A prebiotic composition for proliferating intestinal beneficial flora and a method for preparing the same

Abstract

The application discloses a prebiotic composition for proliferating intestinal beneficial bacteria and a preparation method thereof, and comprises the following raw materials in parts by weight: partially hydrolyzed composite glue 35-45 parts, resistant starch of green banana 30-40 parts, enzymatic okra polysaccharide 15-25 parts, and tea polyphenol 3-5 parts. The composition realizes efficient proliferation of intestinal beneficial bacteria and comprehensive and positive regulation of intestinal microenvironment by scientifically compounding partially hydrolyzed composite glue with a specific molecular weight, resistant starch of green banana, enzymatic okra polysaccharide and tea polyphenol. The four components with complementary functions and different effects can not only more efficiently proliferate traditional beneficial bacteria such as bifidobacterium and lactobacillus, but also effectively nourish butyric acid bacteria such as prasrmanella, and inhibit the growth of harmful bacteria, thereby having the functions of significantly improving intestinal flora disorder caused by ampicillin and promoting the generation of short-chain fatty acids and maintaining the intestinal barrier.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of functional food, and particularly relates to a prebiotic composition for proliferating intestinal beneficial flora and a preparation method thereof. BACKGROUND

[0002] The human intestinal tract is the largest and most complex microecosystem in the body, inhabited by trillions of microorganisms, collectively known as the intestinal flora. A healthy intestinal flora can provide essential nutrients for the host through its metabolic activity, build and maintain intestinal barrier function, regulate the host's innate and adaptive immunity, and resist the invasion of pathogenic microorganisms. The balanced and stable structure of the flora is the basis for maintaining human health, otherwise, when affected by factors such as unhealthy eating habits, drug abuse, mental stress or aging, the composition and function of the intestinal flora will be disturbed, showing a decrease in the number of beneficial bacteria (such as Bifidobacterium and Lactobacillus) and an abnormal proliferation of potential pathogenic bacteria (such as Fusobacterium and some Enterobacter). This "ecological imbalance" has been confirmed by a large number of modern medical studies, and is closely related to the occurrence and development of many chronic diseases, including but not limited to functional gastrointestinal diseases such as constipation and diarrhea, inflammatory bowel disease, obesity, metabolic syndrome such as type II diabetes, and even some neurodegenerative diseases and immune system diseases. Therefore, actively intervening and regulating the balance of the intestinal flora through effective means has become a core strategy for maintaining and promoting human health in the fields of modern nutrition and preventive medicine.

[0003] Prebiotics are substances that can selectively stimulate the growth or activation of one or more bacteria in the host's intestinal tract, improve the host's health, and are not digested by the host's gastrointestinal tract. Supplementing prebiotics can promote the proliferation of probiotics in the intestinal tract and improve the structure of the intestinal flora. A single prebiotic does not have a broad spectrum of promoting effects on probiotics, such as xylo-oligosaccharides, which can promote the proliferation of Bifidobacterium adolescentis, Bifidobacterium longum and Lactobacillus salivarius, but have no significant effect on Lactobacillus casei, Lactobacillus fermentum, Bifidobacterium breve, Bifidobacterium infantis and Bifidobacterium bifidum; fructo-oligosaccharides can promote the proliferation of Lactobacillus acidophilus, Lactobacillus salivarius and most Bifidobacterium, but have limited promoting effect on Bifidobacterium bifidum, Lactobacillus casei and Lactobacillus fermentum. Meanwhile, the intestinal microecosystem is a complex community composed of a variety of microorganisms with a large number, and the proliferation of a single flora or a few floras has limited effect on the improvement of the intestinal flora structure. At the same time, part of the prebiotics, especially the oligosaccharides with smaller molecular weight, are rapidly fermented in the proximal colon in an "explosive" manner, which is easy to produce a large amount of gas in a short time, thereby causing discomfort symptoms such as abdominal distension, abdominal pain and excessive gas production.

[0004] Chinese patent application CN107981359A discloses a prebiotic composition and its application. The composition comprises galactooligosaccharides (GOS), fructooligosaccharides (FOS), and lactose isomerization (LOS), wherein the weight ratio of GOS, FOS, and LOS is (5-10):(0.5-1.5):(0.5-1.5), and the degree of polymerization of the FOS ranges from 2 to 9. This prebiotic composition can be used in nutritional foods and pharmaceuticals to enhance the beneficial microorganisms in the human gut microbiota while inhibiting the growth of harmful bacteria, thereby enhancing the body's immunity and overall bodily function. Chinese patent application CN113331335A discloses a prebiotic composition, a solid beverage, and a method for preparing the same. The prebiotic composition comprises the following raw materials in parts by weight: 50-70 parts fructooligosaccharide, 10-30 parts isomaltooligosaccharide, 5-15 parts inulin, 5-10 parts galactooligosaccharide, 2-6 parts stachyose, 2-6 parts xylooligosaccharide, 1-3 parts oat β-glucan, and 1-3 parts chitosan oligosaccharide. This invention, by compounding multiple prebiotics and adjusting the ratios of inulin and isomaltooligosaccharide, as well as the ratios of oat β-glucan and chitosan oligosaccharide, enables synergistic effects among the components, effectively balancing the intestinal flora, significantly increasing beneficial intestinal bacteria, regulating intestinal function, and thus significantly improving constipation. The prebiotic products mentioned above are mostly simple physical mixtures of several known prebiotics (such as inulin and fructooligosaccharides). Although this combination expands the range of bacteria that can be proliferated to some extent, it is essentially a simple superposition of several single mechanisms of action. It does not produce a new structure with synergistic effects at the molecular level, and has limited effect on the proliferation of beneficial bacteria in the gut.

[0005] Therefore, developing a prebiotic composition that can promote the proliferation of diverse core beneficial bacteria and synergistically regulate intestinal immune function is a technical challenge that urgently needs to be solved in this field. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a prebiotic composition for promoting the growth of beneficial intestinal flora and its preparation method.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0009] Partially hydrolyzed composite gum 35-45 parts, green banana resistant starch 30-40 parts, enzymatically hydrolyzed okra polysaccharide 15-25 parts, tea polyphenols 3-5 parts.

[0010] In this invention, the partially hydrolyzed composite gum and resistant starch from green bananas in the prebiotic composition serve as high-quality prebiotics with different fermentation rates. The partially hydrolyzed composite gum acts as a rapid fermentation substrate, primarily utilized by Bifidobacteria and Lactobacillus in the proximal and mid-colon, rapidly improving the gut microbiota structure. The resistant starch from green bananas acts as a slow-release fermentation substrate, successfully transporting energy substances to the distal colon, specifically nourishing anaerobic bacteria such as Clostridium perfringens and Rosella esculenta. The two work synergistically to form a whole-intestinal prebiotic system covering the entire gut from the proximal to the distal colon, achieving broad-spectrum proliferation of core beneficial bacteria such as Bifidobacteria, Lactobacillus, and Clostridium perfringens. Enzymatically hydrolyzed okra polysaccharide, with its unique viscous colloid, physically strengthens the intestinal barrier and directly interacts with intestinal immune cells, exerting immunomodulatory functions. The addition of tea polyphenols further optimizes the microenvironment for gut microbiota colonization through its antioxidant and selective antibacterial effects. Through the synergistic effect of the four ingredients, this composition has a significantly better overall effect than existing single-component or simple physical mixture prebiotic products in restoring gut microbiota diversity, regulating gut immune homeostasis, and maintaining overall gut health.

[0011] Preferably, a prebiotic composition for promoting the growth of beneficial gut microbiota comprises, by weight, the following ingredients:

[0012] Partially hydrolyzed composite gum 35-45 parts, green banana resistant starch 30-40 parts, enzymatically hydrolyzed okra polysaccharide 15-25 parts, tea polyphenols 3-5 parts.

[0013] Preferably, the partially hydrolyzed composite gum is obtained by enzymatic hydrolysis of a mixture of guar gum and flaxseed gum, and its molecular weight is 15,000-25,000 Da.

[0014] In this invention, the inventors discovered that when the molecular weight of the partially hydrolyzed composite gel is between 15,000 and 25,000 Da, it can be continuously fermented throughout the colon (including the proximal and mid-colon) at a more stable rate, thereby enabling the prebiotic composition to have a significant proliferative effect on beneficial bacteria in the gut. When the molecular weight is less than 15,000 Da, the fermentation rate is too fast, and the very short-chain oligosaccharides and monosaccharides are rapidly and violently fermented by microorganisms in the early stages of entering the colon, producing a large amount of gas and causing obvious intestinal intolerance symptoms such as bloating, borborygmus, and increased flatulence. When the molecular weight is higher than 25,000 Da, the excessively long molecular chains and complex spatial structure may hinder the effective contact of glycosidases secreted by bacteria with their internal cleavage sites, resulting in a significant reduction in fermentation efficiency and thus a decrease in the proliferative effect of beneficial bacteria.

[0015] Preferably, the method for preparing the partially hydrolyzed composite adhesive is as follows:

[0016] Guar gum powder and flaxseed gum powder are thoroughly mixed to obtain a composite gum powder. The composite gum powder is then added to deionized water, heated and stirred, followed by the addition of citric acid to adjust the pH. Then, β-mannanase is added for the first enzymatic hydrolysis. After the predetermined hydrolysis time, xylanase, polygalacturonase and pectin lyase are added for the second enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the enzymes are inactivated by boiling water. After cooling to room temperature, the mixture is filtered, and the filtrate is ultrafiltered, concentrated under reduced pressure, and spray-dried to obtain the final product.

[0017] In this invention, guar gum, which has a high viscosity and a simple main chain, is innovatively combined with flaxseed gum, which has a complex structure and is rich in hemicellulose and pectin. This provides a more diverse substrate for enzymatic hydrolysis. The main chain and side chains of flaxseed gum contain a variety of sugar units such as galacturonic acid, rhamnose, arabinose, and xylose. By introducing flaxseed gum, the proliferation of beneficial bacteria can be significantly promoted. Furthermore, under this ratio, the presence of flaxseed gum can moderately regulate the extremely high viscosity of pure guar gum, improve its dispersion and swelling behavior in water, and make the mixture easier to enzymatically hydrolyze, thus playing a role in physical process improvement. Through a specific two-step enzymatic hydrolysis, β-mannanase can cleave the main chain in guar gum and flaxseed gum, thereby reducing the molecular weight. Subsequently, a complex enzyme system, including xylanase, polygalacturonase, and pectin lyase, is added to further degrade the complex side chains and gums in flaxseed gum, resulting in a purer structure and better solubility. The specific enzymatic hydrolysis process of this invention ensures that the molecular weight of the final product remains stable in the range of 15,000-25,000 Da, which is key to achieving slow-release fermentation and avoiding intestinal discomfort.

[0018] Preferably, the mass ratio of guar gum powder to flaxseed gum powder is 7-8:2-3; the heating and stirring temperature is 45-55℃, the time is 40-60 min, and the pH is adjusted to 4-5; the amount of β-mannanase added is 0.3-0.5% of the mass of the composite gum powder, the temperature of the first enzymatic hydrolysis is 45-55℃, and the time is 1-2 h; the amount of xylanase added is 0.1-0.15% of the mass of the composite gum powder; the amount of polygalacturonase added is 0.05-0.07% of the mass of the composite gum powder; the amount of pectin lyase added is 0.03-0.05% of the mass of the composite gum powder; and the temperature of the second enzymatic hydrolysis is 45-55℃, and the time is 2-3 h.

[0019] Preferably, the ultrafiltration process specifically involves: passing the filtrate through a 25000 Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000 Da, and then passing the permeate through a 15000 Da ultrafiltration membrane to obtain a retentate.

[0020] Preferably, the method for preparing the resistant starch from green bananas is as follows:

[0021] Peel the green bananas, slice the pulp, soak it in citric acid solution, then drain, dry, and pulverize to obtain green banana powder. Add the green banana powder to deionized water, quickly blend, then add a compound enzyme, adjust the pH, and perform a first enzymatic hydrolysis. After the enzymatic hydrolysis is complete, filter to obtain green banana starch milk. Heat the green banana starch milk, then add a heat-resistant α-amylase for a second enzymatic hydrolysis. After the enzymatic hydrolysis is complete, cool to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0022] In this invention, a two-step enzymatic hydrolysis process maximizes the protection and high-efficiency purification of the natural granular structure of resistant starch from green bananas under relatively mild conditions. First, enzymatic hydrolysis is performed using a complex enzyme to gently "break down" the starch granules at low temperatures, releasing intact starch granules. Then, under elevated temperatures, a thermoresistant α-amylase is used to degrade the gelatinized non-resistant starch, resulting in high-purity RS2-type resistant starch. Its natural dense semi-crystalline granular structure allows it to effectively resist degradation by gastric acid and small intestinal digestive enzymes, reaching the colon intact. Its fermentation process is gentle and prolonged, enabling it to act as an energy carrier, successfully traversing the proximal and mid-colon, precisely transporting carbon sources to the distal colon, providing efficient and impurity-free carbon sources for bacteria such as Clostridium perfringens, thereby promoting the proliferation of beneficial bacteria in the intestines.

[0023] Preferably, the citric acid solution has a mass concentration of 0.2-0.3%, the soaking temperature is 20-30℃, and the soaking time is 10-15 min; the ratio of green banana powder to deionized water is 1:6-8; the amount of the compound enzyme added is 0.3-0.5% of the mass of the green banana powder; the compound enzyme is composed of cellulase, pectinase, and protease in a mass ratio of 3-4:2-3:1-2; the temperature of the first enzymatic hydrolysis is 40-45℃, the pH is 4.5-5.5, and the time is 2-3 h; the amount of the thermoresistant α-amylase added is 0.1-0.2% of the mass of the green banana powder; the temperature of the second enzymatic hydrolysis is 85-90℃, and the time is 1-1.5 h.

[0024] Preferably, the method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0025] After washing and cutting the okra pods into sections, add them to deionized water and crush them into a pulp. Then add protease and pectinase for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, centrifuge and take the supernatant. Concentrate it under reduced pressure to 1 / 4 of the original volume. Add 4 times the volume of anhydrous ethanol for alcohol precipitation. Wash the precipitate and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0026] In this invention, okra polysaccharides are obtained by enzymatic hydrolysis of okra. After enzymatic modification, the complex side chains of okra polysaccharides are moderately degraded, exposing more bioactive functional groups while retaining the core polysaccharide backbone. When it enters the intestine, it can form a highly biocompatible viscous hydration layer on the surface of the intestinal epithelium. This viscous hydration layer not only reduces the direct impact of pathogens and harmful metabolites on intestinal epithelial cells, but also provides a stable microenvironment for the repair and regeneration of damaged cells, achieving direct and physical reinforcement of the intestinal barrier. Furthermore, as a non-traditional prebiotic substrate, enzymatically hydrolyzed okra polysaccharides can synergistically work with resistant starch and complex gums to help improve the diversity of the gut microbiota structure and jointly build a more diverse and stable gut micro-ecosystem.

[0027] Preferably, the mass ratio of okra pods to deionized water is 1:15-20, the amount of protease added is 0.3-0.4% of the mass of okra pods, the amount of pectinase added is 0.1-0.2% of the mass of okra pods, the enzymatic hydrolysis temperature is 45-55℃, the pH is 5.5-6, and the time is 2-3 hours.

[0028] The present invention also protects a method for preparing a prebiotic composition for increasing beneficial intestinal flora as described above, characterized by comprising the following steps: weighing raw materials according to the formula, mixing them evenly, and thus obtaining the prebiotic composition.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] (1) The prebiotic composition for proliferating beneficial intestinal flora provided by the present invention scientifically combines partially hydrolyzed composite gum of a specific molecular weight, resistant starch of green banana, enzymatically hydrolyzed okra polysaccharide and tea polyphenols to achieve efficient proliferation of beneficial intestinal flora and comprehensive and positive regulation of the intestinal microenvironment. The present invention utilizes four components with complementary functions and different effects to form a whole-chain nutritional system that can cover the proximal colon to the distal colon. It can not only proliferate traditional beneficial bacteria such as Bifidobacterium and Lactobacillus more efficiently, but also effectively nourish butyric acid-producing bacteria such as Clostridium perfringens and inhibit the growth of harmful bacteria. Thus, it has the function of significantly improving the intestinal flora disorder caused by ampicillin, as well as promoting the production of short-chain fatty acids and maintaining the intestinal barrier.

[0031] (2) The prebiotic composition for promoting beneficial gut microbiota provided by the present invention creatively combines guar gum with flaxseed and then performs specific enzymatic hydrolysis to obtain a partially hydrolyzed composite gum that provides gut microbiota with a more comprehensive functional and higher nutritional value prebiotic substrate. This partially hydrolyzed composite gum integrates guar gum rich in β-mannan with flaxseed gum containing both arabinoxylan and rhamnogalacturonic acid through enzymatic hydrolysis. This not only stabilizes the molecular weight of the product in the optimal prebiotic activity range of 15,000-25,000 Da, but more importantly, it has both neutral and acidic oligosaccharide fragments in its molecular structure, forming a diverse range of fermentable substrates. This structural diversity allows it to be utilized by more beneficial strains with different nutritional preferences, resulting in a broader prebiotic effect compared to a single partially hydrolyzed guar gum.

[0032] (3) The prebiotic composition for proliferating beneficial intestinal flora provided by the present invention obtains a green banana resistant starch (RS2 type) with excellent slow-release performance through a specific method. Through two-step enzymatic hydrolysis, the natural granular structure of green banana resistant starch is maximized and highly purified under relatively mild conditions. First, enzymatic hydrolysis is performed by a compound enzyme to gently "break the cell wall" and release complete starch granules at low temperature. Then, under elevated temperature conditions, the gelatinized non-resistant starch is degraded by a thermoresistant α-amylase to obtain high-purity RS2 type resistant starch. Its natural dense semi-crystalline granular structure enables it to effectively resist the degradation of gastric acid and small intestinal digestive enzymes and reach the colon intact. Its fermentation process is slow and long-lasting, which enables it to act as an energy carrier, successfully cross the proximal and middle sections of the colon, and accurately transport carbon source substances to the distal colon, providing efficient and impurity-free carbon source for Clostridium perfringens and other bacteria, thereby proliferating beneficial intestinal flora. Detailed Implementation

[0033] The technical solution of the present invention will be clearly and completely described below 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.

[0034] Unless otherwise specified, all chemical reagents and materials in this invention are purchased from the market or synthesized from raw materials purchased from the market.

[0035] The β-mannanase has an enzyme activity of 10,000 U / g, the xylanase has an enzyme activity of 30,000 U / g, the polygalacturonase has an enzyme activity of 60,000 U / g, the pectin lyase has an enzyme activity of 50,000 U / g, the cellulase, pectinase, and protease have enzyme activities of 60,000 U / g, and the thermostable α-amylase has an enzyme activity of 40,000 U / g.

[0036] Example 1

[0037] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0038] Partially hydrolyzed composite gum (40 parts), green banana resistant starch (35 parts), enzymatically hydrolyzed okra polysaccharide (20 parts), and tea polyphenols (4 parts).

[0039] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0040] 75g of guar gum powder and 25g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 50℃ for 50min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.4g of β-mannanase was added and enzymatically hydrolyzed at 50℃ for 1.5h. After enzymatic hydrolysis, 0.13g of xylanase, 0.06g of polygalacturonase, and 0.04g of pectin lyase were added and enzymatically hydrolyzed at 50℃ for 2.5h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. The permeate was then passed through a 15000Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0041] The preparation method of the resistant starch from green bananas is as follows:

[0042] Peel the green bananas, slice the pulp, and soak it in a 0.25% citric acid solution at 25°C for 13 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 700g of deionized water and quickly blend it. Then, add 0.4g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5), adjust the pH to 5, and enzymatically hydrolyze the mixture at 43°C for 2.5 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.15g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze the mixture at 87°C for 1.5 hours. After enzymatic hydrolysis, cool the mixture to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0043] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0044] Wash and cut 200g of okra pods into sections, add them to 3500g of deionized water, and crush them into a paste. Then add 0.7g of protease and 0.3g of pectinase, and enzymatically hydrolyze at 50℃ and pH 6 for 2.5h. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0045] Example 2

[0046] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0047] Partially hydrolyzed composite gum 35 parts, green banana resistant starch 30 parts, enzymatically hydrolyzed okra polysaccharide 15 parts, tea polyphenols 3 parts.

[0048] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0049] 80g of guar gum powder and 20g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 45℃ for 60min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4. Then, 0.3g of β-mannanase was added and enzymatically hydrolyzed at 45℃ for 2h. After enzymatic hydrolysis, 0.1g of xylanase, 0.05g of polygalacturonase and 0.05g of pectin lyase were added and enzymatically hydrolyzed at 45℃ for 3h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. The permeate was then passed through a 15000Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0050] The preparation method of the resistant starch from green bananas is as follows:

[0051] Peel the green bananas, slice the pulp, and soak it in a 0.2% citric acid solution at 20°C for 15 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 600g of deionized water and quickly blend it. Then, add 0.3g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3:2:1), adjust the pH to 4.5, and enzymatically hydrolyze it at 40°C for 3 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.1g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze it at 85°C for 1.5 hours. After enzymatic hydrolysis, cool the temperature to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0052] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0053] Wash and cut 200g of okra pods into sections, add them to 3000g of deionized water, and crush them into a paste. Then add 0.6g of protease and 0.2g of pectinase, and enzymatically hydrolyze for 3 hours at 45℃ and pH 5.5. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry to obtain enzymatically hydrolyzed okra polysaccharide.

[0054] Example 3

[0055] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0056] Partially hydrolyzed composite gum (45 parts), green banana resistant starch (40 parts), enzymatically hydrolyzed okra polysaccharide (25 parts), and tea polyphenols (5 parts).

[0057] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0058] 70g of guar gum powder and 30g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 55℃ for 40min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 5. Then, 0.5g of β-mannanase was added and enzymatically hydrolyzed at 55℃ for 1h. After enzymatic hydrolysis, 0.15g of xylanase, 0.07g of polygalacturonase and 0.03g of pectin lyase were added and enzymatically hydrolyzed at 55℃ for 2h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. The permeate was then passed through a 15000Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0059] The preparation method of the resistant starch from green bananas is as follows:

[0060] Peel the green bananas, slice the pulp, and soak it in a 0.3% citric acid solution at 30°C for 10 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 800g of deionized water and quickly blend it. Then, add 0.5g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 4:3:2), adjust the pH to 5.5, and enzymatically hydrolyze it at 45°C for 2 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.2g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze it at 90°C for 1 hour. After enzymatic hydrolysis, cool the temperature to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0061] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0062] Wash and cut 200g of okra pods into sections, add them to 4000g of deionized water, and crush them into a paste. Then add 0.8g of protease and 0.4g of pectinase, and enzymatically hydrolyze at 55℃ and pH 6 for 2 hours. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry to obtain enzymatically hydrolyzed okra polysaccharide.

[0063] Comparative Example 1

[0064] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0065] Partially hydrolyzed composite gum (40 parts), green banana resistant starch (35 parts), enzymatically hydrolyzed okra polysaccharide (20 parts), and tea polyphenols (4 parts).

[0066] The preparation method of the aforementioned portion of guar gum is as follows:

[0067] Add 100g of guar gum powder to 2kg of deionized water and heat and stir at 50℃ for 50min. Then add 5wt% citric acid aqueous solution to adjust the pH to 4.5. Next, add 0.4g of β-mannanase and enzymatically hydrolyze at 50℃ for 4h. After enzymatic hydrolysis, boil water to inactivate the enzyme for 10min. After cooling to room temperature, filter. Pass the filtrate through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. Then pass the permeate through a 15000Da ultrafiltration membrane to obtain a retentate. Then concentrate the retentate under reduced pressure at 15kPa and spray dry to obtain the final product.

[0068] The preparation method of the resistant starch from green bananas is as follows:

[0069] Peel the green bananas, slice the pulp, and soak it in a 0.25% citric acid solution at 25°C for 13 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 700g of deionized water and quickly blend it. Then, add 0.4g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5), adjust the pH to 5, and enzymatically hydrolyze the mixture at 43°C for 2.5 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.15g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze the mixture at 87°C for 1.5 hours. After enzymatic hydrolysis, cool the mixture to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0070] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0071] Wash and cut 200g of okra pods into sections, add them to 3500g of deionized water, and crush them into a paste. Then add 0.7g of protease and 0.3g of pectinase, and enzymatically hydrolyze at 50℃ and pH 6 for 2.5h. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0072] Compared with Example 1, no flaxseed gum was added to the partially hydrolyzed composite adhesive in this comparative example.

[0073] Comparative Example 2

[0074] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0075] Partially hydrolyzed composite gum (40 parts), green banana resistant starch (35 parts), enzymatically hydrolyzed okra polysaccharide (20 parts), and tea polyphenols (4 parts).

[0076] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0077] 75g of guar gum powder and 25g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 50℃ for 50min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.4g of β-mannanase was added and enzymatically hydrolyzed at 50℃ for 2.5h. After enzymatic hydrolysis, 0.13g of xylanase, 0.06g of polygalacturonase and 0.04g of pectin lyase were added and enzymatically hydrolyzed at 50℃ for 3.5h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was ultrafiltered through a 15000Da membrane to obtain a permeate with a molecular weight less than 15000Da. The permeate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0078] The preparation method of the resistant starch from green bananas is as follows:

[0079] Peel the green bananas, slice the pulp, and soak it in a 0.25% citric acid solution at 25°C for 13 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 700g of deionized water and quickly blend it. Then, add 0.4g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5), adjust the pH to 5, and enzymatically hydrolyze the mixture at 43°C for 2.5 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.15g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze the mixture at 87°C for 1.5 hours. After enzymatic hydrolysis, cool the mixture to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0080] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0081] Wash and cut 200g of okra pods into sections, add them to 3500g of deionized water, and crush them into a paste. Then add 0.7g of protease and 0.3g of pectinase, and enzymatically hydrolyze at 50℃ and pH 6 for 2.5h. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0082] Compared with Example 1, the molecular weight of the partially hydrolyzed composite adhesive prepared in this comparative example is less than 15000 Da.

[0083] Comparative Example 3

[0084] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0085] Partially hydrolyzed composite gum (40 parts), green banana resistant starch (35 parts), enzymatically hydrolyzed okra polysaccharide (20 parts), and tea polyphenols (4 parts).

[0086] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0087] 75g of guar gum powder and 25g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 50℃ for 50min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.4g of β-mannanase was added and enzymatically hydrolyzed at 50℃ for 0.5h. After enzymatic hydrolysis, 0.13g of xylanase, 0.06g of polygalacturonase and 0.04g of pectin lyase were added and enzymatically hydrolyzed at 50℃ for 1.5h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a retentate with a molecular weight greater than 25000Da. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0088] The preparation method of the resistant starch from green bananas is as follows:

[0089] Peel the green bananas, slice the pulp, and soak it in a 0.25% citric acid solution at 25°C for 13 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 700g of deionized water and quickly blend it. Then, add 0.4g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5), adjust the pH to 5, and enzymatically hydrolyze the mixture at 43°C for 2.5 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.15g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze the mixture at 87°C for 1.5 hours. After enzymatic hydrolysis, cool the mixture to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0090] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0091] Wash and cut 200g of okra pods into sections, add them to 3500g of deionized water, and crush them into a paste. Then add 0.7g of protease and 0.3g of pectinase, and enzymatically hydrolyze at 50℃ and pH 6 for 2.5h. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0092] Compared with Example 1, the partially hydrolyzed composite adhesive prepared in this comparative example has a molecular weight greater than 25,000 Da.

[0093] Comparative Example 4

[0094] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0095] 40 parts of partially hydrolyzed composite gum, 20 parts of enzymatically hydrolyzed okra polysaccharide, and 4 parts of tea polyphenols.

[0096] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0097] 75g of guar gum powder and 25g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 50℃ for 50min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.5g of β-mannanase was added and enzymatically hydrolyzed at 50℃ for 1.5h. After enzymatic hydrolysis, 0.13g of xylanase, 0.06g of polygalacturonase and 0.04g of pectin lyase were added and enzymatically hydrolyzed at 50℃ for 2.5h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. The permeate was then passed through a 15000Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0098] The method for preparing the enzymatically hydrolyzed okra polysaccharide is as follows:

[0099] Wash and cut 200g of okra pods into sections, add them to 3500g of deionized water, and crush them into a paste. Then add 0.7g of protease and 0.3g of pectinase, and enzymatically hydrolyze at 50℃ and pH 6 for 2.5h. After enzymatic hydrolysis, centrifuge, take the supernatant, concentrate it under reduced pressure to 1 / 4 of the original volume, add 4 times the volume of anhydrous ethanol for alcohol precipitation, wash the precipitate, and vacuum dry it to obtain enzymatically hydrolyzed okra polysaccharide.

[0100] Compared to Example 1, the prebiotic composition in this comparative example did not contain green banana resistant starch.

[0101] Comparative Example 5

[0102] A prebiotic composition for promoting the growth of beneficial gut microbiota, comprising, by weight, the following ingredients:

[0103] Partially hydrolyzed composite gum (40 parts), green banana resistant starch (35 parts), and tea polyphenols (4 parts).

[0104] The preparation method of the partially hydrolyzed composite adhesive is as follows:

[0105] 75g of guar gum powder and 25g of flaxseed gum powder were thoroughly mixed to obtain a composite gum powder. The composite gum powder was then added to 2kg of deionized water and heated and stirred at 50℃ for 50min. Subsequently, 5wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.5g of β-mannanase was added and enzymatically hydrolyzed at 50℃ for 1.5h. After enzymatic hydrolysis, 0.13g of xylanase, 0.06g of polygalacturonase and 0.04g of pectin lyase were added and enzymatically hydrolyzed at 50℃ for 2.5h. After enzymatic hydrolysis, the enzymes were inactivated by boiling water for 10min. After cooling to room temperature, the mixture was filtered. The filtrate was passed through a 25000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25000Da. The permeate was then passed through a 15000Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15kPa and spray-dried to obtain the final product.

[0106] The preparation method of the resistant starch from green bananas is as follows:

[0107] Peel the green bananas, slice the pulp, and soak it in a 0.25% citric acid solution at 25°C for 13 minutes. Then, remove the bananas, drain the water, dry them, and pulverize them to obtain green banana powder. Add 100g of green banana powder to 700g of deionized water and quickly blend it. Then, add 0.4g of a compound enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5), adjust the pH to 5, and enzymatically hydrolyze the mixture at 43°C for 2.5 hours. After enzymatic hydrolysis, filter the mixture to obtain green banana starch milk. Heat the green banana starch milk to 85°C, then add 0.15g of thermoresistant α-amylase, adjust the pH to 6, and enzymatically hydrolyze the mixture at 87°C for 1.5 hours. After enzymatic hydrolysis, cool the mixture to below 60°C, centrifuge at high speed, collect the precipitate, wash and dry the precipitate to obtain the final product.

[0108] Compared to Example 1, the prebiotic composition in this comparative example did not contain enzymatically hydrolyzed okra polysaccharide.

[0109] The prebiotic compositions prepared in Examples 1-3 and Comparative Examples 1-5 were subjected to mouse experiments, as detailed below:

[0110] One hundred SPF-grade male BALB / c mice were randomly divided into 10 groups of 10 mice each. The BALB / c mice, 6-8 weeks old and weighing 18-22g, were purchased from Chongqing Tengxin Biotechnology Co., Ltd. The experimental animals were housed in a barrier environment animal room at 22℃ and 40-70% humidity, with 12-hour light-dark cycles. They were fed standard feed for one week as part of an acclimatization program and had free access to water. The blank control group was administered an equal volume of physiological saline by gavage. The other groups were first treated with ampicillin antibiotics to disrupt the gut microbiota, followed by continuous gavage administration of 300 μL ampicillin (1 mg / mL) once daily for 14 days. After the model was established, the prebiotic composition for each group was prepared into a 25 mg / mL solution. Groups in Examples 1-3 were then administered the prebiotic composition from Examples 1-3, and groups in Comparative Examples 1-5 were administered the prebiotic composition from Comparative Examples 1-5. The gavage dose for each group was 0.4 mL (500 mg / kg bw). The blank control group and the model group were administered an equal volume of physiological saline by gavage. The test samples were administered for 30 days. The initial fasting body weight and the fasting body weight at the end of the experiment were recorded, and the results are shown in Table 1 below.

[0111] Table 1. Changes in mouse body weight

[0112]

[0113] As shown in Table 1 above, compared with the blank group, the weight gain in the model group was significantly reduced, indicating that the model group was successfully established. The weight gain of mice in Examples 1-3 was significantly higher than that in the model group, indicating that the prebiotic composition prepared in this invention can significantly promote the recovery and growth of mouse weight. At the same time, Examples 1-3 were superior to Comparative Examples 1-5, indicating that the raw materials of the prebiotic composition of this invention have a synergistic effect.

[0114] Twenty-four hours after gavage administration of the prebiotic composition on days 0 and 30, respectively, 0.1 g of fecal matter was aseptically collected from the anus of mice in the control group, model group, Example 1 group, and Comparative Examples 1-5 groups. The fecal matter was placed in a dry, sterile test tube and diluted to 10 μL with sterile diluent. -6 The bacteria were inoculated onto culture media containing Bifidobacterium, Lactobacillus, Enterobacter, Enterococcus, Clostridium perfringens, and Clostridium praosporum, respectively. The culture medium for Bifidobacterium was BBL agar, for Lactobacillus it was LBS agar, for Enterobacterium it was eosin methylene blue agar, for Enterococcus it was sodium azide-crystal violet-esculin agar, for Clostridium perfringens it was TSC agar, and for Clostridium praosporum it was YCFA liquid medium. The cultures were incubated under anaerobic conditions for 48 hours. The number of colonies per gram of wet stool was counted and calculated, and the logarithm was taken. The results are shown in Table 2 below.

[0115] Table 2. Effects of each group of prebiotic compositions on the gut microbiota of mice ( ±s, log cfu / g, n=10)

[0116]

[0117]

[0118] Note: # p<0.05, compared with the control group, ## p<0.01, compared with the blank group; *p<0.05, compared with the model group; **p<0.01, compared with the model group.

[0119] As shown in Table 2 above, compared with the blank group, the feces of mice in the model group showed a significant increase in Enterococcus, Enterobacter, and Clostridium perfringens, while Lactobacillus, Bifidobacterium, and Clostridium plasminogen activator were significantly reduced. This indicates that ampicillin altered the composition of the intestinal flora in mice, resulting in successful biofilm formation. Compared with the model group, the fecal flora of mice fed in Example 1 showed a significant increase in Bifidobacterium, Lactobacillus, and Clostridium plasminogen activator, while Clostridium perfringens was significantly reduced. This indicates that the prebiotic composition prepared in this invention increases the content of beneficial bacteria and restores Enterococcus and Enterobacter to normal levels, demonstrating a significant restorative effect on intestinal flora imbalance and maintaining the balance of the intestinal microbiota. Furthermore, compared with Comparative Examples 1-5, Example 1 showed a significant increase in the content of Bifidobacterium, Lactobacillus, and Clostridium plasminogen activator, indicating a synergistic effect among the raw materials in the prebiotic composition of this invention.

[0120] The prebiotic compositions prepared in Example 1 and Comparative Examples 1-5 were subjected to in vitro fermentation experiments, and the changes in their gas production were measured, as follows:

[0121] Preparation of basic anaerobic culture medium: yeast extract (2.0 g), peptone (2.0 g), NaCl (0.1 g), K2HPO4 (0.04 g), KH2PO4 (0.01 g), CaCl2·2H2O (0.01 g), MgSO4·7H2O (0.01 g), NaHCO3 (2.0 g), heme (0.02 g), L-cysteine ​​(0.5 g), bile salts (0.5 g), Tween 80 (2.0 mL), resazurite solution (1.0 mL, 1%, w / v), vitamin K (10.0 μL) and ultrapure water. Adjust the pH of the basic nutrient medium to pH = 7.0 and autoclave at 121℃ for 15 min.

[0122] Phosphate-buffered saline (PBS) preparation: Dissolve sodium chloride (2.0 g), potassium dihydrogen phosphate (0.2 g), disodium hydrogen phosphate (2.9 g), and potassium chloride (0.2 g) in 1 L of ultrapure water, sterilize, cool, adjust pH to 7.4, and store at 4 °C for later use.

[0123] On the day of the experiment, feces were collected from healthy adults (one male and one female) (who were in good health, had no gastrointestinal diseases, and had not taken antibiotics in the past three months). The collected fecal samples were transferred to an anaerobic incubator for processing. 10g of each fecal sample was weighed on an electronic balance and mixed with phosphate buffer solution at a ratio of 1:10 (w / v) (using one male and one female sample to obtain a richer bacterial community in order to construct an intestinal flora model). The mixture was then filtered through gauze for later use.

[0124] Six Erlenmeyer flasks were prepared, and 27 mL of anaerobic culture medium, 3 mL of filtered fecal suspension, and 1 g of the prebiotic composition from Example 1 and Comparative Examples 1-5 were added to each flask. The blank control group used an equal volume of sterile water instead of the prebiotic composition. The flasks were incubated in a 37°C anaerobic incubator. Gas production was measured using a graduated syringe at 4, 8, 12, 16, 20, and 24 h of fermentation. The results are shown in Table 3 below.

[0125] Table 3. Effect of different prebiotic compositions on gas production (mL)

[0126]

[0127] As can be seen from Table 3 above, the prebiotic composition prepared by the present invention (Example 1) has a moderate gas production rate in the early stage (4-12h) and a stable gas production rate in the later stage (12-24h), which can slow down the gas production rate of fermentation in the intestine and reduce the discomfort caused by bloating. However, due to the decrease in molecular weight of the partially hydrolyzed composite gum in Comparative Example 2 and the absence of resistant starch from green bananas in Comparative Example 4, the gas production rate in the early stage is significantly increased, producing a large amount of gas, which can easily lead to obvious intestinal intolerance symptoms such as bloating, borborygmus, and increased flatulence.

[0128] The above description is a further detailed explanation of the present invention in conjunction with specific implementation examples. It should not be considered that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, several simple deductions or substitutions can be made without departing from the concept of the present invention, and all such deductions or substitutions should be considered to fall within the protection scope of the present invention.

[0129] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A prebiotic composition for promoting the growth of beneficial intestinal flora, characterized in that, By weight, it includes the following ingredients: Partially hydrolyzed composite gum 35-45 parts, green banana resistant starch 30-40 parts, enzymatically hydrolyzed okra polysaccharide 15-25 parts, tea polyphenols 3-5 parts; The partially hydrolyzed composite gum is obtained by enzymatic hydrolysis of guar gum and flaxseed gum, with a molecular weight of 15000-25000 Da. The preparation method of the partially hydrolyzed composite gum is as follows: Guar gum powder and flaxseed gum powder are thoroughly mixed to obtain composite gum powder. Then, the composite gum powder is added to deionized water, heated and stirred, followed by the addition of citric acid to adjust the pH, and then β-mannanase is added for a first enzymatic hydrolysis. After the enzymatic hydrolysis reaches a predetermined time, xylanase, polygalacturonase and pectin lyase are added for a second enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the enzymes are inactivated by boiling water, cooled to room temperature and filtered. The filtrate is then subjected to ultrafiltration, concentrated under reduced pressure, and spray-dried to obtain the final product. The preparation method of the resistant starch from green bananas is as follows: Peel the green bananas, slice the pulp, soak them in citric acid solution, then remove them, drain the water, dry them, and pulverize them to obtain green banana powder; add the green banana powder to deionized water, quickly blend it, then add a compound enzyme, adjust the pH, and perform a first enzymatic hydrolysis. After the enzymatic hydrolysis is completed, filter to obtain green banana starch milk; heat the green banana starch milk, then add a thermoresistant α-amylase for a second enzymatic hydrolysis. After the enzymatic hydrolysis is completed, cool it to below 60°C, centrifuge it at high speed, collect the precipitate, wash and dry the precipitate to obtain the starch; the compound enzyme is composed of cellulase, pectinase, and protease in a mass ratio of 3-4:2-3:1-2. The preparation method of the enzymatic hydrolyzed okra polysaccharide is as follows: okra pods are washed, cut into sections, and added to deionized water. They are then crushed into a pulp, followed by the addition of protease and pectinase for enzymatic hydrolysis. After enzymatic hydrolysis, the pods are centrifuged, and the supernatant is collected and concentrated under reduced pressure to 1 / 4 of the original volume. Four times the volume of anhydrous ethanol is added for alcohol precipitation. The precipitate is washed and dried under vacuum to obtain the enzymatic hydrolyzed okra polysaccharide.

2. The prebiotic composition for promoting beneficial intestinal flora according to claim 1, characterized in that, By weight, it includes the following ingredients: Partially hydrolyzed composite gum 40-45 parts, green banana resistant starch 30-35 parts, enzymatically hydrolyzed okra polysaccharide 20-25 parts, tea polyphenols 4-5 parts.

3. The prebiotic composition for promoting beneficial intestinal flora according to claim 1, characterized in that, In the preparation of the partially hydrolyzed composite gum, the mass ratio of guar gum powder to flaxseed gum powder is 7-8:2-3; the heating and stirring temperature is 45-55℃, the time is 40-60 min, and the pH is adjusted to 4-5; the amount of β-mannanase added is 0.3-0.5% of the mass of the composite gum powder, the temperature of the first enzymatic hydrolysis is 45-55℃, and the time is 1-2 h; the amount of xylanase added is 0.1-0.15% of the mass of the composite gum powder; the amount of polygalacturonase added is 0.05-0.07% of the mass of the composite gum powder; the amount of pectin lyase added is 0.03-0.05% of the mass of the composite gum powder; the temperature of the second enzymatic hydrolysis is 45-55℃, and the time is 2-3 h.

4. The prebiotic composition for promoting beneficial intestinal flora according to claim 1, characterized in that, In the preparation process of the resistant starch from green bananas, the mass concentration of the citric acid solution is 0.2-0.3%, the soaking temperature is 20-30℃, and the time is 10-15 min; the material-to-liquid ratio of green banana powder to deionized water is 1:6-8; the amount of the compound enzyme added is 0.3-0.5% of the mass of the green banana powder; the temperature of the first enzymatic hydrolysis is 40-45℃, the pH is 4.5-5.5, and the time is 2-3 h; the amount of the thermoresistant α-amylase added is 0.1-0.2% of the mass of the green banana powder; the temperature of the second enzymatic hydrolysis is 85-90℃, and the time is 1-1.5 h.

5. The prebiotic composition for promoting beneficial intestinal flora according to claim 1, characterized in that, In the preparation process of the enzymatic hydrolysis of okra polysaccharide, the mass ratio of okra pods to deionized water is 1:15-20, the amount of protease added is 0.3-0.4% of the mass of okra pods, the amount of pectinase added is 0.1-0.2% of the mass of okra pods, the enzymatic hydrolysis temperature is 45-55℃, the pH is 5.5-6, and the time is 2-3 hours.

6. A method for preparing a prebiotic composition for proliferating beneficial intestinal flora as described in any one of claims 1-5, characterized in that, Includes the following steps: Weigh the ingredients according to the formula, mix them evenly, and you have the product.

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