adding purified water, heating and boiling until completely dissolved, adjusting the pH to 6.9-7.1, and performing high-temperature sterilization. The high-temperature sterilization is preferably at 121 °C for 30 min.

[0036] The conditions for the expansion culture and fermentation culture of the present invention are static culture at 36-38 °C for 16-20 h, preferably static culture at 37 °C for 18 h. After the fermentation of the present invention is completed, the cells of each strain are centrifuged and enriched. The centrifugation conditions are preferably centrifugation at 8000 rmp for 20 min at 4 °C.

[0037] The lyoprotectant of the present invention includes skim milk powder 8-12 wt%, cactus powder 5-15 wt%, trehalose 6-10 wt%, vitamin E 0.3-0.7 wt%, and lactose 1.5-2.5 wt%. The lyoprotectant is made up to 100% with water. As an alternative embodiment, the preparation method of the lyoprotectant of the present invention includes: dissolving skim milk powder, cactus powder, trehalose, vitamin E, and lactose in water according to the mass percentage, and obtaining the lyoprotectant after high-temperature sterilization. Preferably, the mass ratio of the cells to the lyoprotectant of the present invention is 1:(1-3), preferably 1:2. The present invention mixes the cells and the lyoprotectant and then performs lyophilization. The lyophilization conditions are preferably a cold trap temperature of -75 °C and freeze-drying for 72 h. The viable count range of each bacterial powder obtained by the present invention is 1×10 11 cfu / g~9×10 11 cfu / g.

[0038] The preparation method of the fermented Hericium erinaceus powder of the present invention comprises the following steps: Aspergillus oryzae is inoculated into a fermentation medium containing Hericium erinaceus powder, followed by fermentation. The fermentation product is sterilized and spray-dried to obtain the fermented Hericium erinaceus powder. The Aspergillus oryzae of the present invention is preferably activated and cultured using a PDA medium and expanded and cultured using a PDB medium to prepare a seed solution; the activation culture conditions are preferably shaking culture at 30-34°C for 35-37 h.

[0039] The fermentation medium containing Hericium erinaceus powder of the present invention comprises: 150-250 g / L of potato powder, 150-220 g / L of Hericium erinaceus powder, and 15-25 g / L of glucose, preferably 200 g / L of potato powder, 200 g / L of Hericium erinaceus powder, and 20 g / L of glucose. The preparation method of the medium comprises: mixing each component of the medium, adding purified water, heating and boiling until completely dissolved, adjusting the pH to 5.4-5.6, and performing high-temperature sterilization. The high-temperature sterilization is preferably at 121°C for 30 min. 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 h, preferably 40 h.

[0040] The present invention sterilizes the fermentation product to remove the Aspergillus oryzae mycelium, preferably uses a filtration device to remove the Aspergillus oryzae mycelium, and spray-dries the fermentation broth. The conditions for spray drying are preferably: an inlet temperature of 190°C and an outlet temperature of 85°C to obtain the spray-dried fermented Hericium erinaceus powder.

[0041] The present invention also provides the application of the above-mentioned stomach-nourishing and stomach-protecting composition in the preparation of stomach-nourishing and stomach-protecting products, and the products are preferably drugs or health products.

[0042] Next, the technical solutions in the present invention will be clearly and completely described in conjunction with the embodiments in the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present invention without making creative efforts shall fall within the protection scope of the present invention.

[0043] In the specific embodiments of the present invention:

[0044] The Hericium erinaceus powder and cactus powder are purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0045] Preparation of the medium:

[0046] (1) PDA medium: potato powder 200 g / L, glucose 20 g / L, agar 20 g / L; Mix each component of the medium, add purified water, heat to boiling until completely dissolved, sterilize at 121 °C for 30 min to obtain the PDA medium.

[0047] (2) PDB medium (basic fermentation medium): potato powder 200 g / L, glucose 20 g / L; Mix each component of the medium, add purified water, heat to boiling until completely dissolved, adjust the pH to 5.5, sterilize at 121 °C for 30 min to obtain the PDB medium.

[0048] (3) PDC medium (Hericium erinaceus fermentation medium): potato powder 200 g / L, Hericium erinaceus powder 200 g / L, glucose 20 g / L; Mix each component of the medium, add purified water, heat to boiling until completely dissolved, adjust the pH to 5.5, sterilize at 121 °C for 30 min to obtain the PDC medium.

[0049] (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, dipotassium hydrogen 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 each component of the medium, add purified water, heat to boiling until completely dissolved, adjust the pH to 7.0, sterilize at 121 °C for 30 min to obtain the XRZ medium.

[0050] (5) MRS medium: The MRS medium is purchased from Beijing Land Bridge Technology Co., Ltd. Weigh 55.2 g of MRS medium raw materials into 1 L of distilled water, heat to boiling until completely dissolved, sterilize at 121 °C under high pressure for 15 min to obtain the MRS medium.

[0051] (6) Modified MRS medium: The MRS medium is purchased from Beijing Land Bridge Technology Co., Ltd. Weigh 55.2 g of MRS medium raw materials and 0.5 g of L-cysteine hydrochloride into 1 L of distilled water, heat to boiling until completely dissolved, sterilize at 121 °C under high pressure for 15 min to obtain the modified MRS medium.

[0052] (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, dipotassium hydrogen 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; each component of the medium was mixed, purified water was added, heated and boiled until completely dissolved, the pH was adjusted to 7.0, sterilized at 121 °C for 30 min, and the HTG medium was prepared.

[0053] (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, dipotassium hydrogen 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; each component of the medium was mixed, purified water was added, heated and boiled until completely dissolved, the pH was adjusted to 7.0, sterilized at 121 °C for 30 min, and the HTG-Q control medium was prepared.

[0054] The strains used in the specific embodiments of the present invention are as follows: Aspergillus oryzae was purchased from Shandong Jining Xiangyuan Biotechnology Co., Ltd.; Lactobacillus acidophilus LS001, with the preservation number of CGMCC No. 23649; Bifidobacterium lactis BR001, with the preservation number of CGMCC No. 23665; Bifidobacterium bifidum BL002, with the preservation number of CGMCC No. 23664; Bifidobacterium longum BC012, with the preservation number of CGMCC No. 23663; Lactobacillus reuteri LL029, with the preservation number of CGMCC No. 23648; Lactobacillus paracasei GF045, with the preservation number of CGMCC No. 26482; Lactobacillus paracasei GF027, with the preservation number of CGMCC No. 22831; Lactobacillus fermentum LF028, with the preservation number of CGMCC No. 22833; Lactobacillus plantarum LZ010, with the preservation number of CGMCC No. 24258; Lactobacillus plantarum LZ026, with the 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.

[0055] In the following examples, unless otherwise specified, all are conventional methods.

[0056] The materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial channels.

[0057] Example 1

[0058] 1. Explore the polysaccharide and β-glucan contents in the fermented liquid of Hericium erinaceus powder obtained by extraction at different fermentation times:

[0059] (1) Tube slant activation culture: Aspergillus oryzae was inoculated on a PDA slant medium and cultured at a constant temperature of 30 °C for 6 days.

[0060] (2) Preparation of spore suspension: Add 5 mL of sterile water to the slant of the activated strain, gently scrape and wash with an inoculation loop, pour it into a sterilized Erlenmeyer flask containing glass beads, and use the glass beads to fully disperse the spores to prepare a spore suspension of 1×10 7 spores / mL for standby.

[0061] (3) Seed shake flask culture: Inoculate 4 mL of spore suspension into a 250 mL Erlenmeyer flask containing 50 mL of PDB medium, and place it in a constant temperature shaker at 32 °C and 180 r / min for 36 h to obtain a seed solution.

[0062] (4) Liquid shake flask culture: Add the Aspergillus oryzae seed solution to 10 250 mL Erlenmeyer flasks containing 50 mL of PDC medium at an inoculation amount of 1% (v / v), and place them in a constant temperature shaker at 32 °C and 180 r / min. Take one bottle for detection at 0 h, 8 h, 16 h, 24 h, 32 h, 40 h, 48 h, 56 h, 64 h, and 72 h respectively. This experiment was repeated three times.

[0063] (5) Detection of β-glucan content: Determined using the Megazyme kit (K-YBGL). Calculate the β-glucan yield according to formula (1):

[0064] Formula (1).

[0065] (6) Polysaccharide extraction: Take a quantitative fermented liquid and centrifuge it (4000 g, 30 min), retain the supernatant, add absolute ethanol (the volume ratio of the supernatant to absolute ethanol is 25:75) for alcohol precipitation, let it stand for 24 h and then centrifuge it (4200 g, 30 min), retain the precipitate; dissolve the above precipitate in water at 65 °C and then freeze-dry it to obtain Hericium erinaceus extract. Use the phenol-sulfuric acid method to determine the polysaccharide content in the extracted Hericium erinaceus extract, and calculate the polysaccharide yield through formula (2):

[0066] Formula (2).

[0067] The polysaccharide and β-glucan contents obtained by extraction at different fermentation times are shown in Table 1. The "*" in the table indicates: compared with 0 h, P <0.05.

[0068] Table 1 Polysaccharide and β-glucan contents in each group

[0069]

[0070] The results showed that during the fermentation of Aspergillus oryzae, with the prolongation of fermentation time, the contents of polysaccharides and β-glucans in Hericium erinaceus gradually increased and reached the highest at 40 h, and then decreased from 48 h to 72 h. The possible reasons for the changes in the contents of polysaccharides and β-glucans in Hericium erinaceus are as follows: various enzymes secreted by Aspergillus oryzae, such as amylase, glucoamylase, cellulase, phytase, pectinase, etc., hydrolyzed Hericium erinaceus in the early stage of fermentation, promoting the increase in the contents of polysaccharides and β-glucans in Hericium erinaceus. In the later stage of fermentation, as Aspergillus oryzae grew, it decomposed and utilized the polysaccharides and β-glucans in Hericium erinaceus for consumption.

[0071] 2. Explore the differences between fermented Hericium erinaceus powder and unfermented Hericium erinaceus powder

[0072] (1) Preparation of fermented Hericium erinaceus powder: Prepare Aspergillus oryzae seed liquid by the same method as in step 1. The Aspergillus oryzae seed liquid was inoculated into a 5 L fermenter with PDC medium at an inoculation amount of 1% (v / v), and fermented at 32 °C, 180 r / min, and an aeration rate of 4.5 L / min for 40 h and then terminated. After removing the mycelium of Aspergillus oryzae using a 60-mesh filtration device, the fermentation broth was spray-dried at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain the spray-dried Hericium erinaceus powder.

[0073] (2) Preparation of unfermented Hericium erinaceus powder: The sterilized PDC medium was directly spray-dried at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain the spray-dried Hericium erinaceus powder without fermentation by Aspergillus oryzae.

[0074] Using the same method as in step 1, the contents of β-glucan and polysaccharide in fermented Hericium erinaceus powder and unfermented Hericium erinaceus powder were measured respectively. The test results are shown in Table 2, where "**" in the table indicates: compared with unfermented Hericium erinaceus powder, P <0.01.

[0075] Table 2 Contents of polysaccharides and β-glucans in fermented Hericium erinaceus powder and unfermented Hericium erinaceus powder

[0076]

[0077] The results showed that after fermentation by Aspergillus oryzae, the contents of polysaccharides and β-glucans in Hericium erinaceus powder were greatly increased.

[0078] Example 2

[0079] 1. Effect of adding cactus powder on lactic acid bacteria

[0080] Cactus powder was used to culture lactic acid bacteria to detect whether cactus powder is beneficial to the growth of lactic acid bacteria.

[0081] MRS medium was used to culture Lactobacillus: 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 respectively inoculated into sterilized MRS medium, statically cultured at 37 °C for 18 h, continuously passaged and activated for 2 times, and then inoculated into sterilized MRS medium at 3% (v / v) again, and statically cultured at 37 °C for 18 h as the seed liquid for standby.

[0082] Modified MRS medium was used to culture Bifidobacterium: Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum BC012, Bifidobacterium animalis subsp. lactis Bb-12; the above strains were respectively inoculated into sterilized modified MRS medium, anaerobically and statically cultured at 37 °C for 18 h, continuously passaged and activated for 2 times, and then inoculated into sterilized MRS medium at 3% (v / v) again, and statically cultured at 37 °C for 18 h as the seed liquid for standby.

[0083] After the 13 strains were expanded in culture, the seed liquid was inoculated into XRZ medium at an inoculation amount of 3% (v / v) respectively, and after anaerobic static culture at 37 °C for 16 h, its pH, OD value and viable count were detected. The results are shown in Table 3.

[0084] Table 3 pH, OD value and viable count of 13 strains

[0085]

[0086] 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, with a higher pH, a lower OD value and a smaller viable count. Cactus powder had an inhibitory effect on the growth of these two lactic acid bacteria; 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, Lactobacillus rhamnosus GG, a total of 11 strains could grow normally in XRZ medium, and the viable count reached 10 9 Above. Thereafter, the experiment was carried out with 11 strains, removing Lactobacillus acidophilus LS001 and Lactobacillus paracasei GF027.

[0087] 2. Experiment on the effect of Hericium erinaceus powder on lactic acid bacteria

[0088] 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.

[0089] The seed solutions 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 subsp. lactis Bb-12 were prepared by the same method as in Step 1.

[0090] The seed solutions of the 11 strains were inoculated into HTG and HTG-Q media at an inoculation amount of 3% (v / v), and after anaerobic static culture at 37 °C for 16 h, the viable cell count was detected. The results are shown in Table 4.

[0091] Table 4 pH, OD value, and viable cell count of 11 strains

[0092]

[0093] By comparing the two media with and without Hericium erinaceus powder added, the results showed that Hericium erinaceus powder is beneficial to the increase in the viable cell count of lactic acid bacteria. The reason may be that Hericium erinaceus contains various bioactive substances such as polysaccharides, oligosaccharides, sterols, terpenoids, phenols, adenosine, unsaturated fatty acids, proteins, and polypeptides, which have a good growth-promoting effect on the bacteria. Therefore, adding Hericium erinaceus powder is beneficial to the growth of lactic acid bacteria.

[0094] 3. Comparison between cactus powder lactic acid bacteria powder and ordinary lactic acid bacteria powder

[0095] (1) Preparation of cactus powder lactic acid bacteria powder:

[0096] The seed solutions 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 were prepared in the same way as in Step 1. The seed solutions were inoculated into XRZ medium at an inoculation amount of 3% (v / v), and after anaerobic static culture at 37 °C for 16 h, the cells of each strain were enriched by centrifugation at 8000 rmp for 20 min at 4 °C.

[0097] The cells of each strain were thoroughly mixed with the cryoprotectant at a mass ratio of 1:2, and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder of each strain, and the viable cell count of each freeze-dried powder was performed.

[0098] The cryoprotectant consists of: skim milk powder 10 wt%, cactus powder 10 wt%, trehalose 8 wt%, vitamin E 0.5 wt%, lactose 2 wt%, and the rest is water. The skim milk powder, cactus powder, trehalose, vitamin E and lactose are dissolved in water according to the mass percentage, and after sterilization at 110 °C for 20 min, the freeze-drying cryoprotectant is obtained.

[0099] (2)Preparation of common lactic acid bacteria powder

[0100] The seed solutions 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. Bb-12, and Lactobacillus rhamnosus GG are prepared in the same way as in step 1. The seed solutions are respectively inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h, and then the cells of each strain are enriched by centrifugation at 8000 rmp for 20 min at 4 °C.

[0101] The cells of each strain are thoroughly mixed with the cryoprotectant at a mass ratio of 1:2, and freeze-dried at -75 °C for 72 h to obtain the freeze-dried powder of each strain, and the viable cell count of each freeze-dried powder is carried out.

[0102] The cryoprotectant consists of: skim milk powder 10 wt%, maltodextrin 10 wt%, trehalose 8 wt%, vitamin E 0.5 wt%, lactose 2 wt%, and the rest is water. The skim milk powder, maltodextrin, trehalose, vitamin E and lactose are dissolved in water according to the mass percentage, and after sterilization at 110 °C for 20 min, the freeze-drying cryoprotectant is obtained.

[0103] Compare the viable cell counts of the cactus powder lactic acid bacteria powder and the common lactic acid bacteria powder, and the results are shown in Table 5.

[0104] Table 5 Viable cell counts of different bacterial powders prepared from 11 strains

[0105]

[0106] The results show that the viable cell counts of the freeze-dried powders of the 11 strains are all above 10 11 and there is no difference, and the cactus powder has no adverse effect on the strains.

[0107] Example 3

[0108] A stomach-nourishing and stomach-protecting composition is obtained by mixing 50 g of fructooligosaccharide, 20 g of inulin, 15 g of lactitol, 5 g of fermented Hericium erinaceus powder, 2 g of orange juice powder, 2 g of konjac powder, 0.5 g of stachyose, 0.5 g of tremella polysaccharide, 0.5 g of seaweed powder, 2.5 g of fucoidan, and 2 g of compound bacterial powder.

[0109] Preparation of fermented Hericium erinaceus powder:

[0110] (1) Tube slant activation culture: Aspergillus oryzae was inoculated on a PDA slant medium and cultured at a constant temperature of 30 °C for 6 days.

[0111] (2) Preparation of spore suspension: 5 mL of sterile water was added to the slant of the activated strain, gently scraped with an inoculation loop, and poured into a sterilized Erlenmeyer flask containing glass beads. The spores were fully dispersed with the glass beads to prepare a spore suspension of 1×10 7 spores / mL for standby.

[0112] (3) Seed shake flask culture: 4 mL of spore suspension was inoculated into a 250 mL Erlenmeyer flask containing 50 mL of PDB medium and cultured in a constant temperature shaker at 32 °C and 180 r / min for 36 h.

[0113] (4) The Aspergillus oryzae seed liquid was inoculated into a 5 L PDC medium fermentation tank at an inoculation amount of 1%. Fermentation was carried out at 32 °C, 180 r / min, and an aeration rate of 4.5 L / min. After 40 h of fermentation, it was terminated. After removing the Aspergillus oryzae mycelium with a 60-mesh filtration device, the fermentation broth was spray-dried at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain the spray-dried Hericium erinaceus powder.

[0114] Preparation of compound bacterial powder:

[0115] Seed liquids of Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum subsp. 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 were respectively prepared according to the steps of Step 1 of Example 2.

[0116] (1) Preparation of freeze-dried powder of Bifidobacterium lactis BR001: The seed liquid of Bifidobacterium lactis BR001 was inoculated into the XRZ medium at an inoculation amount of 3% (v / v) and anaerobically statically cultured at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protectant at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0117] (2)Preparation of Bifidobacterium bifidum BL002 freeze-dried powder: The seed liquid of Bifidobacterium bifidum BL002 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0118] (3)Preparation of Bifidobacterium longum subsp. longum BC012 freeze-dried powder: The seed liquid of Bifidobacterium longum subsp. longum BC012 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0119] (4)Preparation of Lactobacillus reuteri LL029 freeze-dried powder: The seed liquid of Lactobacillus reuteri LL029 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0120] (5)Preparation of Lactobacillus paracasei GF045 freeze-dried powder: The seed liquid of Lactobacillus paracasei GF045 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0121] (6)Preparation of Lactobacillus fermentum LF028 freeze-dried powder: The seed liquid of Lactobacillus fermentum LF028 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0122] (7)Preparation of Lactobacillus plantarum LZ010 freeze-dried powder: The seed liquid of Lactobacillus plantarum LZ010 was inoculated into XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. 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 freeze-dried powder.

[0123] Preparation of freeze-dried powder of Lactobacillus plantarum LZ026: The seed liquid of Lactobacillus plantarum LZ026 was inoculated into the XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells and the cryoprotectant 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 freeze-dried powder.

[0124] Preparation of freeze-dried powder of Lactobacillus acidophilus NCFM: The seed liquid of Lactobacillus acidophilus NCFM was inoculated into the XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells and the cryoprotectant 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 freeze-dried powder.

[0125] Preparation of freeze-dried powder of Bifidobacterium animalis subsp. lactis Bb-12: The seed liquid of Bifidobacterium animalis subsp. lactis Bb-12 was inoculated into the XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells and the cryoprotectant 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 freeze-dried powder.

[0126] Preparation of freeze-dried powder of Lactobacillus rhamnosus GG: The seed liquid of Lactobacillus rhamnosus GG was inoculated into the XRZ medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells and the cryoprotectant 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 freeze-dried powder.

[0127] The cryoprotectant is: skim milk powder 10 wt%, cactus powder 10 wt%, trehalose 8 wt%, vitamin E 0.5 wt%, lactose 2 wt%, and the rest is water. The skim milk powder, cactus powder, trehalose, vitamin E and lactose were dissolved in water according to the mass percentage, and sterilized at 110°C for 20 min to obtain the freeze-drying protectant.

[0128] (12)The composite bacterial powder is prepared by fully mixing freeze-dried Lactobacillus acidophilus NCFM, freeze-dried Bifidobacterium animalis subsp. lactis Bb-12, freeze-dried Bifidobacterium lactis BR001, freeze-dried Bifidobacterium bifidum BL002, freeze-dried Bifidobacterium longum BC012, freeze-dried Lactobacillus reuteri LL029, freeze-dried Lactobacillus paracasei GF045, freeze-dried Lactobacillus rhamnosus GG, freeze-dried Lactobacillus fermentum LF028, freeze-dried Lactobacillus plantarum LZ010, and freeze-dried Lactobacillus plantarum LZ026 according to equal viable counts (the order of magnitude of the viable count is 10 11 cfu / g).

[0129] Example 4

[0130] A stomach-nourishing and stomach-protecting composition is obtained by mixing 35 g of fructooligosaccharide, 25 g of inulin, 15 g of lactitol, 5 g of fermented Hericium erinaceus powder, 6 g of orange juice powder, 2 g of konjac powder, 1.5 g of stachyose, 1.5 g of tremella polysaccharide, 1.5 g of seaweed powder, 3.5 g of fucoidan, and 4 g of composite bacterial powder.

[0131] The preparation method of the fermented Hericium erinaceus powder and the preparation method of the composite bacterial powder are the same as those in Example 3.

[0132] Example 5

[0133] A stomach-nourishing and stomach-protecting composition is obtained by mixing 35 g of fructooligosaccharide, 20 g of inulin, 25 g of lactitol, 5 g of fermented Hericium erinaceus powder, 3 g of orange juice powder, 4 g of konjac powder, 1.0 g of stachyose, 0.8 g of tremella polysaccharide, 1.2 g of seaweed powder, 2 g of fucoidan, and 3 g of composite bacterial powder.

[0134] The preparation method of the fermented Hericium erinaceus powder and the preparation method of the composite bacterial powder are the same as those in Example 3.

[0135] Example 6

[0136] A stomach-nourishing and stomach-protecting composition is obtained by mixing 35.8 g of fructooligosaccharide, 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 powder, 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.

[0137] The preparation method of the fermented Hericium erinaceus powder and the preparation method of the composite bacterial powder are the same as those in Example 3.

[0138] Example 7

[0139] A stomach-nourishing and stomach-protecting composition is obtained by mixing 40 g of fructooligosaccharide, 22 g of inulin, 19.5 g of lactitol, 7 g of fermented Hericium erinaceus powder, 2 g of orange juice powder, 3 g of konjac powder, 0.5 g of stachyose, 0.5 g of tremella polysaccharide, 0.6 g of seaweed powder, 2.4 g of fucoidan, and 2.5 g of compound bacterial powder.

[0140] The preparation methods of the fermented Hericium erinaceus powder and the compound bacterial powder are the same as those in Example 3.

[0141] Example 8

[0142] A stomach-nourishing and stomach-protecting composition is obtained by mixing 42 g of fructooligosaccharide, 20 g of inulin, 21 g of lactitol, 6 g of fermented Hericium erinaceus powder, 2 g of orange juice powder, 2 g of konjac powder, 1.2 g of stachyose, 1.1 g of tremella polysaccharide, 0.7 g of seaweed powder, 2 g of fucoidan, and 2 g of compound bacterial powder.

[0143] The preparation methods of the fermented Hericium erinaceus powder and the compound bacterial powder are the same as those in Example 3.

[0144] Comparative Example 1

[0145] A stomach-nourishing and stomach-protecting composition is obtained by mixing 50 g of fructooligosaccharide, 20 g of inulin, 15 g of lactitol, 5 g of Hericium erinaceus powder, 2 g of orange juice powder, 2 g of konjac powder, 0.5 g of stachyose, 0.5 g of tremella polysaccharide, 0.5 g of seaweed powder, 2.5 g of fucoidan, and 2 g of cactus powder.

[0146] Preparation method of Hericium erinaceus powder: Directly spray-dry the sterilized PDC medium at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain spray-dried Hericium erinaceus powder without Aspergillus oryzae fermentation.

[0147] Comparative Example 2

[0148] A stomach-nourishing and stomach-protecting composition is obtained by mixing 35.8 g of fructooligosaccharide, 21 g of inulin, 17 g of lactitol, 10 g of Hericium erinaceus powder, 4 g of orange juice powder, 3 g of konjac powder, 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 compound bacterial powder.

[0149] Preparation method of Hericium erinaceus powder: Directly spray-dry the sterilized PDC medium at an inlet temperature of 190 °C and an outlet temperature of 85 °C to obtain spray-dried Hericium erinaceus powder without Aspergillus oryzae fermentation.

[0150] Preparation of compound bacterial powder:

[0151] Bifidobacterium lactis BR001, Bifidobacterium bifidum BL002, Bifidobacterium longum subsp. 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 were respectively prepared into seed solutions according to the steps of Step 1 of Example 2.

[0152] (1)Preparation of freeze-dried powder of Bifidobacterium lactis BR001: The seed solution of Bifidobacterium lactis BR001 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. Then, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the freeze-dried powder.

[0153] (2)Preparation of freeze-dried powder of Bifidobacterium bifidum BL002: The seed solution of Bifidobacterium bifidum BL002 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. Then, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the freeze-dried powder.

[0154] (3)Preparation of freeze-dried powder of Bifidobacterium longum subsp. longum BC012: The seed solution of Bifidobacterium longum subsp. longum BC012 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. Then, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the freeze-dried powder.

[0155] (4)Preparation of freeze-dried powder of Lactobacillus reuteri LL029: The seed solution of Lactobacillus reuteri LL029 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. Then, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the freeze-dried powder.

[0156] (5)Preparation of freeze-dried powder of Lactobacillus paracasei GF045: The seed solution of Lactobacillus paracasei GF045 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37°C for 16 h. Then, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4°C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75°C for 72 h to obtain the freeze-dried powder.

[0157] (6) Preparation of freeze-dried powder of Lactobacillus fermentum LF028: The seed solution of Lactobacillus fermentum LF028 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0158] (7) Preparation of freeze-dried powder of Lactobacillus plantarum LZ010: The seed solution of Lactobacillus plantarum LZ010 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0159] (8) Preparation of freeze-dried powder of Lactobacillus plantarum LZ026: The seed solution of Lactobacillus plantarum LZ026 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0160] (9) Preparation of freeze-dried powder of Lactobacillus acidophilus NCFM: The seed solution of Lactobacillus acidophilus NCFM was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0161] (10) Preparation of freeze-dried powder of Bifidobacterium animalis subsp. lactis Bb-12: The seed solution of Bifidobacterium animalis subsp. lactis Bb-12 was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0162] (11) Preparation of freeze-dried powder of Lactobacillus rhamnosus GG: The seed solution of Lactobacillus rhamnosus GG was inoculated into MRS medium at an inoculation amount of 3% (v / v) and cultured anaerobically and statically at 37 °C for 16 h. After that, the cells were enriched by centrifugation at 8000 rmp for 20 min at 4 °C. The cells were thoroughly mixed with the protective agent at a mass ratio of 1:2 and freeze-dried at a cold trap temperature of -75 °C for 72 h to obtain the freeze-dried powder.

[0163] The cryoprotectant is: skim milk powder 10 wt%, maltodextrin 10 wt%, trehalose 8 wt%, vitamin E 0.5 wt%, lactose 2 wt%, and the rest is water. The skim milk powder, maltodextrin, trehalose, vitamin E and lactose are dissolved in water according to the mass percentage, and after sterilization at 110 °C for 20 min, a lyophilization cryoprotectant is obtained.

[0164] (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 are fully mixed according to equal viable counts to obtain a compound bacterial powder (the order of magnitude of the viable count is 10 11 cfu / g).

[0165] Comparative Example 3

[0166] A stomach-nourishing and stomach-protecting composition is obtained by mixing 35 g of fructooligosaccharide, 20 g of inulin, 25 g of lactitol, 5 g of Hericium erinaceus powder, 3 g of orange juice powder, 4 g of konjac powder, 1.0 g of stachyose, 0.8 g of tremella polysaccharide, 1.2 g of seaweed powder, 2 g of fucoidan, and 3 g of compound bacterial powder.

[0167] Preparation method of Hericium erinaceus powder: directly perform spray drying on the sterilized PDC medium, with an inlet temperature of 190 °C and an outlet temperature of 85 °C, to obtain spray-dried Hericium erinaceus powder without Aspergillus oryzae fermentation.

[0168] The preparation method of the compound bacterial powder is the same as that in Example 3.

[0169] Comparative Example 4

[0170] A stomach-nourishing and stomach-protecting composition is obtained by mixing 40 g of fructooligosaccharide, 22 g of inulin, 19.5 g of lactitol, 7 g of fermented Hericium erinaceus powder, 2 g of orange juice powder, 3 g of konjac powder, 0.5 g of stachyose, 0.5 g of tremella polysaccharide, 0.6 g of seaweed powder, 2.4 g of fucoidan, and 2.5 g of compound bacterial powder.

[0171] The preparation method of the fermented Hericium erinaceus powder is the same as that in Example 3.

[0172] The preparation method of the compound bacterial powder is the same as that in Comparative Example 2.

[0173] Comparative Example 5

[0174] A stomach-nourishing and stomach-protecting composition is obtained by mixing 42 g of fructooligosaccharide, 20 g of inulin, 21 g of lactitol, 6 g of Hericium erinaceus powder, 2 g of orange juice powder, 2 g of konjac powder, 1.2 g of stachyose, 1.1 g of tremella polysaccharide, 0.7 g of seaweed powder, 2 g of fucoidan, and 2 g of compound bacterial powder.

[0175] The preparation method of the fermented Hericium erinaceus powder is the same as that in Example 3.

[0176] Compound bacterial powder: It is obtained by mixing Lactobacillus acidophilus NCFM bacterial powder, Bifidobacterium animalis subsp. lactis Bb-12 bacterial powder, Bifidobacterium bifidum BL002 bacterial powder, Bifidobacterium longum BC012 bacterial powder, Lactobacillus paracasei GF045 bacterial powder, Lactobacillus rhamnosus GG bacterial powder, Lactobacillus fermentum LF028 bacterial powder, and Lactobacillus plantarum LZ010 bacterial powder (the order of magnitude of the viable bacteria count is 10 11 cfu / g). The compound bacterial powder in this comparative example does not contain Bifidobacterium lactis BR001 bacterial powder, Lactobacillus reuteri LL029 bacterial powder, and Lactobacillus plantarum LZ026 bacterial powder. The preparation method of each bacterial powder is the same as that in Example 3.

[0177] Test Example 1

[0178] Animal acute gastric mucosal injury experiment: The eighteenth item "Test method for assisting in protecting gastric mucosa" in the "Functional Test and Evaluation Method of Health Foods (2022 Edition)" is adopted.

[0179] Experimental animals: Wistar healthy rats, male, with a body weight of 180 - 220 g, a total of 140 rats.

[0180] The experiment is set up as experimental groups 1 - 6, control groups 7 - 11, blank control group, model control group, and positive control group, a total of 14 groups, with 10 animals in each group. They are gavaged once a day, with a gavage volume of 1.0 mL / 100 g, for 30 consecutive days. Experimental groups 1 - 6 correspond to the compositions prepared in Example 3, Example 4, Example 5, Example 6, Example 7, and Example 8 respectively. Control groups 7 - 11 represent the compositions prepared in Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, and Comparative Example 5 respectively. The gavage dose of each group is 50 mg / kg each time (dissolved in physiological saline for gavage); the positive control group is gavaged with cimetidine, and the gavage dose of cimetidine is 0.04 g / kg each time (dissolved in physiological saline for gavage); the blank control group and the model control group are given an equal volume of physiological saline.

[0181] After 30 days of intragastric administration to each group, all animals were strictly fasted for 24 h (water not prohibited), and the test substance was also prohibited from being given during this period. Except for the blank control, all experimental group animals were given 1.0 mL / animal of absolute ethanol. One hour later, the animals were sacrificed, the intact stomach was exposed, the pylorus was ligated, an appropriate amount of 10% formalin solution was perfused, fixed for 20 min, then cut along the greater curvature of the stomach, the gastric contents were washed, the gastric mucosa was unfolded, and the length and width of the bleeding points or bleeding bands were measured with a vernier caliper under a stereoscopic dissecting microscope. Since the width represents the severity of the injury much more than the length, the score was doubled. The scoring criteria are shown in Table 6, and the experimental results are shown in Table 7.

[0182] Table 6 Gross observation scoring criteria for acute absolute ethanol injury

[0183]

[0184] Table 7 Gastric injury score results of each group

[0185]

[0186] Note: Compared with the control group, * P <0.05; ** P <0.01. Compared with the model group, # P <0.05; ## P <0.01.

[0187] The results showed that compared with the control group, the alcohol model of acute gastric mucosal injury in rats was successfully established; compared with the model group, the compound ratios of experimental groups 1 - 6 could significantly reduce gastric mucosal injury, and the difference was extremely significant ( P <0.01); compared with the model group, the compound ratios of comparison groups 7 - 11 could reduce gastric mucosal injury, and the difference was 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.

[0188] Test Example 2

[0189] Crowd trial experiment:

[0190] 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.

[0191] 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.

[0192] 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.

[0193] Table 8 Symptom severity classification table for human food trial

[0194]

[0195] Table 9 Questionnaire results after 30 days of consumption

[0196]

[0197] It is known from the questionnaire answers of the subjects that the composition formula of the present invention can effectively improve the gastric discomfort of the human subjects: among them, the people with acid reflux, abdominal distension and less food intake have been significantly improved; the people with belching are the second; the people with stomachache have the lowest improvement rate. The total improvement rate of five indicators reaches 73.28%, indicating that the formula of the present invention can bring good improvement to the people with gastric mucosal injury.

[0198] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope 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 powder, Bifidobacterium animalis 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 according to the number of viable cells (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3): (1 to 3); The preparation method of bacterial powder comprises the following steps: inoculating a strain into a culture medium containing cactus powder for fermentation, mixing the fermented bacterial bodies with a freeze-drying protective agent, and preparing freeze-dried powder; 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; 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 and spray-drying the fermentation product to obtain the fermented hericium erinaceus powder; the fermentation medium containing hericium erinaceus powder comprises: 150 to 250 g / L of potato flour, 150 to 220 g / L of hericium erinaceus powder and 15 to 25 g / L of glucose; the fermentation temperature is 30 to 33° C., the rotation speed is 150 to 200 r / min, the ventilation volume is 4.0 to 4.8 L / min, and the time is 38 to 42 h.

2. The stomach nourishing and stomach protecting composition according to claim 1, characterized in that: The freeze-drying protective agent 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.

3. The stomach nourishing and stomach protecting composition according to claim 1, characterized in that: The mass ratio of the bacterial cells to the lyophilization protective agent is 1: (1 to 3).

4. Use of the stomach nourishing and stomach protecting composition according to any one of claims 1 to 3 in the preparation of stomach nourishing and stomach protecting products.

Images