Glutinous rice banana fermentation liquor as well as preparation method and application thereof

Abstract

The present invention discloses a glutinous rice banana fermentation broth and a preparation method and application thereof, and belongs to the field of fermented food processing, the glutinous rice banana fermentation broth is prepared by taking glutinous rice banana as a raw material and adopting a compound enzyme and a microbial agent through the processes of peeling, pulping, constant temperature enzymolysis, ingredient fermentation and the like, the crude polysaccharide content of the glutinous rice banana fermentation broth is 1.34 g / mL, the total acid content is 137.95 g / L, and the total acid content is 17.95 g / L. The composition contains seven organic acids: oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid, maleic acid and succinic acid. Animal experiments prove that the glutinous rice banana fermentation liquor has the effect of promoting small intestine peristalsis, increasing the humidity of excrement, obviously improving the ink advancing rate of intestinal tracts, shortening the first-grain black excrement discharging time and relieving mouse constipation. By regulating the secretion of water and electrolyte in the intestinal tract of a constipation mouse and the peristalsis of the small intestine, the loperamide hydrochloride-induced constipation model mouse has the effects of relaxing bowel and protecting the intestinal tract.

Description

Technical Field

[0001] The present invention belongs to the technical field of fermented foods, and particularly relates to a fermented solution of Musa acuminata Colla and a preparation method and application thereof. Background Art

[0002] With the increasing refinement of people's diet in modern society, it has led to overnutrition, making constipation a very common disease. The main characteristics of constipation are difficult defecation, hard stools, and a reduced frequency of bowel movements. When constipated, many patients will choose to use laxatives such as aloe vera, senna leaves, and rhubarb, most of which contain anthraquinone chemical substances. However, data shows that the abuse of laxatives containing anthraquinones may cause patients to be dependent on anthraquinone laxatives and induce colon lesions, thereby increasing the risk of deformation into colorectal cancer. Since natural products themselves have the characteristics of being green and safe, it is urgent and necessary to find natural products that can promote intestinal peristalsis, produce water and electrolytes, and increase the water content of feces. Compared with traditional anthraquinone laxatives, the green and safe characteristics of Musa acuminata Colla more meet the modern people's pursuit of health. As a natural and green food in nature, Musa acuminata Colla is often regarded as an important part of a healthy diet in people's daily diet due to its rich nutritional components, and the bioactive compounds contained in Musa acuminata Colla may have the potential to promote intestinal health.

[0003] Glutinous banana, also known as powder banana and beauty banana, belongs to the group of Musa ABB Pisangawak in the genus Musa (MusaL.) of the family Musaceae (Musaceae), and is a native plant in Southeast Asia. Glutinous banana has a unique taste, is delicious, rich in nutrition, and affordable, and is deeply loved by consumers. Glutinous banana contains rich nutrients such as protein, riboflavin, fat, calcium, phosphorus, iron, and carbohydrates, and also contains bioactive compounds such as phenols, carotenoids, and phytosterols. In addition, glutinous banana is rich in potassium, which helps to maintain the electrolyte balance of the human body and prevent the occurrence of hypokalemia; at the same time, glutinous banana is also rich in resistant starch, which has the functions of promoting gastrointestinal peristalsis and laxative. It has high nutritional value and has always been an indispensable part of people's healthy diet. Despite such high nutritional value of glutinous banana, glutinous banana belongs to climacteric fruits. Glutinous banana is prone to mechanical damage during the ripening process, resulting in varying degrees of oxidative deterioration during transportation and storage, affecting the nutrients contained in glutinous banana and being inconvenient for long-term transportation and storage. At present, the by-products processed from bananas mainly include banana compound juice beverages, banana yogurt, banana vinegar, banana wine, and banana jam. Therefore, in order to solve the challenges faced by glutinous banana during long-term transportation and storage, developing related by-products of glutinous banana has become an effective means. This can not only reduce the loss of nutrients, but also expand the use value of glutinous banana and further explore its application in the food industry. Therefore, it is urgent to seek a new method to effectively expand the utilization ways of glutinous banana. Summary of the Invention

[0004] The purpose of this part is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments. Simplifications or omissions may be made in this part, as well as in the abstract and title of the specification of this application, to avoid obscuring the purpose of this part, the abstract, and the title, and such simplifications or omissions shall not be used to limit the scope of the present invention.

[0005] In view of the above and / or problems existing in the prior art, the present invention is proposed.

[0006] Therefore, the purpose of the present invention is to overcome the deficiencies in the prior art and provide a glutinous banana fermentation broth.

[0007] To solve the above technical problems, the present invention provides the following technical solution: A glutinous banana fermentation broth, characterized in that: the content of crude polysaccharide in the glutinous banana fermentation broth is 1.34 g / mL, the total acid content is 137.95 g / L, and it contains 7 organic acids including oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid, maleic acid, and succinic acid.

[0008] Another purpose of the present invention is to overcome the deficiencies in the prior art and provide a preparation method of a glutinous banana fermentation broth, characterized in that: it includes,

[0009] Pulp peeling and pulping: Take fresh, intact, green-skinned plantains with a plumpness of 75% - 80%, peel and crush them to obtain plantain pulp;

[0010] Constant temperature enzymatic hydrolysis: Sterilize the plantain pulp at high temperature, and after cooling, add glucoamylase, cellulase, pectinase, and lipase, and perform enzymatic hydrolysis to obtain an enzymatic hydrolysis solution of plantain pulp;

[0011] Ingredient fermentation: Mix the enzymatic hydrolysis solution of plantain pulp, purified water, and granulated sugar, add probiotic liquid, control the fermentation temperature at 15 - 25 °C, and perform aerobic fermentation for 10 - 30 days. Stir once every 3 days during the fermentation process until the solid content is 7% - 8% and the pH value is about 4, then take the supernatant and filter it to obtain the plantain fermentation liquid.

[0012] As a preferred embodiment of the preparation method of the present invention, wherein: for the constant temperature enzymatic hydrolysis, glucoamylase 0.05 - 0.5%, cellulase 0.05 - 0.5%, pectinase 0.05 - 0.5%, and lipase 0.01 - 0.05% are added according to the total weight of the pulp.

[0013] As a preferred embodiment of the preparation method of the present invention, wherein: when mixing the enzymatic hydrolysis solution of plantain pulp, purified water, and granulated sugar, by mass fraction, the mass ratio of the enzymatic hydrolysis solution of plantain pulp, purified water, and granulated sugar is 5 - 10:20 - 35:2 - 8.

[0014] As a preferred embodiment of the preparation method of the present invention, wherein: for the addition of probiotic liquid, the preparation method of the probiotic liquid is using black tea juice as the basic culture medium, adding granulated sugar according to 8 - 10% of the weight of the black tea juice, and simultaneously adding yeast powder, lactobacillus powder, and acetic acid bacteria powder, and culturing under aerobic conditions for 10 - 15 hours.

[0015] As a preferred embodiment of the preparation method of the present invention, wherein: the viable count of yeast in the probiotic liquid is 1.0×10 5 CFU / mL, the viable count of lactobacillus is 1.0×10 6 CFU / mL, and the viable count of acetic acid bacteria is 1.0×10 4 CFU / mL.

[0016] Another object of the present invention is to overcome the deficiencies in the prior art and provide an application of a plantain fermentation liquid, characterized in that: the application of the plantain fermentation liquid in preparing a health food or drug with the function of promoting defecation and relieving constipation.

[0017] As a preferred embodiment of the application of the present invention, the fermented liquid of Musa itinerans Cheesman can improve the intestinal peristalsis and defecation function of constipated mice by reducing the vasoactive intestinal peptide in the serum and increasing the gastrin and motilin.

[0018] As a preferred embodiment of the application of the present invention, the fermented liquid of Musa itinerans Cheesman has the functions of moistening the intestine and relieving constipation and protecting the intestine for the constipation model mice induced by loperamide hydrochloride by regulating the secretion of water and electrolytes in the intestine of constipated mice and the peristalsis of the small intestine.

[0019] Another object of the present invention is to overcome the deficiencies in the prior art and provide a drink with the function of moistening the intestine and relieving constipation, which is characterized in that the drink uses the fermented liquid of Musa itinerans Cheesman described in claim 1 as an active ingredient.

[0020] Beneficial effects of the present invention:

[0021] (1) The present invention uses Musa itinerans Cheesman as a raw material and is prepared by processes such as peeling and pulping, constant-temperature enzymatic hydrolysis, and ingredient fermentation with a composite enzyme and a microbial inoculant. The crude polysaccharide content of the fermented liquid of Musa itinerans Cheesman is 1.34 g / mL, the total acid content is 137.95 g / L, and it contains 7 organic acids, namely oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid, maleic acid, and succinic acid.

[0022] (2) The fermented liquid of Musa itinerans Cheesman described in the present invention has the effect of promoting small intestine peristalsis, increasing the moisture of feces, significantly increasing the intestinal ink propulsion rate, shortening the time of the first black feces excretion, and relieving constipation in mice. By regulating the secretion of water and electrolytes in the intestine of constipated mice and the peristalsis of the small intestine, it has the functions of moistening the intestine and relieving constipation and protecting the intestine for the constipation model mice induced by loperamide hydrochloride. Description of the drawings

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings. Among them:

[0024] Figure 1 Shows the effect of the fermented liquid of Musa itinerans Cheesman on the body weight of mice;

[0025] Figure 2 Shows the effect of the fermented liquid of Musa itinerans Cheesman on the defecation parameters of mice;

[0026] Figure 3 Shows the effect of the fermented liquid of Musa itinerans Cheesman on the small intestine of mice;

[0027] Figure 4Effect of fermented liquid of Musa itinerans Cheesman on liver coefficient and kidney coefficient of mice;

[0028] Figure 5 Effect of fermented liquid of Musa itinerans Cheesman on serum level of mice;

[0029] Figure 6 Effect of fermented liquid of Musa itinerans Cheesman on the tissue structure of jejunal villi of mice (HE, 50×). Specific implementation mode

[0030] In order to make the above objects, features and advantages of the present invention more obvious and understandable, the specific implementation mode of the present invention will be described in detail below in combination with the embodiments of the specification.

[0031] In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Those skilled in the art can make similar promotions without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0032] Secondly, the so-called "one embodiment" or "embodiment" herein refers to a specific feature, structure or characteristic that can be included in at least one implementation mode of the present invention. The "in one embodiment" appearing in different places in this specification does not all refer to the same embodiment, nor is it a separate or alternative embodiment that excludes other embodiments.

[0033] Materials used in the embodiments of the present invention: Probiotic liquid: self-made by Guizhou Natural Product Research Center (the ratio of yeast: acetic acid bacteria: lactic acid bacteria is 1:1:1); Loperamide Hydrochloride Capsules: Janssen Pharmaceutical Co., Ltd.; Lipase (50,000 U / g), Cellulase (20,000 U / g), Glucoamylase (50,000 U / g), Pectinase (30,000 U / g): Nanning Pangbo Bioengineering Co., Ltd.; Activated carbon (food grade): Jinling Carbon Industry; Mosapride Citrate: Fujian Haixi New Drug Innovation Co., Ltd.; Mouse Substance P (SP) ELISA Kit, Gastrin (GAS) ELISA Kit, Tumour necrosis factor-alpha (TNF-α) ELISA Kit, Vasoactive Intestinal Peptide (VIP) ELISA Kit, Motilin (MTL) ELISA Kit: Quanzhou Ruixin Biotechnology Co., Ltd.; Organic acid standard products (Sinopharm Chemical Reagent Co., Ltd.); Sodium hydroxide (Chengdu Jinshan Chemical Reagent Co., Ltd.); Phenolphthalein (Tianjin Kemiou Chemical Reagent Co., Ltd.).

[0034] Instruments used in the embodiments of the present invention: burette (10.00 mL); U3000 high performance liquid chromatography (Thermo Fisher Scientific); UV1800 ultraviolet spectrophotometer; 5430r high-speed refrigerated centrifuge, Eppendorf, Germany; YH-A20002 electronic balance, Wuxin Weighing Co., Ltd.; BSA323S precision electronic balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.; SKG-PB-936 beater, Shanghai Daruibao Food Machinery Co., Ltd., China; Epoch microplate reader, BioTeK; RM2016 microtome, Shanghai Leica Instruments Co., Ltd.

[0035] The probiotic liquid used in the embodiments of the present invention is based on black tea juice as the basic culture medium, adding granulated sugar at 8% of the weight of the black tea juice, and at the same time adding yeast powder, lactobacillus powder, and acetic acid bacteria powder and culturing under aerobic conditions for 15 hours. Among them, the viable count of yeast in the probiotic liquid is 1.0×10 5 CFU / mL, the viable count of lactobacillus is 1.0×10 6 CFU / mL, and the viable count of acetic acid bacteria is 1.0×10 4 CFU / mL.

[0036] Example 1

[0037] This example provides a method for preparing a fermented liquid of Musa acuminata Colla cv. Cavendish, comprising the following steps:

[0038] (1) Pulp peeling and pulping: Take fresh, intact-shaped, green-skinned Musa acuminata Colla cv. Cavendish with a plumpness of 75%, peel it, and use a beater to crush it to obtain Musa acuminata Colla cv. Cavendish pulp.

[0039] (2) Constant temperature enzymatic hydrolysis: Place the Musa acuminata Colla cv. Cavendish pulp in a sterilizer and sterilize it at 120°C for 10 minutes, then cool it to 40°C. Add 0.05% saccharifying enzyme, 0.5% cellulase, 0.05% pectinase, and 0.05% lipase according to the total weight of the pulp, and carry out enzymatic hydrolysis at 40°C under insulation conditions for 1.5 h to obtain an enzymolysis solution of Musa acuminata Colla cv. Cavendish pulp.

[0040] (3) Ingredient fermentation: Mix the obtained enzymolysis solution of Musa acuminata Colla cv. Cavendish pulp with 10 parts of raw material enzymolysis solution of Musa acuminata Colla cv. Cavendish pulp, 20 parts of purified water, and 8 parts of granulated sugar. Add 10% of the probiotic liquid according to the total weight of the mixture, control the fermentation temperature at 25°C, carry out aerobic fermentation for 10 days, stir once every 3 days during the fermentation process, and filter the supernatant until the solid content is 8% and the pH value is about 4 to obtain the fermented liquid of Musa acuminata Colla cv. Cavendish.

[0041] Example 2

[0042] This example provides a method for preparing a fermented liquid of Musa acuminata Colla cv. Cavendish, comprising the following steps:

[0043] (1) Pulp peeling and pulping: Take fresh, intact, green-skinned plantains with a plumpness of 80%, peel them, and use a pulping machine to crush them to obtain plantain pulp;

[0044] (2) Constant-temperature enzymatic hydrolysis: Place the plantain pulp in a sterilization pot and sterilize it at 100°C for 15 minutes, then cool it to 30°C. Add 0.1% saccharifying enzyme, 0.1% cellulase, 0.1% pectinase, and 0.015% lipase according to the total weight of the pulp, and carry out enzymatic hydrolysis for 1.5 h under the condition of keeping warm at 40°C to obtain the enzymatically hydrolyzed solution of plantain pulp;

[0045] (3) Ingredient fermentation: Mix the obtained enzymatically hydrolyzed solution of plantain pulp with 9 parts of raw material enzymatically hydrolyzed solution of plantain pulp, 27 parts of purified water, and 4 parts of granulated sugar. Add probiotic liquid according to 8% of the total weight of the mixture, control the fermentation temperature at 20°C, and carry out aerobic fermentation for 20 days. Stir once every 3 days during the fermentation process until the solid content is 7% - 8% and the pH value is about 4, then take the supernatant and filter it to obtain the plantain fermentation liquid.

[0046] Example 3

[0047] This example provides a method for preparing a plantain fermentation liquid, including the following steps:

[0048] (1) Pulp peeling and pulping: Take fresh, intact, green-skinned plantains with a plumpness of 78%, peel them, and use a pulping machine to crush them to obtain plantain pulp;

[0049] (2) Constant-temperature enzymatic hydrolysis: Place the plantain pulp in a sterilization pot and sterilize it at 110°C for 12 minutes, then cool it to 35°C. Add 0.3% saccharifying enzyme, 0.3% cellulase, 0.3% pectinase, and 0.05% lipase according to the total weight of the pulp, and carry out enzymatic hydrolysis for 1.5 h under the condition of keeping warm at 40°C to obtain the enzymatically hydrolyzed solution of plantain pulp;

[0050] (3) Ingredient fermentation: Mix the obtained enzymatically hydrolyzed solution of plantain pulp with 10 parts of raw material enzymatically hydrolyzed solution of plantain pulp, 20 parts of purified water, and 8 parts of granulated sugar. Add probiotic liquid according to 10% of the total weight of the mixture, control the fermentation temperature at 15°C, and carry out aerobic fermentation for 30 days. Stir once every 3 days during the fermentation process until the solid content is 8% and the pH value is about 4, then take the supernatant and filter it to obtain the plantain fermentation liquid.

[0051] In order to further evaluate the effect of the plantain fermentation liquid of the present invention on promoting defecation, the following tests were carried out using the plantain fermentation liquid prepared in Example 2 as the test object.

[0052] (1) Component analysis of the plantain fermentation liquid

[0053] The method for determining the crude polysaccharide content of the fermented solution of Glucoma bananas refers to SN / T 4260-2015. The method for determining the total acid content refers to GB12456-2021. The fermented solution of Glucoma bananas was diluted 10 times and calculated based on tartaric acid. The organic acid content was determined by high performance liquid chromatography.

[0054] Sample preparation: Weigh about 1 g of the homogeneous sample (accurate to 0.0001 g), dissolve and make up the volume to 50 mL with deionized water solvent, and filter through a 0.45 μm filter membrane for injection; Instrument conditions: Chromatographic column: syncronis C18, Dim 250 mm×4.6 mm, 5 μm, injection volume: 20 μL, column temperature: 30 °C, flow rate: 0.5 mL / min, mobile phase: water (0.05 mol / L potassium dihydrogen phosphate adjusted to pH = 2.68 with phosphoric acid) = 100, detect the absorption peak under ultraviolet light at a wavelength of 210 nm.

[0055] Determination of the crude polysaccharide content of the fermented solution of Glucoma bananas: The standard curve equation of glucose is Y = 9.05X + 0.0045 (R2 is 0.9991). The sample of the fermented solution of Glucoma bananas was diluted 2000 times, 1.0 mL was taken, and the measured crude polysaccharide content was 1.34 g / mL.

[0056] Determination of the total acid content of the fermented solution of Glucoma bananas: The total acid content of the fermented solution of Glucoma bananas was 137.95 g / L.

[0057] Determination of the organic acid content of the fermented solution of Glucoma bananas: The components and contents of organic acids in fruits are important compositional factors of fruits. The common organic acids in fruits mainly include L-malic acid, citric acid, tartaric acid, oxalic acid, lactic acid, acetic acid, etc. In this invention, standard curves were drawn with 11 organic acids including oxalic acid, tartaric acid, formic acid, malic acid, lactic acid, acetic acid, maleic acid, citric acid, fumaric acid, succinic acid and propionic acid; Seven organic acids were detected in the fermented solution of Glucoma bananas, and the acetic acid content was the highest.

[0058] Table 1 Organic acids and contents in the fermented solution of Glucoma bananas

[0059]

[0060] (2) Grouping of experimental mice and administration doses

[0061] SPF-grade male ICR mice (18 - 22 g) were used as the research subjects. They were housed in an animal laboratory with a humidity of 60% ± 10%, a temperature of 22 ± 2°C, and a light-dark cycle of 12 h. After one week of adaptive feeding during which they had free access to water and food, the animal experiment conditions were approved by the Animal Experiment Ethics Committee of Guizhou Medical University and met the established ethical requirements. After the adaptive feeding, the mice were randomly divided into 12 groups, and then these 12 groups were randomly assigned to two experimental groups, A and B. Each experimental group had 6 subgroups (N = 8), namely the Normal Group (NG), the Constipation Group (CG), the Mosapride citrate Group (MoC), the Glutinous Rice Banana Enzyme Low Dose Group (GRBE-L), the Glutinous Rice Banana Enzyme Medium Dose Group (GRBE-M), and the Glutinous Rice Banana Enzyme High Dose Group (GRBE-H).

[0062] Among them, the positive drug group was intragastrically administered mosapride citrate (2.5 mg / kg·bw) according to the mouse body weight, which was obtained by converting the human-mouse dose. GRBE-H was given the undiluted glutinous rice banana enzyme solution, GRBE-M was given the glutinous rice banana enzyme solution diluted 2 times, and GRBE-L was given the glutinous rice banana enzyme solution diluted 4 times (obtained from preliminary experiments). They were administered according to the body weight of each group of mice (see Table 1 for details). The NG group and the CG group were intragastrically administered distilled water (10 mL / kg·bw) according to the mouse body weight. During the administration period, the body weights of the mice in each group were weighed at the same time point every 3 days and recorded.

[0063] Table 2 Gastric gavage doses of mice in each group

[0064]

[0065] Compared with the CG group, the mice in the NG, MoC, GRBE-H, GRBE-M, and GRBE-L groups had smooth and shiny hair and good mental state. By Figure 1It can be seen that the change trend of the body weight of mice in each group from 0 to 8 days after drug administration. Compared with the NG group, the body weight gain of mice in the GRBE-H, GRBE-M, and GRBE-L groups was relatively slow during the drug administration period. The body weight of mice in each group showed an increasing trend during the period of 0 to 4 days. During the period of 4 to 8 days, the body weight of mice in the NG and CG groups continued to increase, but the weight gain was relatively gentle. During this period, the body weight of mice in the MoC, GRBE-L, GRBE-M, and GRBE-H groups showed a downward trend, and the downward trend of the body weight of mice in the GRBE-M and GRBE-H groups was more obvious. In addition, during the gavage period, it was found that the feces of mice in the GRBE-L, GRBE-M, and GRBE-H groups had different degrees of moisture with the increase of the drug dosage. The feces of mice in the GRBE-M and GRBE-H groups were more moist than those of mice in the GRBE-L group. This may be due to the relatively faster intestinal peristalsis and slightly decreased body weight of mice in the GRBE-M and GRBE-H groups (Note: compared with the NG group, * indicates P < 0.05, * indicates P < 0.01, ** indicates P < 0.001).

[0066] (3) Laxative experiment

[0067] Establishment of the mouse constipation model: After gavage with the corresponding drugs according to the doses of mice in each group for 7 days, on the 7th day, the mice in each group were fasted for 16 h (free drinking water); the experiment was carried out at 9:00 am on the 8th day. Except for the NG group given an equal amount of distilled water, the other groups were given loperamide hydrochloride (LH) 10 mg / kg·bw to establish the mouse constipation model.

[0068] Index evaluation method: 0.5 h after giving LH, except for the NG group and the CG group given 5% ink without the test sample according to the body weight of mice by gavage, the mice in the other groups were given 5% ink containing the corresponding test sample according to their body weights by gavage. The mice in the A experimental group were individually caged (a piece of absorbent paper was placed in the mouse cage) after gavage with the corresponding ink, without water and food restriction. At the same time, all observers started timing with the same type of timer. In the next 5 h, each observer recorded the time of the first black stool, the weight of the black stool, and the number of black stools excreted by each mouse (weighed using the same electronic balance), and recorded the fecal traits of the mice in each group of the A experimental group; the moisture content of the black stools of each group of mice was calculated according to formula (1) after drying at 60 °C.

[0069]

[0070] From Figure 2It can be seen that the time for the first black stool excretion in the CG group of mice was significantly longer than that in the NG group, and the number of black stools, the wet weight of black stools, and the water content of black stools excreted by the GC group of mice within 5 h were significantly lower than those in the blank group. This indicates that the constipation model of mice was successfully established, and the method of using LH to establish the constipation model of mice is reasonable and feasible. In addition, after continuous 7-day pre-intervention with various doses of GRBE, the time for the first black stool excretion of each group of constipated mice was shortened to varying degrees compared with the CG group, and the number of black stools, the wet weight of black stools, and the water content of black stools excreted within 5 h all increased to varying degrees compared with the CG group, showing a dose relationship.

[0071] In the first black stool excretion experiment, there were significant differences between the GRBE-M and GRBE-H groups and the CG group (P < 0.05); while there was no significant difference between the GRBE-L group and the CG group. The number of black stools, the wet weight of black stools, and the water content of black stools excreted by the GRBE-H group of mice within 5 h had better improvement effects compared with the CG group, with significant differences (P < 0.001); there was a significant difference in the number of black stools excreted by the GRBE-M group of mice within 5 h compared with the CG group (P < 0.05); there were significant differences in the wet weight of black stools and the water content of black stools excreted by the GRBE-M group of mice within 5 h compared with the CG group (P < 0.01); while there were no significant differences in the number of black stools, the wet weight of black stools, and the water content of black stools excreted by the GRBE-L group of mice within 5 h compared with the CG group. Thus, it can be seen that GRBE has the effect of moistening the intestines and promoting defecation, helps mice defecate, reduces the phenomenon of hard feces, and the effect of GRBE-H is the most significant, and each dose shows a dose dependence. (Note: A is the time for the first black stool excretion; B is the number of black stools; C is the weight of black stools; D is the water content of black stools. Compared with the NG group, # indicates P < 0.05, ## indicates P < 0.01, indicates P < 0.001; compared with the CG group, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001, the same below)

[0072] (4) Small intestine motility experiment

[0073] Establishment of a mouse small intestine peristalsis inhibition model: After intragastric administration of the corresponding drugs to mice in each group for 7 days according to the dose, on the 7th day, the mice in each group were fasted for 16 h (with free access to water); on the morning of the 8th day at 9:00, the experiment was carried out. Except for the NG group which was given an equal amount of distilled water, the other groups were given LH at 5 mg / kg·bw to establish a mouse intestinal peristalsis inhibition model. Index evaluation method: 0.5 h after administration of LH, except for the NG group and the CG group which were given 5% ink without the test sample by intragastric administration according to the body weight of the mice, the other groups of mice were given 5% ink containing the corresponding test sample by intragastric administration according to their body weights. 25 min later, the mice were immediately sacrificed by cervical dislocation. The mice were dissected and the small intestine from the lower end of the stomach (pylorus) to the upper end of the cecum was taken, gently stretched into a straight line and placed on a white board. The length from the pylorus to the ink front was the ink propulsion length, and the length of the intestinal tube was the total length of the small intestine, and the data were recorded. The ink propulsion rate was calculated according to formula (2).

[0074]

[0075] As Figure 3 shown, the results of the small intestine length and ink propulsion rate of mice in each group are shown in Fig. 2A. There was no significant difference in the small intestine length among mice in each group (P > 0.05); as shown in Figure 2 B, the ink propulsion rate of the CG group was significantly lower than that of the NG group, indicating that this modeling method was feasible and the modeling was successful. It can be seen from the figure that compared with the CG group, the ink propulsion rates of the GRBE-M and GRBE-H groups of mice were significantly increased (P < 0.001), and the ink propulsion rate of the GRBE-M group was close to that of the NG group, and the ink propulsion rate of the GRBE-H group was close to that of the MoC group. There was no significant difference in the ink propulsion rate of the GRBE-L group compared with the CG group (P > 0.05). It can be seen from the figure that there was a dose relationship in the ink propulsion rates among GRBE-L, GRBE-M and GRBE-H. Through the small intestine motility experiment, it can be known that the fermented liquid of Musa acuminata Colla after fermentation has the effect of promoting the peristalsis of the small intestine of constipated mice.

[0076] (5) Effects of the fermented liquid of Musa acuminata Colla on the liver and kidney coefficients of constipated model mice

[0077] The liver and kidneys were taken, rinsed with cold physiological saline, dried with filter paper, and then the weights of the liver and kidneys of each mouse were weighed using a precision electronic balance and recorded. The liver coefficient and kidney coefficient were calculated according to formula (3) and formula (4) respectively.

[0078]

[0079] The liver coefficients and kidney coefficients of mice in each group after intragastric administration of GRBE for 7 days are shown in Figure 4 shown. As shown in Figure 4It can be seen that there is no significant difference in the liver coefficient and kidney coefficient of each group of mice (P>0.05), which shows that GRBE has no liver and kidney toxicity, and oral administration of GRBE will not cause adverse effects on the liver and kidney of mice, and has a certain degree of safety. (Note: A is the liver coefficient; B is the kidney coefficient)

[0080] (6) Effects of glutinous banana fermentation liquid on serum factor levels in constipation model mice

[0081] After the mouse laxative experiment (A) was completed, blood was collected from the eyeballs of each group of mice and the mice were killed by dislocation of the neck. The serum was naturally precipitated at room temperature. After the serum was precipitated, it was placed in a refrigerated centrifuge at 3000r / min and 4°C for 10 minutes, and the supernatant was taken and frozen in a -80°C refrigerator. The determination of VIP, SP, GAS, MTL and TNF-α in the serum of constipation model mice and the specific steps were carried out according to the instructions of the ELISA kit.

[0082] like Figure 5 As shown in the results, compared with the NG group, the serum VIP content and TNF-α level of mice in the CG group showed an increasing trend (P<0.001). The early intervention of GRBE could reduce the serum VIP, SP and TNF-α levels of mice, and GRBE-H had the best effect (P<0.001). Compared with the NG group, the serum GAS and MTL levels of mice in the CG group were significantly reduced (P<0.001). The early intervention of GRBE at various doses could increase the serum GAS content of mice, among which GRBE-H had the best effect (P<0.001). The changes in the content of various substances in the serum of mice in each group showed that the laxative mechanism of GRBE may be to regulate the secretion of water and electrolytes in the intestine and the peristalsis of the small intestine of mice by down-regulating the VIP and SP content in the serum of mice, up-regulating the serum GAS and MTL content of mice, and down-regulating the serum TNF-α level of mice, thereby improving the intestinal immune dysfunction of mice.

[0083] (7) Effect of glutinous banana fermentation liquid on jejunal pathological changes in constipation model mice

[0084] After the mouse laxative experiment (A) was completed, blood was collected from the eyeballs of each group of mice and the mice were killed by dislocating the neck. The intestinal cavity of the mice was opened to separate the mesentery, and 1 cm of the jejunum of the small intestine was fixed in 4% polyformaldehyde solution (neutral) for more than 2 hours. After dehydration, paraffin embedding, sectioning, hematoxylin-eosin staining and sealing, the pathological changes of the jejunal villi of each group of mice were observed under a microscope and photographed.

[0085] Depend on Figure 6It can be seen that the jejunal villi of the mice in the NG group are arranged compactly, with complete villus morphology and uniform distribution; while for the mice in the CG group, their jejunal villi are shortened, arranged loosely, unevenly distributed, and there is shedding of jejunal villi, indicating that LH can cause constipation in mice and damage the intestinal villi of mice; the jejunal villi of the mice in the MoC group are arranged compactly, with complete villus morphology and uniform distribution, but there is shedding of jejunal villi; the length of the jejunal villi and the arrangement of the villi in the jejunum of the mice in each GRBE group have been improved to varying degrees, and the jejunal villi of the mice in the GRBE-H group are arranged compactly, with longer villus length, complete villus morphology and uniform distribution, and only a few jejunal villi are shed. The conditions of the villi in its jejunum are close to those of the mice in the NG group. It can be seen that as the dose of GRBE increases, its effect of preventing constipation in mice also increases, and GRBE-H has the best effect on preventing constipation in mice.

[0086] In summary, compared with the mice in the CG group, the time to the first black stool excretion of the mice in the GRBE-H and GRBE-M groups is shortened, and the number of black stools excreted within 5 h, the wet weight of the black stools excreted within 5 h, and the water content of the black stools are significantly increased compared with the CG group. The ink propulsion rate of the mice in the GRBE-H and GRBE-M groups is significantly increased compared with the CG group (P < 0.001), and the ink propulsion rate of the mice in the GRBE-H group is close to that of the mice in the MoC group. The results show that GRBE has the effect of promoting small intestine peristalsis, increasing the humidity of feces, helping constipated mice defecate, and relieving the constipation phenomenon of the constipation model mice. From the determination of the content of serum factors, it can be seen that GRBE has the effect of regulating the secretion of water and electrolytes in the intestine of constipated mice and the peristalsis of the small intestine. From the histopathological results of the small intestine villi of the mice in each group, it can be seen that GRBE at low, medium, and high doses all has different degrees of protective effects on the small intestine villi of constipated mice, but GRBE at high dose has a better intestinal protective effect on constipated mice. Thus, it can be seen that the fermented liquid of Musa acuminata has the effects of moistening the intestines and relieving constipation and intestinal protection on the constipation model mice induced by loperamide hydrochloride.

[0087] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention, and they should all be covered within the scope of the present invention.

Claims

1. An application of glutinous banana fermentation liquid, characterized in that: Including, application of the glutinous rice banana fermentation liquid in preparing health food or medicine with bowel-moistening and laxative function; The glutinous banana fermentation liquid has a crude polysaccharide content of 1.34 g / mL and a total acid content of 137.95 g / L, and contains seven organic acids: oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid, maleic acid, and succinic acid.

2. The use according to claim 1, characterized in that: The glutinous banana fermented liquid reduces serum vasoactive intestinal peptide in constipated mice, increases gastrin and motilin, and improves intestinal peristalsis and defecation function.

3. The use according to claim 1, characterized in that: The glutinous banana fermented liquid has the effects of moistening the intestine, promoting bowel movements and protecting the intestines of mice with constipation model induced by loperamide hydrochloride by regulating the secretion of water and electrolytes in the intestines of constipated mice and the peristalsis of the small intestine.

4. A method for preparing glutinous banana fermentation liquid, characterized in that: include, Peeling and pulping the fruit: taking fresh, intact, green-skinned glutinous rice bananas with a fullness of 75% to 80%, peeling and crushing them to obtain glutinous rice banana pulp; Constant temperature enzymolysis: sterilize the glutinous banana pulp at high temperature, add saccharifying enzyme, cellulase, pectinase and lipase after cooling, and perform enzymolysis to obtain glutinous banana pulp enzymolysis liquid; Ingredient fermentation: mix glutinous rice banana pulp enzymatic hydrolyzate, purified water and white sugar, add probiotic solution, control the fermentation temperature at 15-25° C., aerobically ferment for 10-30 days, stir once every 3 days during the fermentation process, until the solid content is 7%-8% and the pH value is about 4, then take the supernatant and filter it to obtain glutinous rice banana fermentation liquid.

5. The preparation method according to claim 4, characterized in that: The isothermal enzymolysis comprises adding 0.05-0.5% saccharifying enzyme, 0.05-0.5% cellulase, 0.05-0.5% pectinase and 0.01-0.05% lipase according to the total weight of the pulp.

6. The preparation method according to claim 4, characterized in that: The glutinous banana pulp enzymatic hydrolysate, purified water and white sugar are mixed, wherein the mass ratio of the glutinous banana pulp enzymatic hydrolysate, purified water and white sugar is 5-10:20-35:2-8 in terms of mass fraction.

7. The preparation method according to claim 4, characterized in that: The probiotic solution is added, wherein the preparation method of the probiotic solution is to use black tea juice as the basic culture solution, add white sugar according to 8-10% of the weight of the black tea juice, and simultaneously add yeast powder, lactobacillus powder, and acetic acid bacteria powder, and culture under aerobic conditions for 10-15 hours.

8. The preparation method according to claim 7, characterized in that: The number of live yeast cells in the probiotic solution is 1.0×10 5 CFU / mL, the number of viable lactobacilli is 1.0×10 6 CFU / mL, the number of viable Acetobacter is 1.0×10 4 CFU / mL.

9. The glutinous banana fermentation liquid obtained by the preparation method according to any one of claims 4 to 8, characterized in that: The glutinous banana fermentation liquid has a crude polysaccharide content of 1.34 g / mL and a total acid content of 137.95 g / L, and contains seven organic acids: oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid, maleic acid, and succinic acid.

10. A beverage with bowel-moistening and laxative function, characterized in that: The beverage uses the glutinous rice and banana fermentation liquid as claimed in claim 9 as an effective ingredient.

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