A functional food composition for regulating intestinal flora in children and a method for preparing the same

Abstract

This invention discloses a functional food composition for regulating children's intestinal flora and its preparation method, relating to the field of functional food processing technology. The composition includes a complete fermentation product obtained through layered solid-state anaerobic in-situ induced fermentation. The solid substrate used for fermentation, by weight, includes 28-38 parts fermented yam powder, 18-25 parts hawthorn residue, 10-16 parts blueberry residue, 12-18 parts Poria cocos powder, and 4-9 parts xylooligosaccharides. The fermentation process uses mulberry and tangerine peel extract as an exogenous signal substrate, and the inoculated microorganisms include Bacillus subtilis, Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei. This invention relies on layered solid-state anaerobic in-situ fermentation and stepwise inoculation technology, combined with timed feeding of mulberry and tangerine peel compound extract, and freeze-dried and ultra-finely pulverized into chewable tablets. This efficiently produces active postbiotics, utilizes all raw materials, is suitable for children's intestines, inhibits harmful bacteria, and improves digestive capacity.

Description

Technical Field

[0001] This invention relates to the field of functional food processing technology, specifically to a functional food composition for regulating children's intestinal flora and its preparation method. Background Technology

[0002] The homeostasis of a child's gut microbiota directly determines their digestive and absorptive capacity, immune development level, and daily growth and development. Insufficient beneficial bacteria and excessive proliferation of harmful bacteria in the gut can easily lead to various common digestive problems such as bloating, indigestion, irregular bowel movements, and loss of appetite. Currently, food products on the market for regulating gut health are mainly divided into three categories: single probiotic preparations, ordinary prebiotic powders, and dietary supplements made from simple compound medicinal and edible ingredients. Probiotic products have drawbacks such as poor stability of live bacteria during storage and transportation, and susceptibility to inactivation by gastric acid and bile after entering the digestive tract. Simple prebiotics can only provide nutrition to the inherent gut microbiota and cannot... Actively inducing the generation of beneficial metabolites; ordinary food and medicine homology compound products mostly adopt simple mixing and processing technology, and the large molecules such as starch, dietary fiber, and polyphenols in the raw materials are difficult for the human body to absorb directly. Solid-state fermentation technology is now widely used in the field of deep processing of functional raw materials. With the help of microbial enzymatic hydrolysis, large molecular substrates can be decomposed to generate short-chain fatty acids, bacteriocins and other metabiotics with intestinal conditioning effects. Layered solid-state fermentation, multi-strain stepwise fermentation and exogenous signal-induced fermentation are gradually becoming the mainstream directions of functional food research and development, providing a feasible technical path for developing intestinal conditioning foods that are suitable for children's physiological characteristics.

[0003] Existing solid-state fermentation processes for preparing intestinal conditioning foods suffer from several unavoidable drawbacks. Conventional fermentation systems employ a homogeneous, single-layer mixing method, where all raw materials are mixed and fermented simultaneously. This fails to differentiate between microbial growth environments suited to aerobic and anaerobic metabolism. Simultaneous inoculation of Bacillus and Lactobacillus leads to competition among microbial species, limiting the proliferation of various microorganisms and resulting in low yields of active metabolites. Furthermore, traditional fermentation processes lack exogenous inducing signaling substances, relying solely on the inherent nutrients of the substrate for microbial metabolism. This hinders the continuous and targeted synthesis of functional metabolites that improve the intestinal barrier. Most processing steps... After fermentation, the fermentation residue is discarded, and only a small amount of liquid active ingredients are extracted. A large amount of dietary fiber and medicinal and edible functional components are wasted, resulting in low overall utilization efficiency of raw materials. The drying process generally uses hot air drying, and the high temperature environment will destroy heat-sensitive active substances such as short-chain fatty acids and bacteriocins, which will significantly weaken the actual efficacy of the finished product in regulating the intestines. A large amount of filler and auxiliary materials are added to the molded products to reduce the proportion of active base materials. Some of these auxiliary materials can easily irritate the delicate stomachs of children. At the same time, the single fermentation environment is poorly controlled, the oxygen content inside the fermentation bed cannot be precisely controlled, the probability of contamination by miscellaneous bacteria is high, and the storage stability of the finished product is poor. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a functional food composition for regulating children's intestinal flora and its preparation method. It uses medicinal and edible materials such as yam, hawthorn residue, blueberry residue, and Poria cocos as solid-state fermentation substrates, combined with xylooligosaccharides to construct a nutrient matrix, and mulberry and tangerine peel extract as an exogenous inducing signal. Multiple types of probiotics are selected for staged inoculation and fermentation. This invention sets up a three-layered fermentation matrix. First, aerobic pre-fermentation utilizes Bacillus subtilis to degrade macromolecules. Then, nitrogen is replaced to create an anaerobic environment, driving the metabolism of compound lactic acid bacteria to produce short-chain fatty acids, bacteriocins, and other post-biotic active ingredients. After fermentation, all materials are mixed, freeze-dried, and ultra-finely pulverized to obtain the base material. A small amount of food excipients is then added to make chewable tablets. The entire process achieves full utilization of the substrate components, accurately distinguishes between aerobic and anaerobic fermentation environments, reduces strain competition, and increases the production of active substances. The finished product is mild and suitable for children's intestines, balancing intestinal flora and improving digestion. The production process has low contamination risk and strong storage stability, making it suitable for large-scale production as a daily intestinal conditioning dietary food for children.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution: On the one hand, a functional food composition for regulating children's intestinal flora, the composition comprising a complete fermentation product obtained by layered solid-state anaerobic in-situ induced fermentation; The solid substrate used for fermentation includes, by weight, 28-38 parts fermented yam powder, 18-25 parts hawthorn residue, 10-16 parts blueberry pomace, 12-18 parts Poria cocos powder, and 4-9 parts xylooligosaccharides. The fermentation process uses mulberry and tangerine peel induction extract as an exogenous signal substrate. The microbial community used in the fermentation includes Bacillus subtilis, Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei. The microbial community is added in a stepwise inoculation manner. The fermentation system consists of a three-layer matrix structure, which, from bottom to top, consists of a bottom layer of mixed hawthorn residue and blueberry residue, a middle layer of mixed fermented yam powder, poria cocos fragments, and xylooligosaccharides, and a top layer of bran as an isolation layer.

[0006] Furthermore, the fermented yam powder is made from fresh yam through washing, slicing, steaming, drying, and ultra-fine grinding. The thickness of the yam slices is 3mm to 6mm, the steaming time is 25min to 40min, the drying temperature is 50℃ to 55℃, and the moisture content of the dried material is less than 10%. The hawthorn pomace and blueberry pomace are by-products of juice extraction. After being washed and drained, the two types of pomace are crushed to 20-40 mesh. The Poria cocos powder is obtained by pulverizing Poria cocos raw material after washing and drying. The mulberry and tangerine peel induced extract is obtained by mixing mulberry and tangerine peel at a mass ratio of 3:1 to 5:1, adding purified water, and extracting at 55℃ to 65℃ for 2 to 3 hours, followed by filtration to remove solid residue.

[0007] Furthermore, after the three-layer matrix is ​​laid, the moisture content of the bottom layer of hawthorn residue and blueberry residue mixture is adjusted to 48%, and half of the total amount of mulberry and tangerine peel induced extract is sprayed onto the bottom layer. The moisture content of the mixture of medium-fermented yam powder, chopped Poria cocos, and xylooligosaccharides is adjusted to 32%. The top layer of bran separator should be 0.5cm to 1cm thick, and the moisture content of the separator should be adjusted to 20%. The thickness of the bottom layer is 6cm to 10cm, and the thickness of the middle layer is 8cm to 12cm.

[0008] Furthermore, the Bacillus subtilis is inoculated into the middle layer substrate during the aerobic pre-fermentation enzymatic hydrolysis activation process, with the pre-fermentation temperature at a constant 32℃~36℃ and a total duration of 22h~26h; Sterile air was introduced every 6 hours during the pre-fermentation cycle, with a single aeration time of 8 to 12 minutes, and the oxygen volume fraction in the bed was maintained at 8% to 12%. A mixed culture of Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei was inoculated into the middle and bottom substrates after pre-fermentation.

[0009] Furthermore, after the lactic acid bacteria mixed bacterial solution is introduced, the air inside the fermentation bed cavity is replaced three times with sterile nitrogen gas, and a trace amount of sterile nitrogen gas is continuously introduced to maintain a positive pressure anaerobic environment in the cavity, controlling the oxygen volume fraction of the bed to be below 2.0%; the anaerobic fermentation temperature range is 31℃~33℃, and the anaerobic fermentation duration is 72h~96h; during the anaerobic fermentation stage, lactose solution and the remaining mulberry and tangerine peel induction extract are added to the bottom substrate every 24h, and the total mass of the materials added at one time is 1%~2% of the dry weight of the bottom substrate.

[0010] Furthermore, the water activity of the whole-component fermentation product is no greater than 0.6, and the whole-component fermentation product contains short-chain fatty acids and bacteriocins produced by the fermentation process.

[0011] Furthermore, after all fermentation processes are completed, the bottom, middle, and top layers of materials are thoroughly mixed, and the mixture is then vacuum freeze-dried to remove free moisture. The moisture content of the dried material is controlled to be no more than 5%. The dried material is pulverized and sieved to obtain a composite post-genetic matrix powder of 80-100 mesh.

[0012] Furthermore, the composite post-biotic base powder is mixed evenly with food-grade excipients and then fed into a rotary tablet press to be pressed into chewable tablets; The base powder accounts for 95% to 98% of the total mass of the chewable tablets, and food-grade excipients account for 2% to 5%. Food-grade excipients include microcrystalline cellulose, sorbitol, and magnesium stearate.

[0013] On the other hand, a functional food composition for regulating children's gut microbiota and its preparation method are disclosed, the specific steps of which are as follows: S1, complete the standardized pretreatment of fermented yam powder, hawthorn residue, blueberry residue and poria cocos powder, and simultaneously complete the extraction and solid-liquid separation of mulberry and tangerine peel induced extract, and collect all solid raw materials and liquid induced extract for later use. S2, In the solid fermentation bed chamber, the bottom layer material, the middle layer material and the top layer bran isolation layer are laid from bottom to top in sequence, and the moisture content of each layer of material is adjusted layer by layer to seal the fermentation bed chamber; S3, the Bacillus subtilis bacterial solution that has been activated step by step is evenly sprayed into the middle layer of the substrate, the middle layer material is stirred and mixed, the fermentation bed is sealed and a constant temperature is set, and sterile air is intermittently introduced to maintain a specific oxygen concentration in the chamber for constant temperature aerobic pre-fermentation. S4. After pre-fermentation, stop the introduction of sterile air. Spray the activated Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei mixed bacterial solution onto the middle and bottom substrates and stir the materials. Introduce sterile nitrogen to replace the air inside the chamber. Continuously introduce a small amount of sterile nitrogen to maintain a positive pressure anaerobic environment inside the chamber. Set a constant fermentation temperature for anaerobic fermentation. During the anaerobic fermentation cycle, add lactose solution and the remaining mulberry and tangerine peel induction extract to the bottom substrate at regular intervals. The total mass of each addition is 1% to 2% of the dry weight of the bottom substrate. S5. After the anaerobic induced co-fermentation process is completed, the temperature control parameters of the fermentation bed are lowered to keep the fermentation bed in a closed state and continue to be statically fermented for post-ripening conversion. S6. After the post-ripening conversion is completed, the fermentation bed is turned on, and the bottom, middle and top layers of materials are fully mixed to obtain mixed fermentation material. The mixed fermentation material is sent to a vacuum freeze dryer for dehydration treatment. The dried material is then pulverized and sieved to obtain base powder. The base powder is fully mixed with food-grade excipients, and the mixed powder is sent to a rotary tablet press to be pressed into chewable tablets.

[0014] Compared with existing technologies, this functional food composition for regulating children's gut microbiota and its preparation method have the following beneficial effects: I. This invention utilizes a layered solid-state anaerobic in-situ induced fermentation system combined with a multi-strain stepwise inoculation process. Relying on differentiated material ratios and layered water control in the layered substrate, it constructs a gradient nutrient and permeable anaerobic zone environment. Different substrate layers are adapted to the needs of aerobic pre-fermentation and deep anaerobic fermentation, respectively. First, Bacillus subtilis is used to complete the enzymatic degradation of substrate macromolecules, and then compound lactic acid bacteria are used to achieve anaerobic metabolic transformation. The layered isolation layer can prevent the entry of external bacteria while locking in the active signal substances of the bottom-layer induced extract, preventing the volatilization and loss of active ingredients. Multi-stage differentiated oxygen environment control can adapt to the metabolic characteristics of different bacterial species, allowing various probiotics to proliferate in an orderly manner and continuously metabolize to produce short-chain fatty acids, bacteriocins, and other active postbiotic substances. The resulting fermentation product completely retains all functional components of the substrate, eliminating the need for separation and waste. It fully leverages the synergistic conditioning effect of medicinal and edible homologous raw materials, gently adapting to the immature intestinal environment of children, regulating the balance of intestinal flora structure, inhibiting the colonization of harmful bacteria, improving the integrity of the intestinal barrier, significantly increasing raw material utilization, and significantly improving the efficiency of active substance production in the fermentation product compared to conventional single solid-state fermentation modes.

[0015] II. This invention utilizes a compound extract of mulberry and tangerine peel as an exogenous signaling substrate, coupled with timed feeding operations, to continuously provide metabolic induction signals and carbon and nitrogen nutrition to the anaerobic fermentation microbial community, promoting the continuous synthesis of functional metabolites by probiotics. Subsequent vacuum freeze-drying combined with ultra-fine pulverization maximizes the retention of heat-sensitive active substances in the fermentation products, avoiding the inactivation of short-chain fatty acids and bacteriocins caused by high-temperature drying. The refined base powder is more easily absorbed and utilized by children's digestive system. A high proportion of fermentation base material is combined with a small amount of mild food excipients to make chewable tablets, eliminating irritating excipients, making the taste suitable for children's eating habits, convenient to take and without burdening the digestive system. The entire preparation process achieves full resource utilization of components, with no processing waste generated. Precise oxygen and nitrogen control during the production process reduces the risk of contamination by miscellaneous bacteria. The water activity of the finished product is controlled within an appropriate range, extending the storage period. Long-term consumption can stably improve children's intestinal digestive and absorptive capacity, reducing problems such as bloating and indigestion caused by intestinal disorders, thus balancing production economy and food safety.

[0016] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0018] Figure 1 Overall process flow diagram of the preparation method of functional food composition for regulating children's intestinal flora; Figure 2 A detailed flowchart of the S2 layered material laying process; Figure 3 A detailed flowchart of the S3 sub-process for aerobic pre-fermentation of Bacillus subtilis. Detailed Implementation

[0019] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below. Example 1:

[0020] Complete manufacturing process of functional food chewable tablets for regulating children's gut microbiota, such as... Figure 1 As shown.

[0021] To prepare fermented yam powder, fresh yams are selected and repeatedly rinsed with clean water to remove surface mud and impurities. The yams are then cut into 4mm thick slices and steamed at a constant temperature for 32 minutes. After steaming, the slices are laid flat in a hot air drying oven at a constant temperature of 52℃ for continuous drying. After drying, the moisture content of the material is measured to be 9.2%. The dried yam slices are then pulverized in an ultra-fine pulverizer and sieved through a 60-mesh screen. The material passing through the screen is collected to obtain fermented yam powder for later use.

[0022] Pre-treatment of hawthorn pomace and blueberry pomace: Hawthorn pomace and blueberry pomace produced by fruit and vegetable juicing are taken, rinsed twice with clean water, placed on a draining table to drain the surface water, and then sent to a pulverizer to be pulverized and sieved to obtain 30-mesh mixed fruit pomace material for later use.

[0023] To prepare Poria cocos powder, wash the whole Poria cocos raw material with clean water, place it in a 50℃ drying equipment to dry it until the moisture content of the material is 8.5%, crush the dried Poria cocos pieces and pass them through a 40-mesh sieve, collect the material that passes through the sieve to obtain Poria cocos powder for later use.

[0024] Preparation of mulberry and tangerine peel induced extract: Mulberry and tangerine peel were weighed and mixed at a mass ratio of 4:1. All raw materials were put into the extraction tank, and purified water was added to completely submerge the solid materials. The extraction tank was kept at a constant temperature of 60℃ for 2.5 hours. After soaking, the solid residue was removed by filtering with a 200-mesh filter cloth. The clear liquid was collected as mulberry and tangerine peel induced extract and stored in a sealed container at low temperature for later use.

[0025] The fermentation material is layered inside the solid anaerobic fermentation bed from bottom to top: The bottom material consists of 22 parts hawthorn residue and 13 parts blueberry residue. The two are mixed evenly and used as the bottom material. The bottom layer is 8cm thick. Half of the total amount of mulberry and tangerine peel extract is sprayed evenly onto the bottom material. During the spraying process, all the bottom material is stirred at the same time to adjust the overall moisture content of the bottom material to 48%.

[0026] The middle layer consists of 32 parts fermented yam powder, 15 parts crushed Poria cocos, and 6 parts xylooligosaccharide. After mixing the three evenly, spread them on top of the bottom layer. The middle layer is 10cm thick. Spray purified water in small amounts multiple times while simultaneously turning the material over to adjust the moisture content of the middle layer to 32%.

[0027] The top layer of bran, acting as an isolation layer, is laid flat on top of the middle layer of material, with a thickness of 0.8 cm. A small amount of water is added via mist spray, and the bran is stirred. The moisture content of the isolation layer is adjusted to 20%. After all three layers of substrate are laid, the fermentation bed is completely sealed. Figure 2 As shown.

[0028] Bacillus subtilis was activated stepwise using three stages of liquid culture medium to obtain a viable bacterial concentration of 1×10⁻⁶. 9 The bacterial solution was sprayed evenly into the middle layer substrate at a concentration of CFU / mL. The middle layer material was stirred throughout the spraying process to ensure uniform distribution of the bacterial solution. After spraying, the fermentation bed was sealed and pre-fermented at a constant temperature of 34℃ for a total of 24 hours.

[0029] After the pre-fermentation starts, sterile air is introduced every 6 hours, with each aeration lasting 10 minutes. Oxygen concentration is monitored in real-time using a supporting oxygen concentration monitoring device to maintain the bed oxygen volume fraction at 9% to 11%. Sterile air supply is stopped after 24 hours of pre-fermentation. Figure 3 As shown.

[0030] Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei were activated and cultured separately, and then mixed with each other at a 1:1:1 ratio to obtain a mixed lactic acid bacteria solution. The total viable bacteria concentration of the mixed solution was 2 × 10⁻⁶. 9 CFU / mL, spray the mixed bacterial solution evenly onto the middle and bottom substrates, and stir the materials layer by layer to ensure that the bacterial solution fully contacts the solid substrate.

[0031] Open the nitrogen pipeline to introduce sterile nitrogen to replace the air inside the chamber. Repeat the complete replacement operation 3 times. After the replacement is completed, continuously introduce a small amount of sterile nitrogen to maintain a slight positive pressure in the chamber. Monitor the oxygen volume fraction of the bed in real time to ensure it is consistently below 1.8%.

[0032] During the anaerobic fermentation stage, the temperature inside the chamber was kept constant at 32℃, and the continuous anaerobic fermentation time was 84h. Every 24h during the anaerobic fermentation process, a lactose aqueous solution was prepared and mixed with the remaining half of the mulberry and tangerine peel induction extract. The mixed liquid was evenly sprayed onto the bottom hawthorn and blueberry pomace substrate. The total mass of the mixed materials added at one time was 1.5% of the dry weight of the bottom substrate.

[0033] After all the anaerobic fermentation processes are completed, keep the fermentation bed sealed without ventilation, and adjust the temperature control parameters of the fermentation bed to 28℃ and let it stand for 12 hours for post-ripening and conversion.

[0034] After the post-ripening conversion is completed, the fermentation bed door is opened, and all materials in the bottom, middle and top layers are thoroughly mixed to obtain a complete fermentation mixture. The mixture is then sent to a vacuum freeze dryer for dehydration. The vacuum level is kept stable at 8 Pa throughout the freeze drying process. After drying, the moisture content of the material is measured to be 4.1%.

[0035] The dried solid was fed into an ultra-fine pulverizer for crushing and then sieved through two stages of 80-mesh and 100-mesh screens. The material under the 100-mesh screen was collected to obtain the composite bio-based powder. The water activity of the powder was measured to be 0.54, and short-chain fatty acids and bacteriocin active substances could be stably detected in the sample.

[0036] Weigh 96 parts of the compound post-generic base powder and mix it with 4 parts of food-grade excipients. The excipient components include 2.2 parts of microcrystalline cellulose, 1.5 parts of sorbitol, and 0.3 parts of magnesium stearate.

[0037] All powders are fed into a three-dimensional motion mixer and mixed at a constant speed for 30 minutes. The uniformly mixed powder is then conveyed to a rotary tablet press, where the tableting pressure is kept constant at 20 kN. The tablets are then stamped to obtain the finished product of functional chewable tablets for regulating children's intestinal flora.

[0038] Finished product basic performance testing: Each tablet weighs 0.8 grams, is intact and unbroken, has a sweet and sour taste without any bitter or irritating odor, and contains a total short-chain fatty acid content of 12.6 mg / g in the base powder. The bacteriocin in the product exhibits an inhibition zone diameter of 11.2 mm against Escherichia coli. Example 2:

[0039] Validation of the optimized weight ratio of solid-state fermentation substrate.

[0040] Three different solid substrate weight fraction gradient settings: Three independent fermentation test groups were set up. All process parameters for pretreatment, fermentation, drying and tableting were kept completely consistent for the three groups, and only the weight ratio of solid substrate was adjusted.

[0041] The first substrate ratio consists of 28 parts fermented yam powder, 18 parts hawthorn residue, 10 parts blueberry pulp, 12 parts Poria cocos powder, and 4 parts xylooligosaccharides.

[0042] The second substrate composition consisted of 33 parts fermented yam powder, 21 parts hawthorn residue, 13 parts blueberry pulp, 15 parts Poria cocos powder, and 6 parts xylooligosaccharides.

[0043] The third substrate composition consisted of 38 parts fermented yam powder, 25 parts hawthorn residue, 16 parts blueberry pulp, 18 parts Poria cocos powder, and 9 parts xylooligosaccharides.

[0044] Three sets of mulberry and tangerine peel induced extracts were prepared simultaneously, using a mulberry-to-tangerine peel mass ratio of 3:1, an extraction temperature of 55℃, and an extraction time of 3 hours. All other extraction procedures were identical.

[0045] Standard for moisture control in unified layered material laying: The thickness of the bottom layer of the three fermentation beds is uniformly controlled at 6cm to 10cm, the thickness of the middle layer is uniformly controlled at 8cm to 12cm, and the thickness of the top layer of bran is uniformly controlled at 0.5cm to 1cm. The moisture content of the bottom layer material is adjusted to 48%, the moisture content of the middle layer material is adjusted to 32%, and the moisture content of the top layer of bran is adjusted to 20%. The mulberry and tangerine peel induction extract is divided into two portions, which are used for bottom spraying and bottom replenishment during the anaerobic fermentation stage, respectively.

[0046] Unified Standard for Aerobic Pre-fermentation of Bacillus subtilis: In the three groups of experiments, the substrate was sprayed with the same total amount of Bacillus subtilis bacterial solution, and the chamber was pre-fermented at a constant temperature of 34°C for 24 hours. Sterile air was introduced for 10 minutes every 6 hours, and the oxygen volume fraction of the bed was maintained at 8% to 12% throughout the process.

[0047] Unified control standards for compound lactic acid bacteria anaerobic fermentation: After pre-fermentation, spray an equal amount of compound lactic acid bacteria mixed liquid, replace the air inside the chamber with sterile nitrogen three times, and continuously introduce a trace amount of sterile nitrogen to ensure that the oxygen volume fraction of the bed is less than 2.0%. The anaerobic fermentation temperature range is 31℃ to 33℃, and the anaerobic fermentation time is 72h to 96h. Every 24h, lactose solution and remaining induction extract are added to the bottom substrate. The total mass of the material added at one time is 1% to 2% of the dry weight of the bottom substrate.

[0048] Unified post-ripening freeze-drying pulverization and tableting complete process: After all three anaerobic fermentation processes are completed, a 12-hour closed-loop post-ripening conversion is carried out simultaneously. The three layers of materials are mixed and then sent to a vacuum freeze-drying equipment for dehydration treatment with the same freeze-drying parameters. The moisture content of the dried material is controlled to be within 5%. The material is then pulverized and sieved to obtain 90-mesh powder. In the tableting process, the base powder accounts for 96% of the total mass of the chewable tablets, and the internal ratio of excipients remains fixed.

[0049] Summary of performance test results of fermentation products with different substrate ratios: Table 1 shows the detection data of fermentation products with different substrate ratios:

[0050] As shown in Table 1, the test values ​​clearly indicate that the first group had a low proportion of various substrates, resulting in insufficient available carbon sources and functional plant active ingredients during fermentation. This led to the lowest production of short-chain fatty acids, the weakest bacteriocin inhibitory effect, a high water activity value, and poor product storage stability. The third group reached the upper limit of the substrate addition range. The excessive proportion of fruit pomace and Poria cocos reduced the porosity of the fermentation bed, hindering oxygen and nitrogen exchange and reducing anaerobic fermentation efficiency. Its short-chain fatty acid content and inhibitory effect were only better than the first group. The second group had a substrate ratio in the middle range, with a balanced proportion of various raw materials. During fermentation, microorganisms could fully utilize the substrate for metabolism, resulting in the highest accumulation of short-chain fatty acids, the largest bacteriocin inhibition zone diameter, and the lowest water activity value. Overall, its fermentation effect was better than the other two groups. Example 3:

[0051] Optimization and verification of process parameters for the preparation of mulberry and tangerine peel induced extract.

[0052] Experimental setup for gradient ratio of mulberry and dried tangerine peel raw materials: Three different raw material mass ratio gradients were set up, with a uniform extraction temperature of 60℃, a uniform extraction time of 2.5h, and a consistent total amount of purified water added throughout the process.

[0053] The first tier has a mulberry to tangerine peel quality ratio of 3:1.

[0054] The second tier consists of mulberry and dried tangerine peel in a 4:1 ratio.

[0055] The third tier consists of mulberry and dried tangerine peel with a quality ratio of 5:1.

[0056] After the extraction was completed, the solid residue was filtered using a 200-mesh filter cloth, and the clarified induction liquid was collected and used for the addition of exogenous signal substrates in the three-component stratified solid-state fermentation.

[0057] Setting up a comparative experiment on extraction temperature gradient: The ratio of mulberry to dried tangerine peel was kept constant at 4:1, and three sets of constant temperature extraction gradients were set up, with a uniform extraction time of 2.5 hours.

[0058] The first temperature gradient is 55°C.

[0059] The second temperature gradient is 60℃.

[0060] The third temperature gradient is 65℃.

[0061] Setting up a gradient extraction time comparison experiment: The ratio of mulberry to dried tangerine peel was kept constant at 4:1, the extraction temperature was kept constant at 60℃, and three sets of extraction time gradients were set.

[0062] The first duration gradient is 2 hours.

[0063] The second duration gradient is 2.5 hours.

[0064] The third duration gradient is 3 hours.

[0065] The entire fermentation process is implemented in a unified and standardized manner: All gradient groups adopted the substrate ratio obtained from Example 2. The thickness of the three-layer matrix, the material moisture control standard, the aerobic pre-fermentation parameters of Bacillus subtilis, the nitrogen oxygen control conditions for anaerobic fermentation of lactic acid bacteria, and the vacuum freeze-drying, pulverizing and tableting process were all kept consistent. Only the mulberry and tangerine peel induced extracts prepared by different processes in each group were replaced.

[0066] Summary of detection data of fermentation products corresponding to extracts from different processes: Table 2 shows the detection data of fermentation products corresponding to different mulberry and tangerine peel extraction processes:

[0067] As shown in Table 2, observing all the test values, a low mulberry content results in insufficient endogenous signaling substances in the extract, failing to adequately induce the microbial metabolism and synthesis of short-chain fatty acids. Extraction temperatures below 60℃ lead to insufficient dissolution of polyphenols and flavonoids inducing substances from mulberry peel, while temperatures above 60℃ cause decomposition of some heat-sensitive active substances, similarly reducing the induction effect. Extraction times less than 2.5 hours result in incomplete release of active substances from the raw material, while extraction times exceeding 2.5 hours lead to oxidation and loss of some effective components due to prolonged high-temperature soaking. The sample with the 4:1 ratio, 60℃ temperature, and 2.5 hours extraction time shown in the table has the highest short-chain fatty acid value and the lowest water activity; this extraction process is the optimal preparation scheme. Example 4:

[0068] Optimization and verification of parameters such as anaerobic fermentation temperature, fermentation time, and bottom feed addition amount.

[0069] Anaerobic fermentation isothermal temperature gradient grouping experiment: The fermentation substrate ratio, three-layer substrate laying operation, aerobic pre-fermentation parameters of Bacillus subtilis, total amount of lactic acid bacteria inoculation, and nitrogen oxygen control standards were all standardized and fixed. Three constant temperature gradients of 31℃, 32℃, and 33℃ were set for the anaerobic fermentation stage. The anaerobic fermentation time for all groups was standardized and fixed at 84h. Every 24h, the bottom substrate was supplemented with a mixture of lactose solution and mulberry and tangerine peel induction extract.

[0070] Anaerobic fermentation duration gradient grouping experiment: The anaerobic fermentation constant temperature was uniformly set at 32℃, and three anaerobic fermentation time gradients were set at 72h, 84h, and 96h. All supporting processes, including substrate preparation, inoculum addition, feeding operations, freeze-drying and tableting, were standardized and unified throughout the entire process.

[0071] Gradient test of bottom feed addition amount in anaerobic stage: Anaerobic fermentation was carried out at a constant temperature of 32℃ for a fixed fermentation time of 84 hours. Every 24 hours, a mixture of lactose and mulberry and tangerine peel extract was added to the bottom substrate. Three addition gradients were set up: 1% of the dry weight of the bottom substrate, 1.5% of the dry weight of the bottom substrate, and 2% of the dry weight of the bottom substrate.

[0072] Summary of product detection data for different anaerobic process parameters: Table 3 shows the detection data of short-chain fatty acids in products from different anaerobic fermentation processes:

[0073] Table 3 shows that at an anaerobic fermentation temperature of 31℃, the overall metabolic activity of the compound lactic acid bacteria is low, and the synthesis rate of short-chain fatty acids is slow. At 33℃, close to the upper limit of the strain's tolerance, the activity of some Bifidobacteria is slightly inhibited, and the product content decreases. At a fermentation time of 72 hours, the microbial metabolic transformation is incomplete, resulting in insufficient accumulation of effective metabolites. Extending the fermentation time to 96 hours, a small amount of short-chain fatty acids is consumed by the microorganisms under prolonged anaerobic conditions, leading to a slight decrease in product content. A 1% feed addition is insufficient in terms of exogenous carbon source and inducing substances, failing to continuously stimulate the strain's metabolism; a 2% feed addition results in an excessively high substrate concentration, which slightly inhibits bacterial proliferation. In the table, the group with a temperature of 32℃, a fermentation time of 84 hours, and a feed addition of 1.5% (dry weight at the bottom) has the highest short-chain fatty acid values, representing the optimal anaerobic process parameters. Example 5:

[0074] Verification of powder formulation, physicochemical properties, and functional stability of chewable tablets.

[0075] Gradient settings for the ratio of compounded biogenic base powder to food-grade excipients: All fermentation processes were prepared using the optimal process obtained from the previous screening to prepare composite post-biotic base powder, and three sets of tableting powder ratio gradients were set.

[0076] The first ratio is 95 parts base powder and 5 parts food-grade excipients.

[0077] The second ratio is 96.5 parts base powder and 3.5 parts food-grade excipients.

[0078] The third ratio is 98 parts base powder and 2 parts food-grade excipients.

[0079] The internal composition ratio of the three excipient groups is uniform and fixed, with microcrystalline cellulose, sorbitol, and magnesium stearate in a ratio of 22:15:3.

[0080] Unified operating standards for powder mixing and tableting: All three powders were put into a three-dimensional motion mixer and stirred continuously for 30 minutes. The tableting pressure of the rotary tablet press was uniformly kept constant at 20 kN. The theoretical weight of a single tablet after stamping was uniformly 0.8 g. After the tablets were stamped, they were placed in a room temperature and light-proof environment for 72 hours to stabilize the physical properties of the tablets.

[0081] Summary of physicochemical and functional tests of the three groups of finished tablets: Table 4 shows the test data of chewable tablets with different powder ratios:

[0082] Table 4 shows that the first formulation has the highest excipient content, resulting in tablet hardness, slow disintegration in the intestines, delayed release of active ingredients, and a lower improvement in in vitro gut microbiota regulation. Furthermore, the loss of short-chain fatty acids after long-term storage is higher than the second formulation. The third formulation has the lowest excipient content, insufficient powder binding properties, and is extremely prone to breakage during compression, failing to form complete and stable tablets. It also shows the most significant loss of active substances during storage, and its gut microbiota regulation effect is weaker than the second formulation. The second formulation has a moderate amount of excipients, suitable tablet hardness, faster disintegration, better encapsulation and protection of active substances, the highest short-chain fatty acid retention rate after 90 days of storage, and significantly higher increases in Bifidobacteria and Lactobacillus levels in simulated intestinal environments compared to the other two groups. This formulation is the optimal formulation for chewable tablets.

[0083] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A functional food composition for regulating intestinal flora in children, characterized in that, The composition comprises a complete fermentation product obtained by stratified solid-state anaerobic in-situ induced fermentation; The solid substrate used for fermentation includes, by weight, 28-38 parts fermented yam powder, 18-25 parts hawthorn residue, 10-16 parts blueberry pomace, 12-18 parts Poria cocos powder, and 4-9 parts xylooligosaccharides. The fermentation process uses mulberry and tangerine peel induction extract as an exogenous signal substrate. The microbial community used in the fermentation includes Bacillus subtilis, Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei. The microbial community is added in a stepwise inoculation manner. The fermentation system consists of a three-layer matrix structure, which, from bottom to top, consists of a bottom layer of mixed hawthorn residue and blueberry residue, a middle layer of mixed fermented yam powder, poria cocos fragments, and xylooligosaccharides, and a top layer of bran as an isolation layer.

2. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, The fermented yam powder is made from fresh yam through washing, slicing, steaming, drying, and ultra-fine grinding. The thickness of the yam slices is 3mm to 6mm, the steaming time is 25min to 40min, the drying temperature is 50℃ to 55℃, and the moisture content of the dried material is less than 10%. The hawthorn pomace and blueberry pomace are by-products of juice extraction. After being washed and drained, the two types of pomace are crushed to 20-40 mesh. The Poria cocos powder is obtained by pulverizing Poria cocos raw material after washing and drying. The mulberry and tangerine peel induced extract is obtained by mixing mulberry and tangerine peel at a mass ratio of 3:1 to 5:1, adding purified water, and extracting at 55℃ to 65℃ for 2 to 3 hours, followed by filtration to remove solid residue.

3. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, After the three-layer substrate is laid, the moisture content of the bottom layer of hawthorn residue and blueberry residue mixture is adjusted to 48%, and half of the total amount of mulberry and tangerine peel induced extract is sprayed onto the bottom layer. The moisture content of the mixture of medium-fermented yam powder, chopped Poria cocos, and xylooligosaccharides is adjusted to 32%. The top layer of bran separator should be 0.5cm to 1cm thick, and the moisture content of the separator should be adjusted to 20%. The thickness of the bottom layer is 6cm to 10cm, and the thickness of the middle layer is 8cm to 12cm.

4. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, The Bacillus subtilis was inoculated into the middle layer substrate in the aerobic pre-fermentation enzymatic hydrolysis and activation process, with a constant pre-fermentation temperature of 32℃~36℃ and a total duration of 22h~26h. Sterile air was introduced every 6 hours during the pre-fermentation cycle, with a single aeration time of 8 to 12 minutes, and the oxygen volume fraction in the bed was maintained at 8% to 12%. A mixed culture of Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei was inoculated into the middle and bottom substrates after pre-fermentation.

5. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, After the lactic acid bacteria mixed bacterial solution is introduced, the air inside the fermentation bed cavity is replaced three times with sterile nitrogen. A small amount of sterile nitrogen is continuously introduced to maintain a positive pressure anaerobic environment in the cavity, and the oxygen volume fraction in the bed is controlled to be below 2.0%. The anaerobic fermentation temperature range is 31℃~33℃, and the anaerobic fermentation duration is 72h~96h. During the anaerobic fermentation stage, lactose solution and the remaining mulberry and tangerine peel induction extract are added to the bottom substrate every 24h. The total mass of the materials added at one time is 1%~2% of the dry weight of the bottom substrate.

6. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, The water activity of the whole-component fermentation product is not greater than 0.6, and the whole-component fermentation product contains short-chain fatty acids and bacteriocins produced by the fermentation process.

7. The functional food composition for regulating intestinal flora in children according to claim 1, characterized in that, After all fermentation processes are completed, the bottom, middle, and top layers of materials are thoroughly mixed. The mixture is then vacuum freeze-dried to remove free moisture, and the moisture content of the dried material is controlled to be no more than 5%. The dried material is pulverized and sieved to obtain a composite post-genetic matrix powder of 80-100 mesh.

8. The functional food composition for regulating children's intestinal flora according to claim 1, characterized in that, The composite post-biotic base powder is mixed evenly with food-grade excipients and then fed into a rotary tablet press to be pressed into chewable tablets. The base powder accounts for 95% to 98% of the total mass of the chewable tablets, and food-grade excipients account for 2% to 5%. Food-grade excipients include microcrystalline cellulose, sorbitol, and magnesium stearate.

9. A method for preparing a functional food composition for regulating intestinal flora in children, the method being applicable to the functional food composition for regulating intestinal flora in children as described in any one of claims 1-8, characterized in that, The specific steps of this method are as follows: S1, complete the standardized pretreatment of fermented yam powder, hawthorn residue, blueberry residue and poria cocos powder, and simultaneously complete the extraction and solid-liquid separation of mulberry and tangerine peel induced extract, and collect all solid raw materials and liquid induced extract for later use. S2, In the solid fermentation bed chamber, the bottom layer material, the middle layer material and the top layer bran isolation layer are laid from bottom to top in sequence, and the moisture content of each layer of material is adjusted layer by layer to seal the fermentation bed chamber; S3, the Bacillus subtilis bacterial solution that has been activated step by step is evenly sprayed into the middle layer of the substrate, the middle layer material is stirred and mixed, the fermentation bed is sealed and a constant temperature is set, and sterile air is intermittently introduced to maintain a specific oxygen concentration in the chamber for constant temperature aerobic pre-fermentation. S4. After pre-fermentation, stop the introduction of sterile air. Spray the activated Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei mixed bacterial solution onto the middle and bottom substrates and stir the materials. Introduce sterile nitrogen to replace the air inside the chamber. Continuously introduce a small amount of sterile nitrogen to maintain a positive pressure anaerobic environment inside the chamber. Set a constant fermentation temperature for anaerobic fermentation. During the anaerobic fermentation cycle, add lactose solution and the remaining mulberry and tangerine peel induction extract to the bottom substrate at regular intervals. The total mass of each addition is 1% to 2% of the dry weight of the bottom substrate. S5. After the anaerobic induced co-fermentation process is completed, the temperature control parameters of the fermentation bed are lowered to keep the fermentation bed in a closed state and continue to be statically fermented for post-ripening conversion. S6. After the post-ripening conversion is completed, the fermentation bed is turned on, and the bottom, middle and top layers of materials are fully mixed to obtain mixed fermentation material. The mixed fermentation material is sent to a vacuum freeze dryer for dehydration treatment. The dried material is then pulverized and sieved to obtain base powder. The base powder is fully mixed with food-grade excipients, and the mixed powder is sent to a rotary tablet press to be pressed into chewable tablets.

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