Prebiotics composition capable of proliferating intestinal beneficial microbial communities and preparation method of prebiotics composition

Abstract

The invention discloses a prebiotic composition capable of proliferating intestinal beneficial flora and a preparation method thereof, and the prebiotic composition comprises the following raw materials in parts by weight: 35-45 parts of partially hydrolyzed complex gum, 30-40 parts of green banana resistant starch, 15-25 parts of enzymolysis okra polysaccharide and 3-5 parts of tea polyphenol. According to the composition, the partially hydrolyzed complex gum with specific molecular weight, the green banana resistant starch, the enzymolysis okra polysaccharide and the tea polyphenol are scientifically compounded, so that efficient proliferation of intestinal effective microbial communities and comprehensive and positive regulation and control of intestinal microenvironment are realized, and the composition is prepared by utilizing four components with complementary functions and different effects. The bacillus amyloliquefaciens can effectively proliferate traditional beneficial bacteria such as bifidobacteria and lactobacillus, can effectively nourish butyric acid producing bacteria such as clostridium praeparatum and inhibit growth of harmful bacteria, so that the bacillus amyloliquefaciens has the function of remarkably improving intestinal flora disorder caused by ampicillin, and has the functions of promoting generation of short-chain fatty acid and maintaining intestinal barriers.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional foods, and in particular relates to a prebiotic composition for proliferating beneficial intestinal flora and a preparation method thereof. Background Art

[0002] The human intestine is the body's largest and most complex microbial ecosystem, home to trillions of microorganisms, collectively known as the intestinal microbiota. Through their metabolic activities, a healthy intestinal microbiota provides essential nutrients to the host, builds and maintains the intestinal barrier function, regulates the host's innate and adaptive immunity, and protects against pathogenic microorganisms. A balanced and stable microbial structure is fundamental to maintaining human health. Conversely, when compromised by factors such as poor diet, substance abuse, stress, or aging, the composition and function of the intestinal microbiota can become disrupted, manifesting as a decrease in the number of beneficial bacteria (such as Bifidobacteria and Lactobacilli) and an abnormal proliferation of potentially pathogenic bacteria (such as Fusobacteria and some Enterobacteriaceae). This "dysbiosis" has been confirmed by numerous modern medical studies and is closely associated with the development and progression of numerous chronic diseases, including but not limited to functional gastrointestinal disorders such as constipation and diarrhea, inflammatory bowel disease, obesity, metabolic syndromes such as type 2 diabetes, and even some neurodegenerative and immune disorders. Therefore, actively intervening and regulating the balance of the intestinal microbiota through effective means has become a core strategy in modern nutrition and preventive medicine for maintaining and promoting human health.

[0003] Prebiotics are substances that selectively stimulate the growth or activation of one or more bacteria in the host's intestines, improving host health without being digested by the host's gastrointestinal tract. Supplementation with prebiotics can promote the proliferation of probiotics in the intestines and improve the structure of the intestinal flora. The promoting effect of a single prebiotic on probiotics is not broad-spectrum. For example, oligoxylose can promote the proliferation of Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus salivarius, but has no significant effect on Lactobacillus casei, Lactobacillus fermentum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium bifidum; oligofructose can promote the proliferation of Lactobacillus acidophilus, Lactobacillus salivarius, and most bifidobacteria, but has limited effect on Bifidobacterium bifidum, Lactobacillus casei, and Lactobacillus fermentum. The intestinal microbiome is a complex community composed of a large number of microorganisms of various types. The proliferation of a single bacterial group or a small number of bacterial groups has a relatively limited effect on improving the structure of the intestinal flora. At the same time, some prebiotics, especially oligosaccharides with smaller molecular weight, are fermented "explosively" rapidly in the proximal colon, which can easily produce a large amount of gas in a short period of time, thereby causing uncomfortable 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 galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), and isomerized lactose (LOS), with the weight ratio of galacto-oligosaccharides, fructo-oligosaccharides, and isomerized lactose (LOS) being (5-10): (0.5-1.5): (0.5-1.5), wherein the degree of polymerization of the fructo-oligosaccharides ranges from 2 to 9. The prebiotic composition can be used in nutritional foods and pharmaceuticals to enhance the health-promoting microorganisms in the human intestinal flora while inhibiting the growth of harmful bacteria, thereby enhancing the body's immunity and overall function. Chinese patent application CN113331335A discloses a prebiotic composition, a solid beverage, and a preparation method thereof. The prebiotic composition comprises the following raw materials in parts by weight: 50-70 parts of oligofructose, 10-30 parts of oligomaltodextrose, 5-15 parts of inulin, 5-10 parts of oligogalactose, 2-6 parts of stachyose, 2-6 parts of oligoxylose, 1-3 parts of oat beta-glucan, and 1-3 parts of oligochitosan. This invention combines multiple prebiotics and adjusts the ratio of prebiotics inulin and oligomaltodextrose, as well as the ratio of oat beta-glucan and oligochitosan, to achieve synergistic effects between the components, effectively balance the intestinal flora, significantly proliferate beneficial intestinal bacteria, regulate intestinal function, and thus significantly improve constipation. The prebiotic products of the above patents are mostly simple physical mixtures of several known prebiotics (such as inulin + oligofructose). Although this combination expands the range of proliferated bacteria to a certain extent, it is essentially still 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 intestines.

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

[0006] In view of the shortcomings of the prior art, the object of the present invention is to provide a prebiotic composition for proliferating beneficial intestinal bacteria and a preparation method thereof.

[0007] To achieve the above object, the present invention provides the following technical solutions: A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 35-45 parts of partially hydrolyzed composite gum, 30-40 parts of green banana resistant starch, 15-25 parts of enzymatically hydrolyzed okra polysaccharide, and 3-5 parts of tea polyphenols.

[0008] In the present invention, the partially hydrolyzed composite gum and green banana resistant starch in the prebiotic composition serve as high-quality prebiotics with different fermentation rates. The partially hydrolyzed composite gum serves as a fast fermentation substrate, which is mainly rapidly utilized by bifidobacteria, lactobacilli, etc. in the proximal and middle parts of the colon, thereby rapidly improving the structure of the flora. The green banana resistant starch serves as a slow-release fermentation substrate, which successfully transports energy substances to the distal end of the colon, specifically nourishing anaerobic bacteria such as Faecalibacterium prausnitzii and Roseburia. The two act synergistically to form a whole-intestinal prebiotic system that can cover from the proximal to the distal end of the colon, thereby achieving a broad-spectrum proliferation of core beneficial flora such as bifidobacteria, lactobacilli and Faecalibacterium prausnitzii. Enzymatically hydrolyzed okra polysaccharide, with its unique viscous colloid, physically strengthens the intestinal barrier, and directly interacts with intestinal immune cells to exert an immune regulatory function. The addition of tea polyphenols further optimizes the microenvironment for flora colonization through its antioxidant and selective antibacterial effects. Through the synergistic effect of the four raw materials, the comprehensive effect of this composition in restoring intestinal flora diversity, regulating intestinal immune homeostasis and maintaining overall intestinal health is significantly better than existing single-component or simple physically mixed prebiotic products.

[0009] Preferably, a prebiotic composition for proliferating beneficial intestinal bacteria comprises the following raw materials in parts by weight: 35-45 parts of partially hydrolyzed composite gum, 30-40 parts of green banana resistant starch, 15-25 parts of enzymatically hydrolyzed okra polysaccharide, and 3-5 parts of tea polyphenols.

[0010] Preferably, the partially hydrolyzed composite gum is obtained by mixing guar gum and flaxseed gum and then subjecting them to enzymatic hydrolysis, and has a molecular weight of 15,000-25,000 Da.

[0011] In the present invention, the inventors found that when the molecular weight of the partially hydrolyzed composite gum is between 15,000 and 25,000 Da, it can be continuously fermented at a more stable rate throughout the entire colon (including the proximal and middle sections), thereby enabling the prebiotic composition to have a significant proliferative effect on the beneficial bacteria in the intestine; when the molecular weight is less than 15,000 Da, its fermentation rate is too fast, and the very short-chain oligosaccharides and monosaccharides will be rapidly and vigorously fermented by microorganisms in the early stage of entering the colon, producing a large amount of gas, leading to obvious intestinal intolerance symptoms such as abdominal distension, intestinal rumbling, and increased flatulence; when the molecular weight is higher than 25,000 Da, the excessively long molecular chain and complex spatial structure may hinder the glycosidase secreted by the bacteria from effectively contacting the enzymatic cleavage sites inside it, resulting in a significant reduction in fermentation efficiency, thereby reducing the proliferation effect of beneficial bacteria.

[0012] Preferably, the preparation method of the partially hydrolyzed composite glue is as follows: Guar gum powder and flaxseed gum powder are fully mixed to obtain composite gum powder, and then the composite gum powder is added to deionized water, heated and stirred, and then citric acid is added to adjust the pH, and then β-mannanase is added to perform a primary enzymolysis. After the enzymolysis reaches a predetermined time, xylanase, polygalacturonase and pectin lyase are added to perform a secondary enzymolysis. After the enzymolysis is completed, the enzyme is inactivated with boiling water, and the mixture is filtered after cooling to room temperature. The filtrate is ultrafiltered, concentrated under reduced pressure, and spray-dried to obtain the product.

[0013] In the present invention, guar gum with high viscosity and a single main chain is innovatively compounded with flaxseed gum with a complex structure and rich in hemicellulose and pectin, providing a substrate with more diverse functions for enzymatic hydrolysis. The main chain and side chains of flaxseed gum contain multiple sugar units such as galacturonic acid, rhamnose, arabinose, and xylose. The introduction of flaxseed gum can significantly promote the proliferation of beneficial bacteria. Moreover, 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, make the mixed material easier to enzymatic hydrolysis, and play a physical process improvement role. Through a specific two-step enzymatic hydrolysis, β-mannanase can cut the main chains in guar gum and flaxseed gum, reducing the main molecular weight. Subsequently, a complex enzyme such as xylanase, polygalacturonase and pectin lyase is added to further degrade the complex side chains and gum in the flaxseed gum, making the structure purer and the solubility better. The specific enzymatic hydrolysis process of the present invention ensures that the molecular weight of the final product is stable in the range of 15,000-25,000 Da, which is the key to achieving slow-release fermentation and avoiding intestinal discomfort.

[0014] Preferably, the mass ratio of the guar gum powder to the flaxseed gum powder is 7-8:2-3, the heating and stirring temperature is 45-55°C, the time is 40-60 min, and the pH is adjusted to 4-5; the amount of the β-mannanase added is 0.3-0.5% of the mass of the composite rubber powder, the temperature of the primary enzymatic hydrolysis is 45-55°C, and the time is 1-2 h; the amount of the xylanase added is 0.1-0.15% of the mass of the composite rubber powder, the amount of the polygalacturonase added is 0.05-0.07% of the mass of the composite rubber powder, the amount of the pectin lyase added is 0.03-0.05% of the mass of the composite rubber powder, the temperature of the secondary enzymatic hydrolysis is 45-55°C, and the time is 2-3 h.

[0015] Preferably, the ultrafiltration treatment is specifically as follows: passing the filtrate through a 25,000Da ultrafiltration membrane to obtain a permeate with a molecular weight less than 25,000Da, and then passing the permeate through a 15,000Da ultrafiltration membrane to obtain a retentate.

[0016] Preferably, the preparation method of the green banana resistant starch is as follows: The green bananas are peeled, the pulp is sliced, and then immersed in a citric acid solution, then removed, drained, dried, and crushed to obtain green banana powder; the green banana powder is added to deionized water, quickly pulped, and then a complex enzyme is added to adjust the pH to perform a primary enzymatic hydrolysis, and filtered after the enzymatic hydrolysis is completed to obtain green banana starch milk; the green banana starch milk is heated, and then a high-temperature resistant α-amylase is added to perform a secondary enzymatic hydrolysis, and after the enzymatic hydrolysis is completed, the temperature is cooled to below 60°C, and high-speed centrifugation is performed to collect the precipitate, and the precipitate is washed and dried to obtain the product.

[0017] In the present invention, through two-step enzymatic hydrolysis, the maximum protection and high-efficiency purification of the natural granular structure of green banana resistant starch are achieved under relatively mild conditions. First, enzymatic hydrolysis is carried out by a complex enzyme to gently "break the wall" and release complete starch granules at low temperature. Then, under elevated temperature conditions, high-temperature resistant α-amylase is used to degrade the gelatinized non-resistant starch to obtain high-purity RS2 resistant starch. Its natural dense semi-crystalline granular structure enables it to effectively resist degradation by gastric acid and small intestinal digestive enzymes and reach the colon intact. Its fermentation process is smooth and long-lasting, allowing it to serve as an energy carrier and successfully cross the proximal and middle parts of the colon, accurately transporting carbon source substances to the distal end of the colon, providing an efficient and impurity-free carbon source for Faecalibacterium prausnitzii and the like, thereby proliferating the beneficial bacteria in the intestine.

[0018] Preferably, the mass concentration of the citric acid solution is 0.2-0.3%, the immersion temperature is 20-30°C, and the immersion time is 10-15 min; the material-liquid ratio of the green banana powder to deionized water is 1:6-8, the added amount of the complex enzyme is 0.3-0.5% of the mass of the green banana powder, the complex enzyme is composed of cellulase, pectinase, and protease in a mass ratio of 3-4:2-3:1-2, the temperature of the primary enzymatic hydrolysis is 40-45°C, the pH is 4.5-5.5, and the time is 2-3 h; the added amount of the high-temperature resistant α-amylase is 0.1-0.2% of the mass of the green banana powder, the temperature of the secondary enzymatic hydrolysis is 85-90°C, and the time is 1-1.5 h.

[0019] Preferably, the preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: The okra pods are cleaned and cut into segments, added into deionized water, and mashed into a pulp. Subsequently, protease and pectinase are added for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the mixture is centrifuged, the supernatant is taken, 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 vacuum dried to obtain enzymatically hydrolyzed okra polysaccharides.

[0020] In the present invention, okra is subjected to a composite enzymatic hydrolysis to obtain okra polysaccharide. After enzymatic modification, the complex side chains of okra polysaccharide are moderately degraded, while retaining its core polysaccharide skeleton, more biologically active functional groups are exposed. When it enters the intestine, it can form a viscous hydrated layer with high biocompatibility on the surface of the intestinal epithelium. This viscous hydrated layer can not only reduce the direct impact of pathogens and harmful metabolites on intestinal epithelial cells, but also provide a stable microenvironment for the repair and regeneration of damaged cells, thereby achieving direct and physical reinforcement of the intestinal barrier. Moreover, enzymatically hydrolyzed okra polysaccharide, as a non-traditional prebiotic substrate, can synergize with resistant starch and composite gum, help to enhance the diversity of the bacterial flora structure, and jointly construct a more diverse and more stable intestinal microecological system.

[0021] Preferably, the mass ratio of the okra pods to deionized water is 1:15-20, the amount of the protease added is 0.3-0.4% of the mass of the okra pods, the amount of the pectinase added is 0.1-0.2% of the mass of the okra pods, the temperature of the enzymolysis is 45-55°C, the pH is 5.5-6, and the time is 2-3h.

[0022] The present invention also provides a method for preparing the prebiotic composition for proliferating beneficial intestinal bacteria, which is characterized by comprising the following steps: weighing raw materials according to a formula, and mixing them uniformly to obtain the prebiotic composition.

[0023] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention provides a prebiotic composition for proliferating beneficial intestinal flora. The composition is scientifically compounded with partially hydrolyzed composite gum of a specific molecular weight, green banana resistant starch, enzymatically hydrolyzed okra polysaccharide and tea polyphenols, thereby achieving efficient proliferation of beneficial intestinal flora and comprehensive, positive regulation of the intestinal microenvironment. The present invention utilizes four components with complementary functions and different effects to form a full-chain nutritional system that can cover the entire colon from the proximal colon to the distal colon. It can not only more efficiently proliferate traditional beneficial bacteria such as Bifidobacterium and Lactobacillus, but also effectively nourish butyrate-producing bacteria such as Faecalibacterium prausnitzii and inhibit the growth of harmful bacteria, thereby significantly improving the intestinal flora disorder caused by ampicillin, and promoting the production of short-chain fatty acids and maintaining the intestinal barrier.

[0024] (2) The prebiotic composition for proliferating beneficial intestinal flora provided by the present invention creatively combines guar gum and flaxseed and then performs specific enzymatic hydrolysis. The obtained partially hydrolyzed composite gum provides a prebiotic substrate with more comprehensive functionality and higher nutritional value for intestinal flora. The partially hydrolyzed composite gum is obtained by integrating guar gum rich in β-mannan and flaxseed gum containing both arabinoxylan and rhamnogalacturonic acid into a composite enzymatic hydrolysis, which 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 fermentable substrate. This structural diversity enables it to be utilized by more beneficial strains with different nutritional preferences, and has a broader spectrum of prebiotic effects than single partially hydrolyzed guar gum.

[0025] (3) The prebiotic composition for proliferating beneficial intestinal flora provided by the present invention obtains a green banana resistant starch (RS2 type) with excellent sustained-release performance through a specific method. Through two-step enzymatic hydrolysis, the natural granular structure of green banana resistant starch is maximized and purified with high efficiency under relatively mild conditions. First, it is enzymatically hydrolyzed by a complex enzyme, gently "breaking the wall" and releasing complete starch granules at low temperature. Then, under elevated temperature conditions, the gelatinized non-resistant starch is degraded by high-temperature resistant α-amylase to obtain high-purity RS2 type resistant starch. Its natural dense semi-crystalline granular structure enables it to effectively resist degradation by gastric acid and small intestinal digestive enzymes and reach the colon intact. Its fermentation process is smooth and long-lasting, allowing it to serve as an energy carrier and successfully cross the proximal and middle parts of the colon, accurately transporting carbon source substances to the distal part of the colon, providing an efficient and impurity-free carbon source for Faecalibacterium prausnitzii and the like, thereby proliferating beneficial flora in the intestine. DETAILED DESCRIPTION

[0026] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0027] Unless otherwise specified, the chemical reagents and materials in the present invention are purchased from commercial sources or synthesized from commercially purchased raw materials.

[0028] The enzyme activity of the β-mannanase is 10,000 U / g, the enzyme activity of the xylanase is 30,000 U / g, the enzyme activity of the polygalacturonase is 60,000 U / g, the enzyme activity of the pectin lyase is 50,000 U / g, the enzyme activity of the cellulase, pectinase and protease is 60,000 U / g, and the enzyme activity of the high-temperature resistant α-amylase is 40,000 U / g.

[0029] Example 1 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 35 parts of green banana resistant starch, 20 parts of enzymatically hydrolyzed okra polysaccharides, and 4 parts of tea polyphenols.

[0030] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 75 g of guar gum powder and 25 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 50° C. for 50 min, and then a 5 wt % citric acid aqueous solution was added to adjust the pH to 4.5, and then 0.4 g of β-mannanase was added, and enzymatic hydrolysis was carried out at 50° C. for 1.5 h. After enzymatic hydrolysis, 0.13 g of xylanase, 0.06 g of polygalacturonase and 0.04 g of pectin lyase were added, and enzymatic hydrolysis was carried out at 50° C. for 2.5 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated in boiling water for 10 min, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25,000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25,000 Da, and the permeate was then passed through a 15,000 Da ultrafiltration membrane to obtain a retentate, and the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0031] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.25% citric acid solution, immersed at 25°C for 13 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 700g of deionized water, quickly pulped, and then 0.4g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5) was added, the pH was adjusted to 5, and enzymatic hydrolysis was carried out at 43°C for 2.5 hours. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.15g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 87°C for 1.5 hours. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0032] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3500g of deionized water, and mashed into a pulp. Subsequently, 0.7g of protease and 0.3g of pectinase were added, and enzymatic hydrolysis was carried out at 50°C and pH 6 for 2.5h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0033] Example 2 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 35 parts of partially hydrolyzed composite gum, 30 parts of green banana resistant starch, 15 parts of enzymatically hydrolyzed okra polysaccharides, and 3 parts of tea polyphenols.

[0034] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 80 g of guar gum powder and 20 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 45 ° C for 60 minutes, and then a 5 wt% citric acid aqueous solution was added to adjust the pH to 4, and then 0.3 g of β-mannanase was added, and enzymatic hydrolysis was carried out at 45 ° C for 2 hours. After enzymatic hydrolysis, 0.1 g of xylanase, 0.05 g of polygalacturonase and 0.05 g of pectin lyase were added, and enzymatic hydrolysis was carried out at 45 ° C for 3 hours. After the enzymatic hydrolysis was completed, the enzyme was inactivated in boiling water for 10 minutes, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25000 Da, and then the permeate was passed through a 15000 Da ultrafiltration membrane to obtain a retentate, and then the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0035] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.2% citric acid solution, immersed at 20°C for 15 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 600g of deionized water, quickly pulped, and then 0.3g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3:2:1) was added, the pH was adjusted to 4.5, and enzymatic hydrolysis was carried out at 40°C for 3h. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.1g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 85°C for 1.5h. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0036] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3000g of deionized water, and mashed into a pulp. Subsequently, 0.6g of protease and 0.2g of pectinase were added, and enzymatic hydrolysis was carried out at 45°C and pH 5.5 for 3h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. Four times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0037] Example 3 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 45 parts of partially hydrolyzed composite gum, 40 parts of green banana resistant starch, 25 parts of enzymatically hydrolyzed okra polysaccharides, and 5 parts of tea polyphenols.

[0038] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 70 g of guar gum powder and 30 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 55° C. for 40 min, and then a 5 wt % citric acid aqueous solution was added to adjust the pH to 5, and then 0.5 g of β-mannanase was added, and enzymatic hydrolysis was carried out at 55° C. for 1 h. After enzymatic hydrolysis, 0.15 g of xylanase, 0.07 g of polygalacturonase and 0.03 g of pectin lyase were added, and enzymatic hydrolysis was carried out at 55° C. for 2 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated in boiling water for 10 min, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25,000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25,000 Da, and the permeate was then passed through a 15,000 Da ultrafiltration membrane to obtain a retentate, and the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0039] The preparation method of the green banana resistant starch is as follows: The green bananas are peeled, the pulp is sliced, and then added to a 0.3% citric acid solution, immersed at 30°C for 10 minutes, then removed, drained, dried, and crushed to obtain green banana powder; 100g of green banana powder is added to 800g of deionized water, quickly pulped, and then 0.5g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 4:3:2) is added, the pH is adjusted to 5.5, and enzymolysis is carried out at 45°C for 2h. After the enzymolysis is completed, it is filtered to obtain green banana starch milk; the green banana starch milk is heated to 85°C, and then 0.2g of high-temperature resistant α-amylase is added, the pH is adjusted to 6, and enzymolysis is carried out at 90°C for 1h. After the enzymolysis is completed, the temperature is lowered to below 60°C, centrifuged at high speed, the precipitate is collected, and the precipitate is washed and dried to obtain the product.

[0040] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200 g of okra pods were washed and cut into segments, added to 4000 g of deionized water, and mashed into a slurry. Subsequently, 0.8 g of protease and 0.4 g of pectinase were added, and enzymatic hydrolysis was carried out at 55°C and pH 6 for 2 h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0041] Comparative Example 1 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 35 parts of green banana resistant starch, 20 parts of enzymatically hydrolyzed okra polysaccharides, and 4 parts of tea polyphenols.

[0042] Wherein, the preparation method of described partial water guar gum is as follows: 100 g of guar gum powder was added to 2 kg of deionized water, heated and stirred at 50 ° C for 50 min, and then a 5 wt% citric acid aqueous solution was added to adjust the pH to 4.5. Then, 0.4 g of β-mannanase was added and enzymolysis was carried out at 50 ° C for 4 h. After the enzymolysis was completed, the enzyme was inactivated in boiling water for 10 min. After cooling to room temperature, the filtrate was passed through a 25000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25000 Da. The permeate was then passed through a 15000 Da ultrafiltration membrane to obtain a retentate. The retentate was then concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0043] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.25% citric acid solution, immersed at 25°C for 13 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 700g of deionized water, quickly pulped, and then 0.4g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5) was added, the pH was adjusted to 5, and enzymatic hydrolysis was carried out at 43°C for 2.5 hours. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.15g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 87°C for 1.5 hours. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0044] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3500g of deionized water, and mashed into a pulp. Subsequently, 0.7g of protease and 0.3g of pectinase were added, and enzymatic hydrolysis was carried out at 50°C and pH 6 for 2.5h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0045] Compared with Example 1, flaxseed gum was not added to the partially hydrolyzed composite gum in this comparative example.

[0046] Comparative Example 2 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 35 parts of green banana resistant starch, 20 parts of enzymatically hydrolyzed okra polysaccharides, and 4 parts of tea polyphenols.

[0047] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 75g of guar gum powder and 25g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2kg of deionized water, heated and stirred at 50°C for 50min, and then a 5wt% citric acid aqueous solution was added to adjust the pH to 4.5, followed by adding 0.4g of β-mannanase, and enzymolysis at 50°C for 2.5h. After enzymolysis, 0.13g of xylanase, 0.06g of polygalacturonase and 0.04g of pectin lyase were added, and enzymolysis was carried out at 50°C for 3.5h. After the enzymolysis was completed, the enzyme was inactivated with boiling water for 10min, cooled to room temperature and filtered, and the filtrate was passed through an ultrafiltration membrane of 15000Da to obtain a permeate with a molecular weight of less than 15000Da, and then the permeate was concentrated under reduced pressure at 15kPa and spray-dried to obtain the product.

[0048] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.25% citric acid solution, immersed at 25°C for 13 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 700g of deionized water, quickly pulped, and then 0.4g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5) was added, the pH was adjusted to 5, and enzymatic hydrolysis was carried out at 43°C for 2.5 hours. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.15g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 87°C for 1.5 hours. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0049] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3500g of deionized water, and mashed into a pulp. Subsequently, 0.7g of protease and 0.3g of pectinase were added, and enzymatic hydrolysis was carried out at 50°C and pH 6 for 2.5h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0050] Compared with Example 1, the molecular weight of the partially hydrolyzed composite glue prepared in this comparative example is less than 15,000 Da.

[0051] Comparative Example 3 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 35 parts of green banana resistant starch, 20 parts of enzymatically hydrolyzed okra polysaccharides, and 4 parts of tea polyphenols.

[0052] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 75 g of guar gum powder and 25 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 50° C. for 50 min, and then a 5 wt % citric acid aqueous solution was added to adjust the pH to 4.5, followed by adding 0.4 g of β-mannanase, and enzymolysis was carried out at 50° C. for 0.5 h. After enzymolysis, 0.13 g of xylanase, 0.06 g of polygalacturonase and 0.04 g of pectin lyase were added, and enzymolysis was carried out at 50° C. for 1.5 h. After the enzymolysis was completed, the enzyme was inactivated with boiling water for 10 min, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25,000 Da ultrafiltration membrane to obtain a retentate with a molecular weight greater than 25,000 Da, and the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0053] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.25% citric acid solution, immersed at 25°C for 13 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 700g of deionized water, quickly pulped, and then 0.4g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5) was added, the pH was adjusted to 5, and enzymatic hydrolysis was carried out at 43°C for 2.5 hours. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.15g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 87°C for 1.5 hours. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0054] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3500g of deionized water, and mashed into a pulp. Subsequently, 0.7g of protease and 0.3g of pectinase were added, and enzymatic hydrolysis was carried out at 50°C and pH 6 for 2.5h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0055] Compared with Example 1, the molecular weight of the partially hydrolyzed composite glue prepared in this comparative example is greater than 25,000 Da.

[0056] Comparative Example 4 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 20 parts of enzymatically hydrolyzed okra polysaccharide, and 4 parts of tea polyphenols.

[0057] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 75 g of guar gum powder and 25 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 50° C. for 50 min, and then a 5 wt % citric acid aqueous solution was added to adjust the pH to 4.5, and then 0.5 g of β-mannanase was added, and enzymatic hydrolysis was carried out at 50° C. for 1.5 h. After enzymatic hydrolysis, 0.13 g of xylanase, 0.06 g of polygalacturonase and 0.04 g of pectin lyase were added, and enzymatic hydrolysis was carried out at 50° C. for 2.5 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated in boiling water for 10 min, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25,000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25,000 Da, and the permeate was then passed through a 15,000 Da ultrafiltration membrane to obtain a retentate, and the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0058] The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: 200g of okra pods were washed and cut into segments, added to 3500g of deionized water, and mashed into a pulp. Subsequently, 0.7g of protease and 0.3g of pectinase were added, and enzymatic hydrolysis was carried out at 50°C and pH 6 for 2.5h. After the enzymatic hydrolysis was completed, the supernatant was taken and concentrated under reduced pressure to 1 / 4 of the original volume. 4 times the volume of anhydrous ethanol was added for alcohol precipitation. The precipitate was washed and vacuum-dried to obtain enzymatically hydrolyzed okra polysaccharide.

[0059] Compared with Example 1, the prebiotic composition in this comparative example does not contain green banana resistant starch.

[0060] Comparative Example 5 A prebiotic composition for proliferating beneficial intestinal bacteria, comprising the following raw materials in parts by weight: 40 parts of partially hydrolyzed composite gum, 35 parts of green banana resistant starch, and 4 parts of tea polyphenols.

[0061] Wherein, the preparation method of the partially hydrolyzed composite glue is as follows: 75 g of guar gum powder and 25 g of linseed gum powder were fully mixed to obtain composite gum powder, and then the composite gum powder was added to 2 kg of deionized water, heated and stirred at 50° C. for 50 min, and then a 5 wt % citric acid aqueous solution was added to adjust the pH to 4.5, and then 0.5 g of β-mannanase was added, and enzymatic hydrolysis was carried out at 50° C. for 1.5 h. After enzymatic hydrolysis, 0.13 g of xylanase, 0.06 g of polygalacturonase and 0.04 g of pectin lyase were added, and enzymatic hydrolysis was carried out at 50° C. for 2.5 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated in boiling water for 10 min, and the mixture was cooled to room temperature and filtered. The filtrate was passed through a 25,000 Da ultrafiltration membrane to obtain a permeate with a molecular weight of less than 25,000 Da, and the permeate was then passed through a 15,000 Da ultrafiltration membrane to obtain a retentate, and the retentate was concentrated under reduced pressure at 15 kPa and spray-dried to obtain the product.

[0062] The preparation method of the green banana resistant starch is as follows: The green bananas were peeled, the pulp was sliced, and then added to a 0.25% citric acid solution, immersed at 25°C for 13 minutes, then removed and drained, dried, and crushed to obtain green banana powder; 100g of green banana powder was added to 700g of deionized water, quickly pulped, and then 0.4g of complex enzyme (cellulase, pectinase, and protease in a mass ratio of 3.5:2.5:1.5) was added, the pH was adjusted to 5, and enzymatic hydrolysis was carried out at 43°C for 2.5 hours. After the enzymatic hydrolysis was completed, it was filtered to obtain green banana starch milk; the green banana starch milk was heated to 85°C, and then 0.15g of high-temperature resistant α-amylase was added, the pH was adjusted to 6, and enzymatic hydrolysis was carried out at 87°C for 1.5 hours. After the enzymatic hydrolysis was completed, the temperature was lowered to below 60°C, and high-speed centrifugation was performed to collect the precipitate, which was washed and dried to obtain the product.

[0063] Compared with Example 1, the prebiotic composition in this comparative example does not contain enzymatically hydrolyzed okra polysaccharide.

[0064] The prebiotic compositions prepared in Examples 1-3 and Comparative Examples 1-5 were subjected to mouse experiments, as follows: One hundred SPF-grade male BALB / c mice were randomly divided into 10 groups of 10 mice each. BALB / c mice, 6–8 weeks old and weighing 18–22 g, were purchased from Chongqing Tengxin Biotechnology Co., Ltd. The experimental animals were housed in a barrier animal room at 22°C, 40–70% humidity, and a 12-h light–dark cycle. They were fed a standard diet and acclimated for 1 week with free access to water. The blank control group was gavaged with an equal volume of normal saline. The remaining groups were first treated with ampicillin to disrupt the bacterial flora. The treatment was then continued by gavage at a dose of 300 μL of ampicillin (1 mg / mL) once daily for 14 days. After the model was established, the prebiotic composition in each group was prepared into a 25 mg / mL solution. The Example 1-3 groups were then gavaged with the prebiotic composition of Examples 1-3, and the Comparative Example 1-5 groups were gavaged with the prebiotic composition of Comparative Example 1-5. The gavage dose for each group was 0.4 mL (500 mg / kg bw). The blank control and model groups were gavaged with an equal volume of normal saline. The test samples were administered for 30 days. The initial fasting body weight of the mice and the fasting body weight at the end of the experiment were recorded, as shown in Table 1.

[0065] Table 1 Body weight changes of mice As shown in Table 1 above, the model group showed significantly lower weight gain compared to the blank group, indicating successful model establishment. The weight gain of mice in Examples 1-3 was significantly higher than that in the model group, demonstrating that the prebiotic composition prepared by the present invention can significantly promote weight recovery and growth in mice. Furthermore, the performance of Examples 1-3 was superior to that of Comparative Examples 1-5, demonstrating a synergistic effect between the ingredients in the prebiotic composition of the present invention.

[0066] 24 hours after oral administration of the prebiotic composition on day 0 and day 30, 0.1 g of feces from the anus of mice in the blank group, model group, Example 1 group, and Comparative Example 1-5 groups were collected aseptically, placed in a dry sterilized test tube, and diluted to 10% by adding sterile diluent. -6 , respectively inoculated onto bifidobacterium, lactobacillus, enterobacter, enterococcus, Clostridium perfringens, and Clostridium prausnitzii culture media, wherein the bifidobacterium culture medium is BBL agar, the lactobacillus culture medium is LBS agar, the enterobacterium culture medium is eosin methylene blue agar, the enterococcus culture medium is sodium azide-crystal violet-esculin agar, the Clostridium perfringens culture medium is TSC agar, and the Clostridium prausnitzii culture medium is YCFA liquid culture medium. The culture was carried out in an anaerobic environment for 48 hours, and 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.

[0067] Table 2 Effects of each group of prebiotic compositions on intestinal flora of mice ( ±s, log cfu / g, n=10) Note: # p<0.05, compared with the blank 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.

[0068] As can be seen from Table 2 above, compared with the blank group, in the model group mouse feces, enterococci, enterobacter and Clostridium perfringens significantly increase, while lactobacillus, bifidobacterium, Clostridium prausnitzii significantly reduce, illustrating that ampicillin has changed the mouse intestinal flora composition, and film making is successful. Compared with the model group, in the mouse feces flora of embodiment 1 group feeding, bifidobacterium, lactobacillus content and Clostridium prausnitzii significantly increase, and Clostridium perfringens significantly reduces, illustrating that the prebiotic composition prepared by the present invention increases the content of beneficial bacteria, while enterococci, enterobacterium are returned to normal flora content, and there is a significant repair effect for intestinal flora imbalance, maintaining the balance of intestinal microbial group. And embodiment 1 group has the content that significantly improves bifidobacterium, lactobacillus content and Clostridium prausnitzii compared to comparative example 1-5 group, illustrating that there is synergism between the raw material in the prebiotic composition of the present invention.

[0069] The prebiotic compositions prepared in Example 1 and Comparative Examples 1-5 were subjected to in vitro fermentation tests to measure changes in gas production, as follows: Preparation of basal anaerobic 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), resazurin solution (1.0 mL, 1%, w / v), vitamin K (10.0 μL) and ultrapure water. The pH of the basal nutrient medium was adjusted to pH = 7.0 and sterilized by high-pressure steam at 121°C for 15 min.

[0070] Preparation of phosphate buffered saline (PBS): Sodium chloride (2.0 g), potassium dihydrogen phosphate (0.2 g), disodium hydrogen phosphate (2.9 g), and potassium chloride (0.2 g) were dissolved in 1 L of ultrapure water, sterilized and cooled to adjust the pH to 7.4, and refrigerated at 4°C until use.

[0071] On the day of the experiment, feces were collected from healthy adults (1 male and 1 female) (healthy, without gastrointestinal diseases and without taking antibiotics in the past three months). The collected fecal samples were transferred to an anaerobic incubator for processing. 10 g of each fecal sample was weighed on an electronic balance and mixed with phosphate buffer solution at a ratio of 1:10 (W / V) (one male and one female was used to obtain a richer flora to construct an intestinal flora model), and filtered through gauze for later use.

[0072] Six Erlenmeyer flasks were each filled with 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. An equal amount of sterile water was used in place of the prebiotic composition in the blank control group. The cells were incubated in a 37°C anaerobic incubator. Gas production in each group was measured using a graduated syringe at 4, 8, 12, 16, 20, and 24 hours of fermentation. The results are shown in Table 3. Table 3 Effects of different prebiotic compositions on gas production (mL) 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 (4-12 hours) and a continuous and stable gas production in the later stage (12-24 hours), which can slow the gas production rate during fermentation in the intestine and reduce the discomfort caused by abdominal distension. In contrast, in Comparative Example 2, due to the decrease in molecular weight of the partially hydrolyzed composite gum and the lack of green banana resistant starch in Comparative Example 4, the gas production rate in the early stage is significantly increased, resulting in the production of large amounts of gas, which can easily lead to obvious intestinal intolerance symptoms such as abdominal distension, bowel sounds, and increased flatus.

[0073] The above content is a further detailed description of the present invention in combination with specific implementation examples. It cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, they can make several simple deductions or substitutions without departing from the concept of the present invention, which should be regarded as falling within the scope of protection of the present invention.

[0074] It will be easily understood by those skilled in the art 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 in the scope of protection of the present invention.

Claims

1. A prebiotic composition for proliferating beneficial intestinal bacteria, characterized in that: Calculated by weight, it includes the following raw materials: 35-45 parts of partially hydrolyzed composite gum, 30-40 parts of green banana resistant starch, 15-25 parts of enzymatically hydrolyzed okra polysaccharide, and 3-5 parts of tea polyphenols.

2. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 1, characterized in that: Calculated by weight, it includes the following raw materials: 40-45 parts of partially hydrolyzed composite gum, 30-35 parts of green banana resistant starch, 20-25 parts of enzymatically hydrolyzed okra polysaccharide, and 4-5 parts of tea polyphenols.

3. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 1, characterized in that: The partially hydrolyzed composite gum is obtained by mixing guar gum and flaxseed gum and then performing enzymatic hydrolysis, and has a molecular weight of 15,000-25,000 Da.

4. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 1, characterized in that: The preparation method of the partially hydrolyzed composite glue is as follows: Guar gum powder and flaxseed gum powder are fully mixed to obtain composite gum powder, and then the composite gum powder is added to deionized water, heated and stirred, and then citric acid is added to adjust the pH, and then β-mannanase is added to perform a primary enzymolysis. After the enzymolysis reaches a predetermined time, xylanase, polygalacturonase and pectin lyase are added to perform a secondary enzymolysis. After the enzymolysis is completed, the enzyme is inactivated with boiling water, and the mixture is filtered after cooling to room temperature. The filtrate is ultrafiltered, concentrated under reduced pressure, and spray-dried to obtain the product.

5. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 4, characterized in that: The mass ratio of the guar gum powder to the flaxseed gum powder is 7-8:2-3, the heating and stirring temperature is 45-55°C, the time is 40-60 minutes, and the pH is adjusted to 4-5; the amount of the β-mannanase added is 0.3-0.5% of the mass of the composite rubber powder, the temperature of the primary enzymatic hydrolysis is 45-55°C, and the time is 1-2 hours; the amount of the xylanase added is 0.1-0.15% of the mass of the composite rubber powder, the amount of the polygalacturonase added is 0.05-0.07% of the mass of the composite rubber powder, the amount of the pectin lyase added is 0.03-0.05% of the mass of the composite rubber powder, and the temperature of the secondary enzymatic hydrolysis is 45-55°C, and the time is 2-3 hours.

6. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 1, characterized in that: The preparation method of the green banana resistant starch is as follows: The green bananas are peeled, the pulp is sliced, and then immersed in a citric acid solution, then removed, drained, dried, and crushed to obtain green banana powder; the green banana powder is added to deionized water, quickly pulped, and then a complex enzyme is added to adjust the pH to perform a primary enzymatic hydrolysis, and filtered after the enzymatic hydrolysis is completed to obtain green banana starch milk; the green banana starch milk is heated, and then a high-temperature resistant α-amylase is added to perform a secondary enzymatic hydrolysis, and after the enzymatic hydrolysis is completed, the temperature is cooled to below 60°C, and high-speed centrifugation is performed to collect the precipitate, and the precipitate is washed and dried to obtain the product.

7. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 6, characterized in that: The mass concentration of the citric acid solution is 0.2-0.3%, the immersion temperature is 20-30°C, and the immersion time is 10-15 minutes; the material-liquid ratio of the green banana powder to deionized water is 1:6-8, the added amount of the complex enzyme is 0.3-0.5% of the mass of the green banana powder, and the complex enzyme is composed of cellulase, pectinase, and protease in a mass ratio of 3-4:2-3:1-2. The temperature of the primary enzymatic hydrolysis is 40-45°C, the pH is 4.5-5.5, and the time is 2-3 hours; the added amount of the high-temperature resistant α-amylase is 0.1-0.2% of the mass of the green banana powder, and the temperature of the secondary enzymatic hydrolysis is 85-90°C, and the time is 1-1.5 hours.

8. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 1, characterized in that: The preparation method of the enzymatic hydrolysis of okra polysaccharide is as follows: The okra pods are cleaned and cut into segments, added into deionized water, and mashed into a pulp. Subsequently, protease and pectinase are added for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, the mixture is centrifuged, the supernatant is taken, 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 vacuum dried to obtain enzymatically hydrolyzed okra polysaccharides.

9. The prebiotic composition for proliferating beneficial intestinal bacteria according to claim 8, characterized in that: The mass ratio of the okra pods to deionized water is 1:15-20, the added amount of the protease is 0.3-0.4% of the mass of the okra pods, the added amount of the pectinase is 0.1-0.2% of the mass of the okra pods, the temperature of the enzymolysis is 45-55°C, the pH is 5.5-6, and the time is 2-3h.

10. A method for preparing a prebiotic composition for proliferating beneficial intestinal bacteria according to any one of claims 1 to 9, characterized in that: The following steps are involved: Weigh the raw materials according to the formula and mix them evenly.

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