High-affinity glutamine intestinal conditioning flora protein powder and preparation method thereof

Abstract

The invention belongs to the technical field of functional food, and particularly relates to high-affinity glutamine intestinal flora regulating protein powder and a preparation method thereof. The invention relates to high-affinity glutamine intestinal flora regulating protein powder, which is prepared from 62 to 68 percent of enzymolysis whey protein peptide, 19 to 21 percent of acetylated glutamine and 11 to 14 percent of compound prebiotics. The preparation method comprises the processes of enzymolysis, modification, after-treatment, compounding and drying, through molecular-level collaborative design, 600 + / -50 Da is adopted for enzymolysis of whey peptide, and gamma-glutamyl glutamine dipeptide is enriched to be larger than or equal to 6%; acetylated glutamine is utilized for targeted modification, the acetylation degree is larger than or equal to 98%, and isopeptide bonds are formed; the exposure rate of beta-glucan is greater than or equal to 82% due to the prebiotics subjected to ice-bath ultrasonication, and the glycopeptide compound is combined, so that the bioavailability of glutamine is greater than or equal to 95%, the bifidobacterium is proliferated by more than 3 times, the blood lactic acid clearance rate is increased by 40%, and the advantages of stable dissolution, fermentation promoting effect, effervescent dosage form and the like are achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of functional foods, and particularly relates to a high-affinity glutamine intestinal flora regulating protein powder and a preparation method thereof. Background Art

[0002] Intestinal health and motor function recovery are core research areas in modern nutrition. As the primary energy source for intestinal epithelial cells, glutamine plays a key role in maintaining intestinal barrier integrity, regulating microbial balance, and repairing tissues after exercise. However, traditional glutamine supplements have significant technical bottlenecks: First, oral bioavailability is low, the acidic environment of the stomach can cause over 40% of the ingredients to degrade, and free glutamine is easily competitively consumed by pathogenic bacteria in the intestines. Second, a single ingredient cannot simultaneously achieve microbial regulation and mucosal repair, and insufficient intestinal colonization efficiency leads to significant efficacy limitations. Third, existing formulations, such as ordinary powder tablets, have problems such as poor stability and slow absorption rates, which cannot meet the demand for rapid onset and portability in the field of sports nutrition.

[0003] In the existing technology, some solutions use large molecular whey protein peptides with a molecular weight greater than 2kDa to be combined with free glutamine. However, the high molecular weight peptides require secondary enzymatic hydrolysis in the intestines before they can be absorbed, resulting in low nutrient delivery efficiency. In addition, the compounding system lacks a targeted protection mechanism, and the degradation rate of glutamine in the stomach is high. Other studies have attempted to introduce prebiotics such as oligofructose to regulate the microbiome, but the acid stability problem of the core ingredients has not been solved, and the synergistic mechanism is unclear. In terms of the application of postbiotics, traditional processes mostly use the method of directly adding inactivated bacteria, resulting in insufficient exposure of cell wall active ingredients such as β-glucan, limited immune regulatory activity and intestinal adhesion ability. In addition, commercially available sports nutrition preparations generally have defects such as poor solubility, reconstitution time greater than 5 minutes, and a single dosage form of only conventional powders, which cannot take into account both storage and transportation stability and terminal user experience.

[0004] From a mechanism of action perspective, due to their large molecular weight, conventional whey protein peptides only physically combine with glutamine, failing to form a stable protective structure. This results in the separate release of both in the gastrointestinal tract, increasing the risk of glutamine being destroyed by gastric acid and hindering the synergistic effect of enhancing intestinal absorption efficiency. While the addition of prebiotics alone can promote the proliferation of beneficial bacteria, the lack of a coordinated design with the glutamine delivery system prevents the spatial and temporal coordination between microbial regulation and mucosal repair. The limitations of postbiotic preparation processes lead to insufficient release of their active ingredients, making it difficult to effectively activate the intestinal immune response, limiting their potential for enhancing barrier function.

[0005] In terms of formulation technology, traditional protein powders, due to their high proportion of hydrophobic amino acids, are prone to forming intermolecular hydrogen bonds during storage, leading to clumping. This requires prolonged stirring for reconstitution, compromising ease of use. Conventional dosage forms such as tablets disintegrate slowly, failing to meet the need for rapid post-exercise nutritional supplementation. Simple granule designs fail to consider compatibility with the intestinal pH environment, potentially leading to premature release or degradation of active ingredients in the stomach.

[0006] In summary, existing technologies have yet to address core issues such as low glutamine bioavailability, insufficient synergy between microbiome regulation and barrier repair, and suboptimal formulation performance. The root cause lies in a lack of multi-layered technological innovation, from molecular structure design and delivery system optimization to the synergy of functional components. Achieving targeted glutamine protection, synergistic regulation of the microbiome-mucosal axis, and rapid nutrient absorption through precise chemical modification, efficient active ingredient release mechanisms, and intelligent dosage form design remains a key technical challenge in this field. Summary of the Invention

[0007] The purpose of the present invention is to provide a high-affinity glutamine intestinal flora regulating protein powder and a preparation method thereof.

[0008] In order to achieve the above object, the present invention provides the following technical solutions:

[0009] A high-affinity glutamine intestinal flora regulating protein powder, comprising, by mass percentage:

[0010] 62-68% enzymatically hydrolyzed whey protein peptides: 90wt% of the peptides have a molecular weight of <1kDa, an average molecular weight of 600±50Da, of which γ-glutamylglutamine dipeptide accounts for ≥6% and hydrophobic amino acids account for ≤12%;

[0011] 19-21% acetylated glutamine: acetylation degree ≥ 98%, residual free glutamine ≤ 0.5%;

[0012] 10-14% compound prebiotics: oligofructose purity ≥95%, resistant dextrin solubility ≥85%, the mass ratio of oligofructose, resistant dextrin and oligogalactose is 1:1.2:0.5, and contains 0.3%-0.5% polydextrose;

[0013] 3-5% inactivated Lactobacillus plantarum postbiotic powder: inactivated at 121°C / 15 min and ultrasonically broken in an 800W ice bath for 10 min, particle size D90 ≤ 1.5 μm, exopolysaccharide ≥ 150 mg / g, β-glucan exposure rate ≥ 80%, and short-chain fatty acid retention rate ≥ 90%.

[0014] Furthermore, the peptide segment containing 4-7 consecutive hydrophilic amino acids in the enzymatically hydrolyzed whey protein peptide accounts for ≥30%, and the complex of the enzymatically hydrolyzed whey protein peptide and acetylated glutamine accounts for ≥88%.

[0015] Furthermore, the galacto-oligosaccharide and polyglucose in the composite prebiotics are subjected to Maillard reaction in the presence of whey protein peptide at 60° C. and 75% relative humidity for 4 hours to generate a glycopeptide complex with a molecular weight of 500-2000 Da.

[0016] A method for preparing a high-affinity glutamine intestinal flora-regulating protein powder, comprising:

[0017] (a) Enzymatic hydrolysis: Whey protein was hydrolyzed with a mixed protease containing 2.5% whey protein and a neutral protease to trypsin ratio of 4:1 at pH 7.0 ± 0.2 and 58°C for 50 min. The target peptides were separated by a 10 kDa ultrafiltration membrane.

[0018] (b) Modification: The enzymatic peptide was mixed with acetylated glutamine at a mass ratio of 3.2:1, and 0.25% transglutaminase was added. The mixture was reacted at 42°C and pH 6.9 for 2.5 h to form an ε-(γ-glutamyl)lysine isopeptide bond.

[0019] (c) Postbiotic treatment: Lactobacillus plantarum cells were inactivated at 121°C for 15 min, ultrasonically disrupted at 800 W for 10 min in an ice bath at ≤10°C, and freeze-dried to obtain a powder.

[0020] (d) Compounding: Mixing the product of step (b), the composite prebiotics, and the postbiotic powders in a nitrogen environment with an oxygen content of ≤0.1% for 15 minutes;

[0021] (e) Drying: spray drying, air inlet temperature 58°C, air outlet temperature 33°C, control the powder moisture content ≤ 3.2%, particle size D50 = 18 ± 2 μm.

[0022] An effervescent granule containing high-affinity glutamine intestinal flora regulating protein powder, comprising: 70-75% protein powder, an effervescent system consisting of citric acid and sodium bicarbonate, the mass ratio of citric acid to sodium bicarbonate being 1:1.3, and a 3% hypromellose coating layer;

[0023] Performance: particle size D50 = 500-800 μm, foaming time in water ≤ 25 seconds, re-dissolved pH 6.0-6.5, water vapor transmission rate ≤ 0.1 g / (m 2 day);

[0024] Process: Dry granulation tableting pressure 10-15kN, single tablet mass 5g, disintegration time ≤1 minute.

[0025] Application of a high-affinity glutamine intestinal flora regulating protein powder in the preparation of post-exercise recovery preparations:

[0026] Compound formula: composed of protein powder, coconut water powder and electrolytes, the mass ratio of the three is 85:10:5; the electrolyte is a mixture of sodium chloride and potassium chloride, wherein the mass ratio of sodium chloride to potassium chloride is 3:1;

[0027] Physical properties: Redissolution time at 40°C ≤ 2 minutes, osmotic pressure 270±20mOsm / kg;

[0028] Dosage: The single dose of the preparation is 20g, which contains 17g of protein powder and should be taken within 30 minutes after exercise.

[0029] Application of a high-affinity glutamine intestinal flora regulating protein powder in the preparation of functional foods for improving leaky gut syndrome:

[0030] Dosage: 25g daily, divided into two doses, for 21 consecutive days;

[0031] Biomarker testing: The patient's serum DAO enzyme activity decreased by ≥45%, fecal calprotectin ≤50 μg / g, and the expression of tight junction protein ZO-1 increased by 2 times;

[0032] Structural characteristics: The β-glucan exposure rate of the postbiotic powder is ≥80%.

[0033] Application of a high-affinity glutamine intestinal flora regulating protein powder as a yogurt fermentation promoter:

[0034] Dosage: The dosage is 4% of the milk base mass, of which the postbiotic powder accounts for 4% of the protein powder mass;

[0035] Fermentation conditions: Fermentation at 42°C for ≤6 hours, the number of viable bacteria in the resulting yogurt ≥1×10 10 CFU / mL, water holding capacity ≥85%;

[0036] Product characteristics: pH of finished yogurt is 4.2-4.5, and the content of flavor substance acetaldehyde is ≥25mg / kg.

[0037] A method for detecting high-affinity glutamine intestinal flora regulating protein powder:

[0038] In vitro bacterial flora culture: The inoculated human fecal flora was cultured under anaerobic conditions at 37°C for 24 hours, and the number of bifidobacteria was determined by plate count method;

[0039] Simulated digestion: The gastrointestinal digestion model was prepared using the USP dissolution method, and the glutamine release was determined by HPLC.

[0040] The beneficial effects of the present invention are:

[0041] 1. Significantly improves glutamine bioavailability. Acetylated glutamine forms a covalent isopeptide bond catalyzed by transaminase. Combined with the carrier effect of small molecule peptides, glutamine bioavailability can reach over 95% in a simulated gastrointestinal digestive environment. In comparison, the bioavailability of free glutamine is less than 50%. This technology effectively circumvents the degradation of glutamine by gastric acid.

[0042] 2. Strengthening intestinal flora colonization and barrier repair. After ultrasonic disruption, the exposure rate of β-glucan in the prebiotics reached over 82%. Synergistically acting with a glycopeptide prebiotic complex (fructooligosaccharides, resistant dextrin, and galacto-oligosaccharides in a ratio of 1:1.2:0.5), the proliferation of bifidobacteria increased by more than 3 times in human fecal flora culture experiments. Clinical data showed that serum DAO enzyme activity decreased by over 45% and ZO-1 protein expression increased by 2.1 times, indicating a significant intestinal barrier repair effect.

[0043] 3. Achieve rapid recovery after exercise. The formulation contains electrolytes (NaCl to KCl in a 3:1 ratio) and coconut water powder, with an osmotic pressure controlled at 270±20mOsm / kg, which is consistent with the isotonic absorption characteristics of the intestinal tract. Experiments have shown that a single 20g dose of the formulation increases blood lactate clearance by 40%, meeting nutritional needs during the 30-minute golden window after exercise and accelerating post-exercise recovery.

[0044] 4. Breaking through the bottleneck of protein powder dissolution stability. The enzymatic peptide contains less than 12% hydrophobic amino acids and over 30% continuous hydrophilic peptide segments, enabling the powder to re-dissolve in 40°C water in under 2 minutes. After spray drying, the powder's angle of repose is ≤30°, and its fluidity is 35% higher than similar products, effectively eliminating agglomeration and significantly improving product dissolution stability.

[0045] 5. Expand the application scenarios of high-activity fermented milk. As a yogurt fermentation promoter, when added at a 4% dosage, the content of postbiotic exopolysaccharides is ≥150mg / g, which can stimulate the proliferation of lactic acid bacteria and shorten the fermentation time to within 6 hours (8 hours for the control group), and the number of viable bacteria in the finished product is ≥1×10 10 CFU / mL, acetaldehyde flavor substance content ≥25mg / kg, broadening the application scenarios of high-activity fermented milk.

[0046] 6. Innovative dosage form ensures the terminal experience. Effervescent granules are coated with hypromellose, with a water vapor transmission rate of ≤0.12g / (m 2 ·day), disintegration time in cold water ≤ 1 minute, and pH controlled at 6.0-6.5, adapted to the weakly alkaline environment of the intestines. The aluminum-plastic packaging design extends the product's shelf life to 24 months, effectively ensuring a good user experience. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1The figure is a bar graph comparing the proliferation folds of bifidobacteria.

[0048] Figure 2 It is a bar graph comparing the decrease rate of serum DAO enzyme activity. DETAILED DESCRIPTION

[0049] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention; it is obvious that the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0050] Example 1: Preparation and performance testing of protein powder

[0051] Raw materials and processes:

[0052] 1. Enzymatic hydrolysis:

[0053] Take 1000g of whey protein and add 25g of composite protease (20g of neutral protease and 5g of trypsin);

[0054] The product was hydrolyzed at pH 7.0 and 58°C for 50 minutes, the enzyme was inactivated in a boiling water bath, and the product was separated by a 10 kDa ultrafiltration membrane to obtain a peptide fragment with a molecular weight of <1 kDa (HPLC determination: average molecular weight 602 Da, γ-glutamylglutamine dipeptide accounted for 6.8%, and hydrophobic amino acids accounted for 10.2%).

[0055] 2. Modification:

[0056] 680 g of enzymatic peptide was mixed with 210 g of acetylated glutamine (acetylation degree 98.5%, free glutamine 0.3%), and 2.2 g of transglutaminase was added;

[0057] The reaction was carried out at 42° C. and pH 6.9 for 2.5 hours to form a complex (HPLC determination showed that the complex accounted for 89.1%, and the peptide segments containing ≥5 consecutive hydrophilic amino acids accounted for 32%).

[0058] 3. Postbiotic processing:

[0059] Lactobacillus plantarum cells were inactivated at 121°C for 15 min and ultrasonically disrupted at 800 W for 10 min in an ice bath (particle size D90 = 1.3 μm);

[0060] The powder was freeze-dried to obtain an exopolysaccharide of 158 mg / g, a β-glucan exposure rate of 82%, and a short-chain fatty acid retention rate of 92%.

[0061] 4. Compounding and drying:

[0062] 890 g of the modified product, 120 g of the composite prebiotic (fructooligosaccharide: resistant dextrin: galacto-oligosaccharide = 40 g: 48 g: 20 g, polydextrose 4 g), and 40 g of the postbiotic powder were mixed for 15 min under nitrogen with an oxygen content of 0.08%;

[0063] Spray drying (inlet air 58°C / outlet air 33°C) gave a powder (water content 2.9%, D50=17 μm).

[0064] Performance Verification:

[0065] Maillard reaction product: The compound prebiotics reacted with whey protein peptides at 60°C and 75% relative humidity for 4 hours to generate a glycopeptide complex with a molecular weight of 1500 Da (confirmed by mass spectrometry).

[0066] In vitro bacterial flora culture: After anaerobic culture of human fecal flora for 24 hours, the number of bifidobacteria increased by 3.2 times (1.5 times in the control group).

[0067] Simulated digestion: USP dissolution method was used to determine the release of glutamine after gastrointestinal digestion, which reached 95% (HPLC detection).

[0068] Example 2: Application of effervescent granules

[0069] Formula and process:

[0070] Take 72g of protein powder from Example 1, 10g of citric acid, 13g of sodium bicarbonate, and 3g of hypromellose;

[0071] Dry granulation (pressure 12kN), tableting (single tablet 5g), and coating with hypromellose ethanol solution.

[0072] performance:

[0073] Particle size D50 = 650 μm, foaming time in water 22 seconds, reconstitution pH 6.2;

[0074] Water vapor transmission rate 0.08g / (m 2 ·day), disintegration time 50 seconds.

[0075] Example 3: Sports recovery preparation

[0076] Formula and usage:

[0077] 85g protein powder, 10g coconut water powder, 5g electrolytes (NaCl:KCl=3.75g:1.25g);

[0078] Take a single 20g dose (containing 17g protein powder) 30 minutes after exercise.

[0079] characteristic:

[0080] The reconstitution time at 40°C is 90 seconds, and the osmotic pressure is 265 mOsm / kg (measured by a freezing point osmometer).

[0081] Example 4: Application for improving leaky gut

[0082] Plan and results:

[0083] Patients with leaky gut syndrome should take 25g daily (in two doses) for 21 consecutive days;

[0084] Serum DAO enzyme activity decreased by 52%, fecal calprotectin was 42 μg / g, and ZO-1 protein expression increased by 2.1 times (Western blot verification).

[0085] Example 5: Yogurt fermentation accelerator

[0086] Process and products:

[0087] 4% of the protein powder of Example 1 (containing 1.6 g / L of postbiotic powder) was added to fresh milk.

[0088] Fermentation at 42°C for 5.5 hours, viable bacteria count 1.2×10 10 CFU / mL, water holding capacity 87%.

[0089] pH 4.3, acetaldehyde content 28 mg / kg (determined by GC-MS).

[0090] Comparative Example 1

[0091] Adjustment: A single neutral protease (no trypsin) was used in the enzymatic hydrolysis stage, the average molecular weight of the peptide segment was 850 Da, the γ-glutamylglutamine dipeptide accounted for 3.1%, and the Maillard reaction step was omitted. The remaining parameters were the same as in Example 1.

[0092] result:

[0093] In in vitro bacterial culture, the growth of Bifidobacterium was only 1.8 times;

[0094] Glutamine release after simulated digestion was 78%;

[0095] The yogurt fermentation time was extended to 7 hours, and the number of viable bacteria was 8×10 9 CFU / mL.

[0096] Comparative Example 2

[0097] Adjustment:

[0098] Ultrasonication power was 400 W, postbiotic particle size D90 was 3.2 μm, β-glucan exposure rate was 45%, spray drying inlet air temperature was 65° C., and powder moisture content was 4.0%. Other parameters were the same as those in Example 1.

[0099] result:

[0100] In patients with leaky gut, serum DAO enzyme activity decreased by only 25%;

[0101] The effervescent granules are reconstituted at pH 5.8 and disintegrate in over 2 minutes.

[0102] Table 1: Performance comparison table

[0103]

[0104] Note: Comparative Example 1 was not tested for leaky gut syndrome, and the data are simulated predicted values.

[0105] Application effect association:

[0106] 1. Postbiotic β-glucan exposure rate >80% (Example 1: 82%) is directly associated with increased ZO-1 protein expression in leaky gut syndrome;

[0107] 2. The molecular weight of the Maillard reaction product (1500Da) is in the range of 500-2000Da, which significantly improves the stability of prebiotics.

[0108] The comparison of bifidobacterium proliferation times is shown in the bar graph. Figure 1 The comparison of serum DAO enzyme activity decrease rate is shown in the bar graph. Figure 2 .

[0109] The above description is merely a preferred embodiment of the present invention; however, the scope of protection of the present invention is not limited thereto. Any person skilled in the art who, within the technical scope disclosed by the present invention, makes equivalent substitutions or modifications based on the technical solutions and improved concepts of the present invention shall be covered by the scope of protection of the present invention.

Claims

1. A high-affinity glutamine intestinal flora regulating protein powder, characterized in that: Contains by mass percentage: 62-68% enzymatically hydrolyzed whey protein peptides: 90wt% of the peptides have a molecular weight of <1kDa, an average molecular weight of 600±50Da, of which γ-glutamylglutamine dipeptide accounts for ≥6% and hydrophobic amino acids account for ≤12%; 19-21% acetylated glutamine: acetylation degree ≥ 98%, residual free glutamine ≤ 0.5%; 10-14% compound prebiotics: oligofructose purity ≥95%, resistant dextrin solubility ≥85%, the mass ratio of oligofructose, resistant dextrin and oligogalactose is 1:1.2:0.5, and contains 0.3%-0.5% polydextrose; 3-5% inactivated Lactobacillus plantarum postbiotic powder: inactivated at 121°C / 15 min and ultrasonically broken in an 800W ice bath for 10 min, particle size D90 ≤ 1.5 μm, exopolysaccharide ≥ 150 mg / g, β-glucan exposure rate ≥ 80%, and short-chain fatty acid retention rate ≥ 90%.

2. The high-affinity glutamine intestinal flora regulating protein powder according to claim 1, characterized in that: The enzymatically hydrolyzed whey protein peptide contains a peptide segment containing 4-7 consecutive hydrophilic amino acids accounting for ≥30%, and the complex of the enzymatically hydrolyzed whey protein peptide and acetylated glutamine accounting for ≥88%.

3. The high-affinity glutamine intestinal flora regulating protein powder according to claim 1, characterized in that: The galacto-oligosaccharide and polyglucose in the composite prebiotics are subjected to Maillard reaction in the presence of whey protein peptides at 60° C. and 75% relative humidity for 4 hours to generate a glycopeptide complex with a molecular weight of 500-2000 Da.

4. A method for preparing the high-affinity glutamine intestinal flora regulating protein powder according to any one of claims 1 to 3, comprising: (a) Enzymatic hydrolysis: Whey protein was hydrolyzed with a mixed protease containing 2.5% whey protein and a neutral protease to trypsin ratio of 4:1 at pH 7.0 ± 0.2 and 58°C for 50 min. The target peptides were separated by a 10 kDa ultrafiltration membrane. (b) Modification: The enzymatic peptide was mixed with acetylated glutamine at a mass ratio of 3.2:1, and 0.25% transglutaminase was added. The mixture was reacted at 42°C and pH 6.9 for 2.5 h to form an ε-(γ-glutamyl)lysine isopeptide bond. (c) Postbiotic treatment: Lactobacillus plantarum cells were inactivated at 121°C for 15 min, ultrasonically disrupted at 800 W for 10 min in an ice bath at ≤10°C, and freeze-dried to obtain a powder. (d) Compounding: Mixing the product of step (b), the composite prebiotics, and the postbiotic powders in a nitrogen environment with an oxygen content of ≤0.1% for 15 minutes; (e) Drying: spray drying, air inlet temperature 58°C, air outlet temperature 33°C, control the powder moisture content ≤ 3.2%, particle size D50 = 18 ± 2 μm.

5. An effervescent granule comprising the high-affinity glutamine intestinal flora modulating protein powder according to any one of claims 1 to 3, characterized in that: composition: 70-75% protein powder, an effervescent system composed of citric acid and sodium bicarbonate, the mass ratio of citric acid to sodium bicarbonate is 1:1.3, and a 3% hypromellose coating layer; Performance: particle size D50 = 500-800 μm, foaming time in water ≤ 25 seconds, re-dissolved pH 6.0-6.5, water vapor transmission rate ≤ 0.1 g / (m 2 day); Process: Dry granulation tableting pressure 10-15kN, single tablet mass 5g, disintegration time ≤1 minute.

6. A use of the high-affinity glutamine intestinal flora modulating protein powder according to any one of claims 1 to 3 in the preparation of a post-exercise recovery preparation, characterized in that: Compound formula: composed of protein powder, coconut water powder and electrolytes, the mass ratio of the three is 85:10:5; the electrolyte is a mixture of sodium chloride and potassium chloride, wherein the mass ratio of sodium chloride to potassium chloride is 3:1; Physical properties: Redissolution time at 40°C ≤ 2 minutes, osmotic pressure 270±20mOsm / kg; Dosage: The single dose of the preparation is 20g, which contains 17g of protein powder and should be taken within 30 minutes after exercise.

7. A use of the high-affinity glutamine intestinal flora modulating protein powder according to any one of claims 1 to 3 in the preparation of a functional food for improving leaky gut syndrome, characterized in that: Dosage: 25g daily, divided into two doses, for 21 consecutive days; Biomarker testing: The patient's serum DAO enzyme activity decreased by ≥45%, fecal calprotectin ≤50 μg / g, and the expression of tight junction protein ZO-1 increased by 2 times; Structural characteristics: The β-glucan exposure rate of the postbiotic powder is ≥80%.

8. Use of the high-affinity glutamine intestinal flora regulating protein powder according to any one of claims 1 to 3 as a yogurt fermentation promoter, characterized in that: Dosage: The dosage is 4% of the milk base mass, of which the postbiotic powder accounts for 4% of the protein powder mass; Fermentation conditions: Fermentation at 42°C for ≤6 hours, the number of viable bacteria in the resulting yogurt ≥1×10 10 CFU / mL, water holding capacity ≥85%; Product characteristics: pH of finished yogurt is 4.2-4.5, and the content of flavor substance acetaldehyde is ≥25mg / kg.

9. A method for detecting the high-affinity glutamine intestinal flora modulating protein powder according to any one of claims 1 to 3, characterized in that: In vitro bacterial flora culture: The inoculated human fecal flora was cultured under anaerobic conditions at 37°C for 24 hours, and the number of bifidobacteria was determined by plate count method; Simulated digestion: The gastrointestinal digestion model was prepared using the USP dissolution method, and the glutamine release was determined by HPLC.

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