A fermented black fungus beverage with laxative function and its preparation method

Abstract

The application discloses a fermented black fungus beverage with the functions of moistening intestines and defecating and a preparation method thereof, and belongs to the technical field of food processing. The fermented black fungus beverage is prepared by inoculating probiotics into black fungus extract, and further compounding the black fungus extract with natural extracts such as inulin and xylo-oligosaccharide, so that the fermented black fungus beverage has soft sour taste and enhanced functions of moistening intestines and defecating. The functional properties of the fermented black fungus beverage are characterized by animal experiments, and a formula and a production process of the fermented black fungus beverage with strong functionality and good taste are obtained. The fermented black fungus beverage prepared by the application has better functions of moistening intestines and defecating through the compounding of inulin and xylo-oligosaccharide, and has better functions of moistening intestines and defecating under the premise of controlling the dosage of additives.

Description

Technical Field

[0001] This invention relates to a fermented black fungus beverage with laxative function and its preparation method, belonging to the field of food processing technology. Background Technology

[0002] Black fungus (Auricularia auricular-judae), also known as wood ear or wood fungus, is a large edible fungus belonging to the subphylum Basidiomycota, class Agaricomycetes, family Auriculariaceae, and genus Auricularia. my country is the world's largest producer of black fungus, accounting for approximately 90% of global production annually, mainly distributed in Heilongjiang, Jilin, Shaanxi, Guangxi, Sichuan, and Guizhou provinces. Black fungus has a thin, elastic texture, a rich flavor, and is very popular among consumers. Studies have shown that black fungus promotes intestinal peristalsis, increases stool water content, and has a certain therapeutic effect on constipation. Black fungus contains various active substances, such as dietary fiber, polysaccharides, and plant colloids. The dietary fiber in black fungus has good water-holding capacity and swelling power, which increases stool wet weight, accelerates intestinal motility, and stimulates the secretion of gastrointestinal hormones, thus achieving a laxative effect. Black fungus polysaccharides are natural plant polysaccharides with bioactive functions such as regulating human immunity, anti-oxidation, anticoagulation, and promoting gastrointestinal motility. In addition, compared with drug treatment, natural active substances have a lower incidence of adverse reactions in relieving constipation.

[0003] Currently, the main black fungus products on the market include steamed buns, flour, biscuits, and beverages, while fermented black fungus beverages are rarely reported. Probiotic fermentation can effectively improve the flavor of black fungus, remove the earthy taste, increase acidity, and create a desired sweet and sour taste. It also hydrolyzes the proteins and polysaccharides in black fungus, increasing the release and solubility of polysaccharides. However, there are few reports on lactic acid bacteria fermentation processes for black fungus. How to increase the content and efficacy of soluble black fungus polysaccharides while maintaining a good taste remains a technical challenge for the industry. Summary of the Invention

[0004] This invention involves fermenting black fungus extract with a compound bacteria (Lactobacillus bulgaricus, Streptococcus thermophilus, and Bifidobacterium longum) to better hydrolyze and extract the effective component, black fungus polysaccharides. The fermentation process gives the product a mellow, pleasantly sour taste. The addition of small pieces of black fungus pulp juice enhances its crisp texture and consumer trust. Furthermore, the fermented broth rich in black fungus polysaccharides obtained by this invention is combined with natural extracts such as inulin and xylooligosaccharides to strengthen its laxative function, resulting in a palatable and highly functional fermented black fungus beverage.

[0005] This invention provides a fermented black fungus beverage with laxative function and its preparation method. The method involves preparing black fungus fermentation liquid and black fungus pulp juice separately, mixing them, and then adding inulin, xylooligosaccharides and other raw materials for blending.

[0006] This invention provides a method for preparing a fermented black fungus beverage that promotes bowel movement and relieves constipation, comprising the following steps:

[0007] (1) Preparation of black fungus fermentation liquid

[0008] After the black fungus is crushed, it is soaked in water, whey protein powder and concentrated pear juice are added, and it is boiled to sterilize. After cooling, a compound bacterial strain is added and fermented. The clear liquid is filtered to obtain the black fungus fermentation liquid. The compound bacterial strain is Lactobacillus bulgaricus, Bifidobacterium longum and Streptococcus thermophilus.

[0009] (2) Preparation of black fungus pulp juice

[0010] Crush the black fungus, soak it in water, add sugar and boil it to get black fungus pulp juice;

[0011] (3) Preparation of black fungus beverage for promoting bowel movement

[0012] The fermented black fungus liquid obtained in step (1) and the black fungus pulp juice obtained in step (2) are mixed, and xylooligosaccharides, inulin and sodium benzoate are added. After filling, sealing and sterilization, the fermented black fungus beverage is obtained.

[0013] In one embodiment of the present invention, the method for preparing the black fungus fermentation liquid is as follows:

[0014] Soak black fungus powder in water at a ratio of 1:80 to 1:150 for 30 to 60 minutes, then mash it into a paste. Add 0.5 to 2 g of whey protein powder and 1 to 7 g of concentrated pear juice per 100 mL. Sterilize by simmering at 85 to 95°C for 20 to 30 minutes. Add 1% to 3% of a compound bacterial strain (a combination of Lactobacillus bulgaricus, Bifidobacterium longum, and Streptococcus thermophilus) and incubate at 25 to 45°C. The fermentation endpoint is reached when the total acidity reaches 2.50 to 3.50, yielding the black fungus fermentation broth.

[0015] In one embodiment of the present invention, in step (1), black fungus is pulverized to 80-300 mesh, and soaked in water at a ratio of (1:80) to (1:150) for 20-60 minutes. After pulping, 0.5g / 100mL to 2g / 100mL of whey protein and 1g / 100mL to 7g / 100mL of concentrated pear juice are added.

[0016] In one embodiment of the present invention, the composite bacterial strain is obtained by mixing Lactobacillus bulgaricus, Bifidobacterium longum, and Streptococcus thermophilus in a viable count ratio of (20%-50%):(20%-50%):(20%-50%); the viable count of the composite bacterial strain is 1×10⁻⁶. 7 ~1×10 9 CFU / mL.

[0017] In one embodiment of the present invention, in step (1), the boiling sterilization is performed by boiling at 85-95°C for 20-30 minutes.

[0018] In one embodiment of the present invention, in step (1), the fermentation culture is carried out at 25-45°C until the pH of the fermentation broth is 2.50-3.50, thereby obtaining black fungus fermentation broth.

[0019] In one embodiment of the present invention, the method for preparing the black fungus pulp juice is as follows:

[0020] After crushing the black fungus into small pieces with a particle size of 1mm to 4mm, soak it in water at a ratio of 1:200 to 1:400 and cook it at 85 to 95℃ for 20 to 30 minutes. Add 2 to 6g of white sugar per 100mL during the cooking process to obtain black fungus pulp juice.

[0021] In one embodiment of the present invention, in step (3), the black fungus fermentation liquid obtained in step (1) and the black fungus pulp juice obtained in step (2) are mixed in a ratio of (1:2) to (2:1).

[0022] In one embodiment of the present invention, the preparation method of the fermented black fungus beverage for promoting bowel movement and relieving constipation is as follows:

[0023] The fermented black fungus liquid and black fungus pulp juice obtained in steps (1) and (2) are mixed in a 1:1 ratio and then compounded. The amount of xylooligosaccharide added is 1-5g / 100mL, the amount of inulin added is 1-5g / 100mL, and the amount of sodium benzoate added is 0.5g / 1000g-1g / 1000g.

[0024] In one embodiment of the present invention, the sterilization method of the fermented black fungus beverage for relieving constipation is as follows: the oral liquid obtained in step (3) is filled into a brown oral liquid bottle while it is still hot, sealed, sterilized at 85-95℃ for 20-30 minutes, and the bottle cap is tightened while it is still hot to obtain the fermented black fungus beverage for relieving constipation.

[0025] The present invention also provides a fermented black fungus beverage that promotes bowel movement and relieves constipation, prepared by the above preparation method.

[0026] Beneficial effects

[0027] Compared with the prior art, the present invention provides a method for preparing a fermented beverage stock solution of black goji berries with lactic acid bacteria, which has the following beneficial effects:

[0028] (1) This invention uses black fungus as raw material and combines it with a compound strain of bacteria (Lactobacillus bulgaricus, Streptococcus thermophilus, and Bifidobacterium longum) to prepare a fermented black fungus beverage. The prepared fermented black fungus beverage has a mild sour taste and a pleasant aroma. The fermented black fungus liquid is combined with black fungus pulp juice to increase the layers of flavor and consumer trust.

[0029] (2) The present invention extracts polysaccharide components from black fungus by enzymatic hydrolysis of compound strains. The polysaccharides have high stability during enzymatic hydrolysis. This method has the characteristics of high polysaccharide extraction rate and good activity retention.

[0030] (3) This invention utilizes black fungus polysaccharide, xylooligosaccharide and inulin to form a compound, which has a good synergistic effect in promoting bowel movement and enhancing the efficacy of the functional components of fermented black fungus beverage. While improving functionality, it can also enhance the flavor of the beverage. Attached Figure Description

[0031] Figure 1 Flowchart of the process for fermented black fungus beverage that promotes bowel movement and relieves constipation. Detailed Implementation

[0032] The technical solutions described below will be clearly and completely described with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0033] The Lactobacillus bulgaricus involved in the following examples is Lactobacillus bulgaricus HH-LB57, with a specification of 5×10 10 CFU / g, the Bifidobacterium longum is Bifidobacterium longum HH-BL18, specification 1×10 11 CFU / g, the thermophilic streptococcus is thermophilic streptococcus HH-ST08, specification 2×10 11 CFU / g were all purchased from Xi'an Jushengyuan Biotechnology Co., Ltd.

[0034] The whey protein powder used in the following examples was purchased from Zhejiang Yinuo Biotechnology Co., Ltd., and the concentrated pear juice was purchased from Guizhou Xianyuan Zhenbao Co., Ltd.

[0035] The sodium benzoate used in the following examples was purchased from Henan Wanbang Chemical Technology Co., Ltd., and the xylooligosaccharides and inulin used were purchased from Jiaowang Natural Technology Co., Ltd.

[0036] The detection methods involved in the following embodiments are as follows:

[0037] 1. Sensory evaluation

[0038] To verify the sensory effects of the fermented black fungus beverage for promoting bowel movement in this invention, 20 experienced evaluators scored the oral liquid's color, aroma, taste, and texture (0-20 points). The overall sensory score was calculated using the following formula:

[0039] Overall sensory score = color score × 30% + aroma score × 20% + taste score × 30% + texture score × 20%.

[0040] 2. Time of first black stool excretion

[0041] After animal modeling and treatment, the time to first melena excretion in mice was measured. Before the experiment, mice in each group were fasted for 16 hours, during which time they had free access to water. Each mouse in different experimental groups was administered loperamide hydrochloride (10 mL / kg) by gavage, at a dose of 0.1 mL / 10 g. 30 minutes later, all mice were administered 0.2 mL of ink by gavage, and the time of administration—initial time—was recorded for each mouse. Afterward, each mouse was placed in a separate cage, and the time to first melena excretion—the end time—was recorded. The time to first melena excretion in each group of mice according to Example 1 and Comparative Examples 1-4 was measured at 7 days, 14 days, 21 days, and 28 days.

[0042] 3. Number of fecal particles and fecal moisture content

[0043] After treatment, fresh feces excreted by each mouse within 4 hours were collected, and the wet weight of the feces was recorded. The feces were then placed in a forced-air drying oven and dried at 60°C until constant weight. The fecal moisture content was calculated using the following formula:

[0044] Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%.

[0045] Example 1: Preparation of fermented black fungus beverage and verification of its laxative function

[0046] The specific steps are as follows:

[0047] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0048] The prepared seed solutions were mixed at a ratio of 50%:20%:30% to obtain a composite strain with a viable count of 1×10⁻⁶. 8 CFU / mL.

[0049] (2) Preparation of black fungus fermentation liquid

[0050] Black fungus was pulverized to 80 mesh to obtain powder. It was soaked for 30 minutes at a material-to-water ratio of 1:80, then pulped. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. After adding 1% (v / v) of compound bacteria, it was cultured at 25℃. The fermentation endpoint was reached when the total acid (pH) reached 3.50. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0051] (3) Preparation of black fungus pulp juice

[0052] Crush dried black fungus into small particles of 4mm size, soak them in water at a ratio of 1:200, and then simmer at 85-95℃ for 20-30 minutes to sterilize. Add 2g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0053] (4) Preparation of black fungus beverage for promoting bowel movement

[0054] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a 1:1 (v / v) ratio and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 80℃ for 25 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0055] (5) Experimental method for bowel-regulating and laxative function

[0056] Thirty healthy male BALB / c mice, aged 4–6 weeks and weighing 18–22 g, were selected as experimental animals. The animal housing environment was maintained at a temperature of 23±1℃ and humidity of 40–60%, with good ventilation and a 12-hour light-dark cycle. Mice were provided with free access to water and food. A mouse constipation model was established using loperamide hydrochloride. The effects of fermented black fungus beverage on mouse constipation were investigated by measuring three indicators: the time to first black feces excretion, the number of fecal particles, and fecal water content. First, a 1 mg / mL loperamide hydrochloride solution was prepared using physiological saline. The experiment was divided into an adaptation period, an intervention period, and a modeling period, lasting a total of 25 days. The adaptation period was 0–7 days, the intervention period was 8–22 days, and the modeling period was 23–25 days.

[0057] The specific steps are as follows:

[0058] 1) After mice have adapted to growth for one week, they were randomly divided into three groups starting on day 8: blank group, model group, and oral liquid group.

[0059] Control group: From day 8 to day 22, normal saline (10 mL / kg bw) was administered by gavage twice daily; from day 23 to day 25, normal saline (10 mL / kg bw) was administered by gavage twice daily.

[0060] Model group: From day 8 to day 22, normal saline (10 mL / kg·bw) was administered by gavage twice daily; from day 23 to day 25, loperamide hydrochloride solution (10 mg / kg·bw) was administered by gavage once at 8:30 am and normal saline (10 mL / kg·bw) was administered by gavage once at 5:00 pm.

[0061] Experimental group: From day 8 to day 22, fermented black fungus beverage (10 mL / kg·bw) was administered by gavage twice daily; from day 23 to day 25, loperamide hydrochloride solution (10 mg / kg·bw) was administered by gavage once at 8:30 am and black fungus beverage (10 mL / kg·bw) was administered by gavage once at 5:00 pm.

[0062] 4) After gavage on the evening of day 24, the mice were fasted but allowed free access to water.

[0063] On day 25, mice in each group were anesthetized with isoflurane, and blood was collected from the eyeballs followed by cervical dislocation for euthanasia. Whole blood from the mice was allowed to stand at room temperature for 2 hours, then centrifuged at 3200×g for 15 minutes. The mouse serum was then aliquoted and cryopreserved at -80℃. The mice were dissected, and their intestines were segmented. A 0.5cm segment of the colon was fixed in paraformaldehyde solution, and the remaining intestinal tissue and contents were pre-frozen in liquid nitrogen and then cryopreserved at -80℃.

[0064] (6) Experimental Results

[0065] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 108.50±7.57 min, the number of stool particles was 10.13±1.93, and the stool water content was 52.40±3.98%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.2 points.

[0066] Example 2: Preparation of Fermented Black Fungus Beverage

[0067] The specific steps are as follows:

[0068] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0069] The prepared seed solutions were mixed at a ratio of 30%:50%:20% to obtain a composite strain with a viable count of 1×10⁻⁶ cells / mL. 7 CFU / mL.

[0070] (2) Preparation of black fungus fermentation liquid

[0071] Black fungus was pulverized to 120 mesh to obtain powder. It was soaked for 30 minutes and mashed at a material-to-water ratio of 1:100. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 3.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0072] (3) Preparation of black fungus pulp juice

[0073] Crush dried black fungus into small particles with a particle size of 3mm, soak them in water at a ratio of 1:200, and simmer at 85℃ for 30 minutes to sterilize. Add 3g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0074] (4) Preparation of black fungus beverage for promoting bowel movement

[0075] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a volume ratio of 2:1 (v / v) and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The bottle caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0076] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0077] (6) Experimental Results

[0078] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 111.39±5.35 min, the number of stool particles was 8.34±6.35, and the stool water content was 58.34±3.14%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.3 points.

[0079] Example 3: Preparation of Fermented Black Fungus Beverage

[0080] The specific steps are as follows:

[0081] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0082] The prepared seed solutions were mixed in a ratio of 20%:30%:50% to obtain a composite strain with a viable count of 1×10⁻⁶ cells / year. 9 CFU / mL.

[0083] (2) Preparation of black fungus fermentation liquid

[0084] Black fungus was pulverized to 180 mesh to obtain powder. It was soaked for 30 minutes and mashed at a material-to-water ratio of 1:120. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 45℃. The fermentation endpoint was reached when the total acid (pH) reached 2.50. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0085] (3) Preparation of black fungus pulp juice

[0086] Crush dried black fungus into small particles with a particle size of 2mm, soak them in water at a ratio of 1:200, and simmer at 85℃ for 30 minutes to sterilize. Add 4g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0087] (4) Preparation of black fungus beverage for promoting bowel movement

[0088] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 1:2 (v / v) and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 90℃ for 25 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0089] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0090] (6) Experimental Results

[0091] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 113.94±4.52 min, the number of stool particles was 7.39±2.49, and the stool water content was 51.34±5.35%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.3 points.

[0092] Comparative Example 1

[0093] The specific steps are as follows:

[0094] (1) Preparation of seed culture: Lactobacillus bulgaricus HH-LB57 was inoculated into MRS medium and cultured at 37℃ for 24 h. The cultured bacterial solution was centrifuged and thoroughly washed with food-grade physiological saline to prepare a bacterial suspension with a viable count of 5 × 10⁻⁶ cells / mL. 6 CFU / mL was used to obtain Lactobacillus bulgaricus seed culture;

[0095] (2) Preparation of black fungus fermentation liquid

[0096] Black fungus was pulverized to 10 mesh to obtain powder. It was soaked for 30 minutes and mashed at a material-to-water ratio of 1:150. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. After adding 2% (v / v) Lactobacillus bulgaricus seed solution, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 4.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0097] (3) Preparation of black fungus pulp juice

[0098] Crush dried black fungus into small particles with a particle size of 6mm, soak them in water at a ratio of 1:300, and simmer at 85℃ for 30 minutes to sterilize. Add 4g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0099] (4) Preparation of black fungus beverage for promoting bowel movement

[0100] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 1:3 (v / v) and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 30 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0101] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0102] (6) Experimental Results

[0103] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 121.94±4.52 min, the number of stool particles was 7.73±5.43, and the stool water content was 42.68±2.43%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 7.3 points.

[0104] Comparative Example 2

[0105] The specific steps are as follows:

[0106] (1) Preparation of seed culture: Streptococcus thermophilus HH-ST08 was inoculated into MRS medium and cultured at 37℃ for 24 h. The cultured bacterial solution was centrifuged and thoroughly washed with food-grade physiological saline to prepare a bacterial suspension with a viable count of 5 × 10⁻⁶. 6 CFU / mL was used to obtain Lactobacillus bulgaricus seed culture;

[0107] (2) Preparation of black fungus fermentation liquid

[0108] Black fungus was pulverized to 10 mesh to obtain powder. It was soaked for 30 minutes and mashed at a material-to-water ratio of 1:150. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. 2% (v / v) Streptococcus thermophilus seed solution was added and the mixture was cultured at 50℃. The fermentation endpoint was reached when the total acid (pH) reached 2.00. The clear liquid was filtered to obtain the black fungus fermentation broth.

[0109] (3) Preparation of black fungus pulp juice

[0110] Crush dried black fungus into small particles with a particle size of 6mm, soak them in water at a ratio of 1:300, and simmer at 85℃ for 30 minutes to sterilize. Add 4g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0111] (4) Preparation of black fungus beverage for promoting bowel movement

[0112] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 3:1 (v / v) and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0113] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0114] (6) Experimental Results

[0115] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 123.42±6.32 min, the number of stool particles was 6.83±0.93, and the stool water content was 49.43±7.32%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 7.2 points.

[0116] Comparative Example 3

[0117] The specific steps are as follows:

[0118] (1) Preparation of seed culture: Bifidobacterium longum HH-BL18 was inoculated into MRS medium and cultured at 37℃ for 24 h. The cultured bacterial solution was centrifuged and thoroughly washed with food-grade physiological saline to prepare a bacterial suspension with a viable count of 1×10⁻⁶. 6 CFU / mL was used to obtain a seed culture of Bifidobacterium longum;

[0119] (2) Preparation of black fungus fermentation liquid

[0120] Black fungus was pulverized to 400 mesh to obtain powder. It was soaked in water at a ratio of 1:150 for 30 minutes and then mashed. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 30 minutes to sterilize. After adding 2% (v / v) of Bifidobacterium longum seed liquid, it was cultured at 21℃. The fermentation endpoint was reached when the total acid (pH) reached 2.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0121] (3) Preparation of black fungus pulp juice

[0122] Crush dried black fungus into small particles with a particle size of 6mm, soak them in water at a ratio of 1:300, and simmer at 85℃ for 30 minutes to sterilize. Add 4g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0123] (4) Preparation of black fungus beverage for promoting bowel movement

[0124] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 1:4 (v / v) and compounded. The amount of xylooligosaccharide added was 3g / 100mL, the amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, it was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0125] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0126] (6) Experimental Results

[0127] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 120.42±5.98 min, the number of stool particles was 6.93±0.34, and the stool water content was 45.13±4.16%. Table 2 shows that the sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 7.4 points.

[0128] Example 4: Preparation of Fermented Black Fungus Beverage

[0129] The specific steps are as follows:

[0130] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0131] The prepared seed solutions were mixed in a ratio of 50%:30%:20% to obtain a composite strain with a viable count of 5 × 10⁻⁶ cells / year. 7 CFU / mL.

[0132] (2) Preparation of black fungus fermentation liquid

[0133] Black fungus was pulverized to 200 mesh to obtain powder. It was soaked for 60 minutes and mashed at a material-to-water ratio of 1:150. 0.5g / 100mL whey protein powder and 5g / 100mL concentrated pear juice were added. The mixture was boiled at 90℃ for 20 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 3.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0134] (3) Preparation of black fungus pulp juice

[0135] Crush dried black fungus into small particles with a particle size of 3mm, soak them in water at a ratio of 1:200, and simmer at 85-95℃ for 20-30 minutes to sterilize. Add 4g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0136] (4) Preparation of black fungus beverage for promoting bowel movement

[0137] The above-mentioned fermented black fungus liquid and black fungus pulp juice were mixed at a ratio of 3:2 (v / v) and compounded. The amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, the mixture was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The caps were then tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0138] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0139] (6) Experimental Results

[0140] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 86.67±5.39 min, the number of stool particles was 8.85±1.39, and the stool water content was 52.19±4.34%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.3 points.

[0141] Example 5: Preparation of Fermented Black Fungus Beverage

[0142] The specific steps are as follows:

[0143] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0144] The prepared seed solutions were mixed in a ratio of 20%:50%:30% to obtain a composite strain with a viable count of 5 × 10⁻⁶ cells / year. 8 CFU / mL.

[0145] (2) Preparation of black fungus fermentation liquid

[0146] Black fungus was pulverized to 250 mesh to obtain powder. It was soaked for 60 minutes and mashed at a material-to-water ratio of 1:150. 2g / 100mL whey protein powder and 3g / 100mL concentrated pear juice were added. The mixture was boiled at 95℃ for 20 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 3.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0147] (3) Preparation of black fungus pulp juice

[0148] Crush dried black fungus into small particles with a particle size of 2mm, soak them in water at a ratio of 1:300, and simmer at 95℃ for 20 minutes to sterilize. Add 5g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0149] (4) Preparation of black fungus beverage for promoting bowel movement

[0150] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 2:3 (v / v) and compounded. Xylooligosaccharides were added at a rate of 3g / 100mL, and sodium benzoate at a rate of 1g / 1000g. The mixture was then filled into brown oral liquid bottles while still hot, sealed, and sterilized at 95℃ for 25 minutes. The caps were tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0151] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0152] (6) Experimental Results

[0153] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 79.35±6.38 min, the number of stool particles was 8.13±2.42, and the stool water content was 54.24±3.32%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.3 points.

[0154] Example 6: Preparation of Fermented Black Fungus Beverage

[0155] The specific steps are as follows:

[0156] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0157] The prepared seed solutions were mixed in a ratio of 30%:20%:50% to obtain a composite strain with a viable count of 1×10⁻⁶ cells / year. 9 CFU / mL.

[0158] (2) Preparation of black fungus fermentation liquid

[0159] Black fungus was pulverized to 300 mesh to obtain powder. It was soaked for 60 minutes and mashed at a material-to-water ratio of 1:150. 2g / 100mL whey protein powder and 3g / 100mL concentrated pear juice were added. The mixture was boiled at 85℃ for 20 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 3.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0160] (3) Preparation of black fungus pulp juice

[0161] Crush dried black fungus into small particles with a particle size of 2mm, soak them in water at a ratio of 1:400, and simmer at 95℃ for 20 minutes to sterilize. Add 6g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0162] (4) Preparation of black fungus beverage for promoting bowel movement

[0163] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a 1:1 (v / v) ratio and compounded. The amount of inulin added was 4g / 100mL, xylooligosaccharide added was 2g / 100mL, and sodium benzoate added was 1g / 1000g. While still hot, the mixture was filled into brown oral liquid bottles, sealed, and sterilized at 90℃ for 25 minutes. The caps were then tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0164] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0165] (6) Experimental Results

[0166] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 62.58±4.83 min, the number of stool particles was 11.23±3.41, and the stool water content was 61.03±6.43%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 9.5 points.

[0167] Comparative Example 4

[0168] The specific steps are as follows:

[0169] (1) Preparation of compound bacterial strains: Lactobacillus bulgaricus HH-LB57, Bifidobacterium longum HH-BL18 and Streptococcus thermophilus HH-ST08 were inoculated into MRS medium and cultured at 37℃ for 24 h. The bacterial suspensions were centrifuged and thoroughly washed with food-grade physiological saline to prepare bacterial suspensions, and Lactobacillus bulgaricus seed liquid, Bifidobacterium longum seed liquid and Streptococcus thermophilus seed liquid were prepared respectively.

[0170] The prepared seed solutions were mixed in a ratio of 50%:20%:30% to obtain a composite strain with a viable count of 1×10⁻⁶ cells / year. 10 CFU / mL.

[0171] (2) Preparation of black fungus fermentation liquid

[0172] Black fungus was pulverized to 20 mesh to obtain powder. It was soaked for 60 minutes and mashed at a material-to-water ratio of 1:100. 2g / 100mL whey protein powder and 3g / 100mL concentrated pear juice were added. The mixture was boiled at 95℃ for 20 minutes to sterilize. After adding 2% (v / v) of compound bacteria, it was cultured at 35℃. The fermentation endpoint was reached when the total acid (pH) reached 3.00. The clear liquid was filtered to obtain the black fungus fermentation liquid.

[0173] (3) Preparation of black fungus pulp juice

[0174] Crush dried black fungus into small particles with a particle size of 7mm, soak them in water at a ratio of 1:200, and simmer at 85℃ for 20 minutes to sterilize. Add 2g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0175] (4) Preparation of black fungus beverage for promoting bowel movement

[0176] The above-mentioned fermented black fungus liquid and black fungus pulp juice were mixed at a ratio of 1:5 (v / v) and compounded. No inulin or xylooligosaccharides were added, and the sodium benzoate content was 1g / 1000g. The mixture was filled into brown oral liquid bottles while hot, sealed, and sterilized at 85℃ for 25 minutes. The caps were tightened while hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0177] (5) The experimental method for the bowel-regulating and laxative function is the same as in Example 1;

[0178] (6) Experimental Results

[0179] The results of the bowel-regulating and laxative effect experiment are shown in Tables 11 and 2. The results showed that the time to expel the first black stool was 133.43±23.42 min, the number of stool particles was 4.39±1.93, and the stool water content was 47.24±3.42%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 7.5 points.

[0180] Comparative Example 5

[0181] The specific steps are as follows:

[0182] (1) Preparation of black fungus extract

[0183] Black fungus was pulverized to 600 mesh to obtain powder. It was soaked for 60 minutes and mashed into a paste at a material-to-water ratio of 1:100. 2g / 100mL whey protein powder and 3g / 100mL concentrated pear juice were added. The mixture was boiled at 95℃ for 20 minutes to sterilize. The clear liquid was filtered and obtained without fermentation to obtain black fungus extract.

[0184] (2) Preparation of black fungus pulp juice

[0185] Crush dried black fungus into small particles with a particle size of 7mm, soak them in water at a ratio of 1:200, and simmer at 85℃ for 20 minutes to sterilize. Add 2g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0186] (3) Preparation of black fungus beverage for promoting bowel movement

[0187] The above-mentioned fermented black fungus liquid and black fungus pulp juice were mixed at a ratio of 5:1 (v / v) and compounded. The amount of inulin added was 3g / 100mL, and the amount of sodium benzoate added was 1g / 1000g. While still hot, the mixture was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The caps were then tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0188] (4) The method for the test of bowel-regulating and laxative function is the same as in Example 1;

[0189] (5) Experimental Results

[0190] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 122.34±15.82 min, the number of stool particles was 5.34±0.42, and the stool water content was 44.42±1.38%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 8.2 points.

[0191] Comparative Example 6

[0192] The specific steps are as follows:

[0193] (1) Preparation of black fungus extract

[0194] Black fungus was pulverized to 30 mesh to obtain powder. It was soaked in water at a ratio of 1:100 for 60 minutes and then mashed. 2g / 100mL whey protein powder and 3g / 100mL concentrated pear juice were added. The mixture was boiled at 95℃ for 20 minutes to sterilize. The clear liquid was filtered and obtained without fermentation to obtain black fungus extract.

[0195] (2) Preparation of black fungus pulp juice

[0196] Crush dried black fungus into small particles of 5mm size, soak them in water at a ratio of 1:200, and simmer at 85℃ for 20 minutes to sterilize. Add 2g / 100mL of white sugar during the simmering process to obtain black fungus pulp juice.

[0197] (3) Preparation of black fungus beverage for promoting bowel movement

[0198] The above-mentioned fermented black fungus broth and black fungus pulp juice were mixed at a ratio of 1:1 (v / v) and compounded. Xylooligosaccharide was added at a rate of 3g / 100mL, and sodium benzoate at a rate of 1g / 1000g. While still hot, the mixture was filled into brown oral liquid bottles, sealed, and sterilized at 85℃ for 25 minutes. The caps were then tightened while still hot to obtain the fermented black fungus beverage that promotes bowel movement and relieves constipation.

[0199] (4) The method for the test of bowel-regulating and laxative function is the same as in Example 1;

[0200] (5) Experimental Results

[0201] The results of the bowel-regulating and laxative effect experiment are shown in Tables 1 and 2. The results showed that the time to expel the first black stool was 119.31±7.73 min, the number of stool particles was 5.82±1.92, and the stool water content was 48.45±1.31%. The sensory evaluation score of the fermented black fungus beverage for bowel regulation and laxative effect was 8.0 points.

[0202] Example 7: Experimental Results

[0203] The specific results of the above embodiments and comparative examples are shown in Tables 1 and 2:

[0204] Table 1: Results of the experiment on bowel-regulating and laxative functions

[0205]

[0206]

[0207] Table 2: Sensory evaluation results of black fungus oral liquid

[0208]

[0209] The comparison results of Examples 1, 2, and 3 with Comparative Examples 1, 2, and 3 show that fermentation of black fungus extract using a compound bacteria has better sensory effects and also improves bowel function. The compound bacteria (20-50% Lactobacillus bulgaricus, 20-50% Bifidobacterium longum, and 20-50% Streptococcus thermophilus), fermentation temperature of 25-45℃, and fermentation acidity of 2.50-3.50 significantly improved the sensory score of the oral liquid, far exceeding that of the single-strain or comparative examples with fermentation temperatures and acidities outside the above ranges. In particular, it improved the aroma and taste scores of the oral liquid; the aroma and taste of Example 3 were 23% and 11% higher than those of Comparative Example 1, respectively.

[0210] The overall sensory scores of Examples 1, 2, and 3 were all above 90, with a sensory effect improvement of 25.17% to 29.89%. This indicates that the fermented black fungus beverage produced using the production process of this invention has the characteristics of excellent taste and strong function.

[0211] The comparative results of Examples 4, 5, and 6 with Comparative Examples 4, 5, and 6 show that the oral liquid obtained by combining fermented black fungus polysaccharide solution with inulin and xylooligosaccharide, in Example 6, significantly enhances the laxative effect compared to Comparative Example 4, which uses only black fungus polysaccharide. Example 6 shortened the time to first stool excretion in experimental mice by 53.10%, increased stool volume by 155.81%, and increased stool water content by 29.19%, demonstrating a significant improvement in laxative function. This effect also far exceeds the combined effect of Comparative Example 5 (inulin alone) and Comparative Example 6 (xylooligosaccharide alone). This indicates that black fungus polysaccharide, inulin, and xylooligosaccharide have a synergistic effect in promoting laxative function.

[0212] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for preparing a fermented Auricularia auricula-judae beverage having a bowel-lubricating and bowel-moving function, characterized by, The method comprises the following steps: (1) Preparation of Auricularia auricular fermenting liquid The black fungus is crushed, then soaked, beaten, added with whey protein powder and concentrated pear juice, boiled and sterilized, cooled, added with a compound strain, fermented and cultured, filtered to obtain a black fungus fermentation liquor; the compound strain is obtained by mixing Lactobacillus bulgaricus, Bifidobacterium longum and Streptococcus thermophilus in a ratio of (20%-50%):(20%-50%):(20%-50%) of live bacteria number, and the inoculation amount of the compound strain is 1%-3%; the live bacteria number of the compound strain is: 1x10 7 ~1x10 9 CFU / mL. (2) Preparation of Auricularia auricular pulp juice The Auricularia auricular is crushed into small particles with a particle size of 1 mm-4 mm, soaked in water, boiled after adding sugar, and Auricularia auricular pulp juice is obtained; (3) Preparation of Auricularia auricular beverage with the function of moistening intestines and defecation The Auricularia auricular fermenting liquid obtained in step (1) and the Auricularia auricular pulp juice obtained in step (2) are mixed in a ratio of (1:2) to (2:1), oligosaccharides, inulin, and sodium benzoate are added, and after filling, sealing, and sterilization, the fermented Auricularia auricular beverage is obtained. The addition amount of oligosaccharides is 1 g / 100 mL-5 g / 100 mL, the addition amount of inulin is 1 g / 100 mL-5 g / 100 mL, and the addition amount of sodium benzoate is 0.5 g / 1000 g-1 g / 1000 g.

2. The production method according to claim 1, characterized by, In step (1), the Auricularia auricular is crushed to 80-300 mesh, soaked in water in a ratio of (1:80) to (1:150), and soaked for 20-60 min. After beating, 0.5 g / 100 mL-2 g / 100 mL of whey protein and 1 g / 100 mL-7 g / 100 mL of concentrated pear juice are added.

3. The preparation method according to claim 1, characterized in that, In step (1), the boiling sterilization is: boiling at 85-95℃ for 20-30 min.

4. The production method according to claim 3, characterized by, In step (1), the fermentation culture is carried out at 25-45℃, and the pH of the fermentation liquid is 2.50-3.50 after culture, to obtain Auricularia auricular fermenting liquid.

5. The preparation method according to claim 4, characterized in that, In step (2), the Auricularia auricular is crushed into small particles with a particle size of 1 mm-4 mm, soaked in water in a ratio of (1:200) to (1:400), and boiled at 85-95℃ for 20-30 min. 2 g / 100 mL-6 g / 100 mL of white granulated sugar is added during boiling to obtain Auricularia auricular pulp juice.

6. The method of claim 1, wherein, In step (3), the sterilization conditions are 85-95℃, 20-30 min.

7. The fermented Auricularia auricular beverage with the function of moistening intestines and defecation prepared by the preparation method of any one of claims 1-6.

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