Probiotic compound beverage with weight-losing function and preparation method of probiotic compound beverage

Abstract

The invention discloses a probiotic composite beverage with a weight losing function and a preparation method thereof, and belongs to the technical field of probiotic beverages, the method comprises the following steps: raw material pretreatment and modification, fermentation system construction, fermentation, filtration and concentration, synergistic auxiliary material preparation, prebiotic solution preparation, microbial inoculum preparation and mixing. Through cooperation of various probiotics, on one hand, intestinal flora balance is adjusted, harmful bacterium growth is inhibited, the number of beneficial bacteria is increased, the intestinal micro-ecological environment is optimized, chronic inflammatory response caused by endotoxin entering blood is reduced, and the obesity risk related to inflammation is reduced; on the other hand, by influencing fat metabolism and sugar metabolism, fat accumulation is reduced, the blood sugar level is reduced, energy consumption is increased, intestinal tracts are stimulated to generate satiety signals, appetite is reduced, caloric intake is reduced, short-chain fatty acid can be generated, fat metabolism is influenced, and therefore the weight losing effect is achieved.

Description

Technical Field

[0001] This invention relates to the field of probiotic beverage technology, specifically to a probiotic compound beverage with weight loss function and its preparation method. Background Technology

[0002] With changing modern lifestyles, obesity has become an increasingly serious global public health challenge. Obesity not only affects an individual's appearance and quality of life, but is also closely related to the development of various chronic diseases, such as cardiovascular disease, diabetes, and hypertension, placing a heavy burden on individual health and the social healthcare system. According to relevant statistics, the proportion of obese people worldwide has been rising continuously in recent years, with some developed countries and regions exceeding 50% of the population. In China, the number of obese people is also growing rapidly, making effective prevention and control of obesity an urgent priority.

[0003] Studies have shown that the gut microbiota plays a crucial role in human health and energy metabolism, and gut microbiota dysbiosis is closely related to obesity. Under normal circumstances, beneficial and harmful bacteria in the gut maintain a dynamic balance, working together to maintain normal gut function. However, when the gut microbiota is imbalanced, harmful bacteria overgrow, producing large amounts of endotoxins. These endotoxins enter the bloodstream, triggering chronic inflammatory responses, which in turn affect fat and glucose metabolism, leading to fat accumulation, elevated blood sugar, and an increased risk of obesity. Simultaneously, the gut microbiota can also influence the appetite center and control total food intake by regulating signal transmission between the gut and the brain. Therefore, regulating the balance of the gut microbiota is considered an important target for preventing and improving obesity.

[0004] Probiotics, as a class of live microorganisms beneficial to the host, have received widespread attention in the field of weight management in recent years. Numerous studies have confirmed that probiotics can regulate the balance of gut microbiota through multiple mechanisms, inhibiting the growth of harmful bacteria, increasing the number of beneficial bacteria, and optimizing the gut microecological environment. On the one hand, probiotics can reduce the chronic inflammatory response caused by endotoxins entering the bloodstream, lowering the risk of inflammation-related obesity; on the other hand, by affecting fat and glucose metabolism, they reduce fat accumulation, lower blood sugar levels, increase energy expenditure, and stimulate the gut to produce satiety signals, reducing appetite and calorie intake. Furthermore, probiotics can produce beneficial metabolites such as short-chain fatty acids during fermentation, which further affect fat metabolism, thereby achieving weight loss. However, current probiotic products on the market still have many shortcomings in terms of strain selection, formulation, and efficacy, resulting in inconsistent weight loss effects.

[0005] Based on this, the present invention designs a probiotic compound beverage with weight loss function and its preparation method to solve the above problems. Summary of the Invention

[0006] To address the aforementioned shortcomings of existing technologies, this invention provides a method for preparing a probiotic compound beverage with weight-loss function, comprising the following steps: S1. Raw material pretreatment and modification; Soak the green plums in an aqueous solution of chlorine dioxide, rinse them with clean water, dry them, and then crush them. S2. Construct the fermentation system; Kluyveromycin was cultured at 28-32℃ for 12-18 hours. Lactobacillus helveticus was cultured at 30-35℃ for 18-24 hours; Acetobacter pasteurellium was cultured at 32-35℃ for 24-36 hours; The fermentation system was obtained by mixing Kluyveromyces martensii, Lactobacillus helveticus, and Acetobacter pasteurella multocida in a volume ratio of 3-5:2-4:1-3. S3. Fermentation; The crushed plum powder is mixed with syrup and introduced into the fermentation system. In the early stage of fermentation, the temperature is controlled at 25-28℃ for anaerobic fermentation for 2-3 days. The temperature is then raised to 30-33℃, and air containing carbon dioxide is introduced for micro-aerobic fermentation for 3-5 days. Ultrasonic fermentation is used to assist fermentation throughout the entire process. S4. Filtration and concentration; After fermentation, ultrafiltration membranes are used to remove microbial cells and macromolecular impurities from the fermentation broth to obtain a clarified fermentation broth. The fermentation broth is then vacuum concentrated to 1 / 5 to 1 / 8 of its original volume and freeze-dried at -30°C to obtain plum fermentation powder. S5. Preparation of synergistic excipients; Add the green plum baking powder to the resistant dextrin and continue stirring to obtain the synergistic excipient; S6. Prepare a prebiotic solution; Weigh out 3-5 parts of inulin, 2-4 parts of fructooligosaccharides, and 1-2 parts of xylooligosaccharides, add them to 100 parts of warm water at 40-45℃, stir well and let them dissolve completely to obtain a prebiotic solution. S7. Preparation of microbial agents; Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were each inoculated into the culture medium and cultured to the logarithmic growth phase. The culture temperature for Lactobacillus gasseri is 30-37℃, and the pH is 6.0-7.0. The culture temperature for Lactobacillus acidophilus is 35-38℃, and the pH is 5.5-6.0. The culture temperature for Lactobacillus johnsonii and Lactobacillus plantarum is 30-35℃, and the pH is 6.5-7.5. The culture temperature for Bifidobacterium adolescentis is 36-38℃, and the pH is 6.5-7.2. The culture temperature for *Lactobacillus paracasei* is 37-40℃, and the pH is 6.8-7.5. The cultured bacterial solution was centrifuged to separate the bacterial cells, which were then washed 2-3 times with sterile water. The bacterial cells were then freeze-dried to obtain freeze-dried bacterial powder of each strain. Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were mixed evenly in a mass ratio of 1-3:2-4:2-4:3-5:2-5:2-3 to obtain the bacterial agent; The total number of viable bacteria in the bacterial agent is greater than or equal to 10. 9 CFU / g; S8. Mixing; Add 2-4 parts of bacterial agent, 8-11 parts of prebiotic solution, and 1-3 parts of synergistic excipients to a mixing tank, stir well and dry to obtain a probiotic compound drink with weight loss function.

[0007] Furthermore, S1 specifically involves: soaking the washed green plums in an aqueous solution containing 0.1-0.3% chlorine dioxide for 15-25 minutes, rinsing them thoroughly with clean water, and then drying the green plums using vacuum freeze-drying technology. Under a vacuum of 0.01-0.03 MPa and a temperature of -30℃ to -40℃, the moisture content of the green plums is reduced to 10-15%. Finally, the dried green plums are pulverized into ultrafine powder with a particle size of 5-10 μm using an air jet mill.

[0008] Furthermore, in S2, the culture medium used to cultivate Kluyveromyces martensii consists of 30-60 g / L glucose, 10-30 g / L lactose, 12-25 g / L yeast extract, 5-15 g / L peptone, 2-5 g / L KH2PO4, 0.5-1.5 g / L MgSO4·7H2O, 1-3 g / L NaCl, 10-50 μg / L vitamin B7, 5-20 mg / L vitamin B3, 5-15 g / L mannitol, and 1-5 g / L wheat peptides.

[0009] Furthermore, in S2, the culture medium used to cultivate Lactobacillus helveticus consists of 20-40 g / L lactose, 10-20 g / L glucose, 15-25 g / L yeast extract, 5-10 g / L yeast peptone, 0.05-0.08 mol / L sodium citrate, 0.5-1.5 g / L MgSO4·7H2O, 0.05-0.15 g / L MgSO4·H2O, 5-20 mg / L vitamin B3, 5-15 g / L mannitol, and 0.5-2 g / L L-cysteine ​​hydrochloride.

[0010] Furthermore, in S2, the culture medium used to cultivate *Acetobacter pasteurellis* consists of 30-45 g / L anhydrous ethanol, 5-15 g / L glucose, 20-35 g / L yeast extract, 5-12 g / L corn steep liquor powder, 3-8 g / L sodium citrate, 1-3 g / L betaine, 0.8-1.5 g / L MgSO4·7H2O, 2-4 g / L KH2PO4, 0.05-0.15 g / L MnSO4·H2O, 10-30 mg / L vitamin B5, 5-15 mg / L para-aminobenzoic acid, 0.5-2 g / L wheat oligopeptide, and 1-3 g / L trehalose.

[0011] Furthermore, S3 specifically involves mixing the pulverized plum powder with a high-concentration syrup of 30-40% by mass, with a syrup-to-plum powder mass ratio of 3-5:1. This mixture is then introduced into the fermentation system, with the inoculation amount accounting for 5-10% of the total volume of the fermentation system. In the initial stage of fermentation, the temperature is controlled at 25-28℃ for anaerobic fermentation for 2-3 days. The temperature is then raised to 30-33℃, and filtered, sterilized air containing 5-10% carbon dioxide is introduced for micro-aerobic fermentation for 3-5 days. Throughout the fermentation process, ultrasonic waves are used to assist fermentation, with ultrasonic treatment performed every 4-6 hours. The ultrasonic frequency is 20-40kHz, the power is 100-200W, and the treatment time is 15-25 minutes.

[0012] Furthermore, S5 specifically involves: adding plum fermentation powder into a mixing tank to obtain a mixed system; first, setting the stirring speed to 60-80 r / min; heating to 55-60℃ at a rate of 1-3℃ / min; continuing to stir at this temperature for 30-35 min; then cooling to 40-45℃ at a rate of 2-4℃ / min; adding 4-6.5% resistant dextrin by weight of the plum fermentation powder; and continuing to stir for 25-30 min to obtain the synergistic excipient.

[0013] Furthermore, in S7, the culture medium used to cultivate Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei consists of 8-12 parts peptone, 6-10 parts beef extract, 3-5 parts yeast extract, 18-22 parts glucose, 0.1-0.3 parts magnesium sulfate, 4-6 parts sodium acetate, 1-3 parts diammonium citrate, 1-3 parts dipotassium hydrogen phosphate, 0.03-0.05 parts manganese sulfate, 0.8-1.2 parts Tween 80, and 1000 parts water.

[0014] A probiotic compound beverage with weight loss function prepared according to the preparation method of the probiotic compound beverage with weight loss function.

[0015] Compared with the prior art, the beneficial effects of this invention are as follows: 1. This invention utilizes the synergistic effect of multiple probiotics to regulate the balance of intestinal flora, inhibit the growth of harmful bacteria, increase the number of beneficial bacteria, optimize the intestinal microecological environment, reduce chronic inflammatory responses caused by endotoxins entering the bloodstream, and lower the risk of inflammation-related obesity. On the other hand, by affecting fat and sugar metabolism, it reduces fat accumulation, lowers blood sugar levels, increases energy consumption, stimulates the intestine to produce satiety signals, reduces appetite, lowers calorie intake, and can also produce short-chain fatty acids, affecting fat metabolism, thereby achieving weight loss effects.

[0016] 2. This invention utilizes highly efficient extraction of plant proteases, lipases, and other active enzymes, as well as dietary fiber, from fermented plum powder to help break down proteins and fats in food, improve digestion and absorption efficiency, and reduce fat accumulation. The dietary fiber it contains can, on the one hand, increase intestinal peristalsis, soften stool to improve constipation, and accelerate the excretion of metabolic waste; on the other hand, dietary fiber has a strong adsorption capacity, which can adsorb intestinal toxins, optimize the intestinal environment, and reduce fat absorption. Furthermore, fermented plum powder can regulate signal transmission between the intestine and the brain, affecting the appetite center, reducing appetite, and decreasing total food intake (the lower food intake in rats in Comparative Examples 1 and 3 can help verify this), thus having a positive effect on weight loss.

[0017] 3. This invention enhances the colonization and metabolic functions of probiotics through a reasonable ratio of bacterial strains, allowing multiple probiotics to work synergistically, effectively regulating the balance of intestinal flora, secreting more beneficial metabolites such as short-chain fatty acids, thereby better inhibiting fat absorption and promoting fat decomposition, achieving a good weight loss effect.

[0018] 4. The enzymes produced during the fermentation process can promote the release and transformation of active components related to fat metabolism in green plums, such as polyphenols, enhancing their ability to promote fat decomposition. On the other hand, the green plum fermentation powder provides a richer and more unique nutritional environment for the microorganisms, which is conducive to better colonization and reproduction, strengthening their functions in regulating intestinal flora balance and inhibiting fat absorption. Simultaneously, both factors work together to optimize the intestinal microecology, stimulating the intestines to produce more beneficial metabolites such as short-chain fatty acids. These substances can affect fat metabolism and regulate appetite signals, exerting a multi-faceted effect in weight loss. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0020] Example 1: This example provides a method for preparing a probiotic compound beverage with weight loss function, including the following steps: S1. Raw material pretreatment and modification; Select high-quality, mature, and pest-free green plums. Rinse the surface with running water to remove impurities. Soak the green plums in a 0.3% chlorine dioxide solution for 25 minutes to effectively sterilize and reduce microbial contamination. Rinse thoroughly with clean water. Use vacuum freeze-drying technology to dry the green plums at a vacuum of 0.03 MPa and a temperature of -40℃, reducing the moisture content of the green plums to 15% (this step not only quickly removes moisture but also retains the nutritional components and flavor substances of the green plums to the greatest extent). Use an air jet mill to pulverize the dried green plums into ultrafine powder with a particle size of 10μm (increasing the contact area between the green plums and fermentation bacteria and other components, significantly improving fermentation efficiency and effect). S2. Construct the fermentation system; Kluyveromyces martensii (CICC 1911) was cultured at 32℃ for 18 h (culture medium composition: 60 g / L glucose, 30 g / L lactose, 25 g / L yeast extract, 15 g / L peptone, 5 g / L KH2PO4, 1.5 g / L MgSO4·7H2O, 3 g / L NaCl, 50 μg / L vitamin B7, 20 mg / L vitamin B3, 15 g / L mannitol, 5 g / L wheat peptides). Lactobacillus helveticus (R0052) was cultured at 35℃ for 24 h (culture medium composition: 40 g / L lactose, 20 g / L glucose, 25 g / L yeast extract, 10 g / L yeast peptone, 0.08 mol / L sodium citrate, 1.5 g / L MgSO4·7H2O, 0.15 g / L MgSO4·H2O, 20 mg / L vitamin B3, 15 g / L mannitol, 2 g / L L-cysteine ​​hydrochloride). Acetobacter pasteurellium (CICC 20769) was cultured at 35℃ for 36 h (culture medium composition: 45 g / L anhydrous ethanol, 15 g / L glucose, 35 g / L yeast extract, 12 g / L corn steep liquor powder, 8 g / L sodium citrate, 3 g / L betaine, 1.5 g / L MgSO4·7H2O, 4 g / L KH2PO4, 0.15 g / L MnSO4·H2O, 30 mg / L vitamin B5, 15 mg / L para-aminobenzoic acid, 2 g / L wheat oligopeptide, 3 g / L trehalose). The fermentation system was obtained by mixing Kluyveromyces martensii, Lactobacillus helveticus, and Acetobacter pasteurella multocida in a volume ratio of 5:4:3. S3. Fermentation; The ultra-finely pulverized plum powder was mixed with a high-concentration syrup (made from white sugar and honey in a 2:1 mass ratio) of 40% by mass, with a syrup-to-plum powder mass ratio of 5:1. This mixture was then introduced into the fermentation system, with the inoculation amount accounting for 10% of the total volume of the fermentation system. In the early stage of fermentation, the temperature was controlled at 28℃ for anaerobic fermentation for 3 days. The temperature was then raised to 33℃, and filtered and sterilized air containing 10% carbon dioxide was introduced for micro-aerobic fermentation for 5 days. Throughout the fermentation process, ultrasonic waves were used to assist fermentation. Ultrasonic treatment was performed every 6 hours, with an ultrasonic frequency of 40kHz, a power of 200W, and a treatment time of 25 minutes. S4. Filtration and concentration; After fermentation, the microbial cells and macromolecular impurities in the fermentation broth were removed using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da to obtain a clarified fermentation broth. The fermentation broth was then vacuum concentrated to 1 / 8 of its original volume and freeze-dried at -30℃ to obtain plum fermentation powder. S5. Preparation of synergistic excipients; The green plum fermentation powder was added to a mixing tank to obtain a mixed system. The stirring speed was first set to 80 r / min, and the temperature was increased to 60℃ at a rate of 3℃ / min. The mixture was stirred at this temperature for 35 min. The temperature was then decreased to 45℃ at a rate of 4℃ / min. 6.5% of the green plum fermentation powder mass of resistant dextrin was added, and the mixture was stirred for another 30 min. During this process, the resistant dextrin combined with the active ingredients in the liquid through intermolecular hydrogen bonds to form a stable colloidal dispersion system, thus obtaining the synergistic excipient. S6. Prepare a prebiotic solution; Weigh out 5 parts inulin, 4 parts fructooligosaccharide, and 2 parts xylooligosaccharide, add them to 100 parts of 45℃ warm water, stir well and let them dissolve completely to obtain a prebiotic solution. S7. Preparation of microbial agents; Lactobacillus gasseri (BNR17), Lactobacillus acidophilus (NCFM), Lactobacillus johnsonii (CICC 10861), Lactobacillus plantarum (LP104), Bifidobacterium adolescentis (CICC 6178), and Lactobacillus paracasei (G15) were each inoculated into the culture medium and cultured to the logarithmic growth phase. The culture temperature for Lactobacillus gasseri was 37℃ and the pH was 7.0. The culture temperature for Lactobacillus acidophilus was 38℃, and the pH was 6.0. Lactobacillus johnsonii and Lactobacillus plantarum were cultured at 35°C and pH 7.5. The culture temperature for Bifidobacterium adolescentis was 38℃, and the pH was 7.2. The culture temperature for *Lactobacillus paracasei* was 40℃, and the pH was 7.5. The culture medium consists of 12 parts peptone, 10 parts beef extract powder, 5 parts yeast powder, 22 parts glucose, 0.3 parts magnesium sulfate, 4-6 parts sodium acetate (for pH adjustment), 3 parts diammonium citrate, 3 parts dipotassium hydrogen phosphate, 0.05 parts manganese sulfate, 1.2 parts Tween 80, and 1000 parts water. The cultured bacterial solution was centrifuged to separate the bacterial cells, which were then washed three times with sterile water. The bacterial cells were then freeze-dried to obtain freeze-dried bacterial powder of each strain. Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were mixed evenly in a mass ratio of 3:4:4:5:5:3 to obtain the bacterial agent; The total number of viable bacteria in the bacterial agent is greater than or equal to 10. 9 CFU / g; S8. Mixing; Add 4 parts of bacterial agent, 11 parts of prebiotic solution, and 3 parts of synergistic excipients to a mixing tank, stir well, and dry at room temperature to obtain a probiotic compound drink with weight loss function.

[0021] Example 2: This example provides a method for preparing a probiotic compound beverage with weight loss function, including the following steps: S1. Raw material pretreatment and modification; Select mature, disease-free, high-quality green plums, rinse off surface impurities with running water, soak the green plums in an aqueous solution containing 0.1% chlorine dioxide for 15 minutes to effectively sterilize and reduce microbial contamination, rinse them clean with water, and dry the green plums using vacuum freeze-drying technology. Under a vacuum of 0.01 MPa and a temperature of -30℃, the moisture content of the green plums is reduced to 10%. The dried green plums are then pulverized into ultrafine powder with a particle size of 5μm using an airflow pulverizer. S2. Construct the fermentation system; Kluyveromyces martensii was cultured at 28°C for 12 h (culture medium composition: 30 g / L glucose, 10 g / L lactose, 12 g / L yeast extract, 5 g / L peptone, 2 g / L KH2PO4, 0.5 g / L MgSO4·7H2O, 1 g / L NaCl, 10 μg / L vitamin B7, 5 mg / L vitamin B3, 5 g / L mannitol, 1 g / L wheat peptide). Lactobacillus helveticus was cultured at 30℃ for 18 hours (culture medium composition: 20 g / L lactose, 10 g / L glucose, 15 g / L yeast extract, 5 g / L yeast peptone, 0.05 mol / L sodium citrate, 0.5 g / L MgSO4·7H2O, 0.05 g / L MnSO4·H2O, 5 mg / L vitamin B3, 5 g / L mannitol, 0.5 g / L L-cysteine ​​hydrochloride). Acetobacter pasteurellum was cultured at 32℃ for 24 h (culture medium composition: 30 g / L anhydrous ethanol, 5 g / L glucose, 20 g / L yeast extract, 5 g / L corn steep liquor powder, 3 g / L sodium citrate, 1 g / L betaine, 0.8 g / L MgSO4·7H2O, 2 g / L KH2PO4, 0.05 g / L MnSO4·H2O, 10 mg / L vitamin B5, 5 mg / L para-aminobenzoic acid, 0.5 g / L wheat oligopeptide, 1 g / L trehalose). The fermentation system was obtained by mixing Kluyveromyces martensii, Lactobacillus helveticus, and Acetobacter pasteurella multocida in a volume ratio of 3:2:1. S3. Fermentation; The ultra-finely pulverized plum powder was mixed with a high-concentration syrup (made from white sugar and honey in a 2:1 mass ratio) of 30% by mass, with the syrup to plum powder mass ratio being 3:1. This mixture was then introduced into the fermentation system, with the inoculation amount accounting for 5% of the total volume of the fermentation system. In the early stage of fermentation, the temperature was controlled at 25℃ for anaerobic fermentation for 2 days. The temperature was then raised to 30℃, and filtered and sterilized air containing 5% carbon dioxide was introduced for micro-aerobic fermentation for 3 days. Throughout the fermentation process, ultrasonic waves were used to assist fermentation. Ultrasonic treatment was performed every 4 hours, with an ultrasonic frequency of 20kHz, a power of 100W, and a treatment time of 15 minutes. S4. Filtration and concentration; After fermentation, microbial cells and macromolecular impurities in the fermentation broth were removed using an ultrafiltration membrane with a molecular weight cutoff of 1000 Da to obtain a clarified fermentation broth. The fermentation broth was then vacuum concentrated to 1 / 5 of its original volume and freeze-dried at -30℃ to obtain plum fermentation powder. S5. Preparation of synergistic excipients; The plum fermentation powder was added to a mixing tank to obtain a mixed system. The stirring speed was set to 60 r / min, and the temperature was increased to 55℃ at a rate of 1℃ / min. The mixture was stirred at this temperature for 30 min. The temperature was then decreased to 40℃ at a rate of 2℃ / min. 4% of the plum fermentation powder mass of resistant dextrin was added, and the mixture was stirred for another 25 min. During this process, the resistant dextrin combined with the active ingredients in the liquid through intermolecular hydrogen bonds to form a stable colloidal dispersion system, thus obtaining the synergistic excipient. S6. Prepare a prebiotic solution; Weigh out 3 parts inulin, 2 parts fructooligosaccharide, and 1 part xylooligosaccharide, add them to 100 parts of 40℃ warm water, stir well and let them dissolve completely to obtain a prebiotic solution. S7. Preparation of microbial agents; Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were each inoculated into the culture medium and cultured to the logarithmic growth phase. The culture temperature for Lactobacillus gasseri was 30℃ and the pH was 6.0. The culture temperature for Lactobacillus acidophilus was 35℃ and the pH was 5.5. Lactobacillus johnsonii and Lactobacillus plantarum were cultured at 30°C and pH 6.5. The culture temperature for Bifidobacterium adolescentis was 36℃, and the pH was 6.5. The culture temperature for *Lactobacillus paracasei* was 37°C, and the pH was 6.8. The culture medium consists of 8 parts peptone, 6 parts beef extract powder, 3 parts yeast powder, 18 parts glucose, 0.1 parts magnesium sulfate, 4-6 parts sodium acetate, 1 part diammonium citrate, 1 part dipotassium hydrogen phosphate, 0.03 parts manganese sulfate, 0.8 parts Tween 80, and 1000 parts water. The cultured bacterial solution was centrifuged to separate the bacterial cells, which were then washed twice with sterile water. The bacterial cells were then freeze-dried to obtain freeze-dried bacterial powder of each strain. Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were mixed evenly in a mass ratio of 1:2:2:3:2:2 to obtain the bacterial agent; The total number of viable bacteria in the bacterial agent is greater than or equal to 10. 9 CFU / g; S8. Mixing; Add 2 parts of bacterial agent, 8 parts of prebiotic solution, and 1 part of synergistic excipient to a mixing tank, stir well, and dry at room temperature to obtain a probiotic compound drink with weight loss function.

[0022] Example 3: This example provides a method for preparing a probiotic compound beverage with weight loss function, including the following steps: S1. Raw material pretreatment and modification; Select mature, disease-free, high-quality green plums, rinse off surface impurities with running water, soak the green plums in an aqueous solution containing 0.2% chlorine dioxide for 22 minutes to effectively sterilize and reduce microbial contamination, rinse them clean with water, and dry the green plums using vacuum freeze-drying technology. Under a vacuum of 0.02 MPa and a temperature of -36℃, the moisture content of the green plums is reduced to 12%. The dried green plums are then pulverized into ultrafine powder with a particle size of 8μm using an airflow pulverizer. S2. Construct the fermentation system; Kluyveromyces martensii was cultured at 31°C for 16 h (culture medium composition: 50 g / L glucose, 22 g / L lactose, 18 g / L yeast extract, 8 g / L peptone, 3 g / L KH2PO4, 1.2 g / L MgSO4·7H2O, 2 g / L NaCl, 40 μg / L vitamin B7, 14 mg / L vitamin B3, 13 g / L mannitol, 4 g / L wheat peptides). Lactobacillus helveticus was cultured at 32℃ for 22 hours (culture medium composition: 34 g / L lactose, 14 g / L glucose, 19 g / L yeast extract, 9 g / L yeast peptone, 0.07 mol / L sodium citrate, 1.1 g / L MgSO4·7H2O, 0.14 g / L MgSO4·H2O, 12 mg / L vitamin B3, 6 g / L mannitol, 1.3 g / L L-cysteine ​​hydrochloride). Acetobacter pasteurellum was cultured at 33℃ for 28 h (culture medium composition: 41 g / L anhydrous ethanol, 12 g / L glucose, 32 g / L yeast extract, 9 g / L corn steep liquor powder, 6 g / L sodium citrate, 2 g / L betaine, 1.2 g / L MgSO4·7H2O, 3 g / L KH2PO4, 0.14 g / L MnSO4·H2O, 25 mg / L vitamin B5, 6 mg / L para-aminobenzoic acid, 1.6 g / L wheat oligopeptide, 2 g / L trehalose). The fermentation system was obtained by mixing Kluyveromyces martensii, Lactobacillus helveticus, and Acetobacter pasteurella in a volume ratio of 4:3:3. S3. Fermentation; The ultra-finely pulverized plum powder was mixed with a high-concentration syrup (made from white sugar and honey in a 2:1 mass ratio) with a mass ratio of 36% to 4:1. The mixture was then introduced into the fermentation system, with the inoculation amount accounting for 8% of the total volume of the fermentation system. In the early stage of fermentation, the temperature was controlled at 27℃ for anaerobic fermentation for 3 days. The temperature was then raised to 32℃, and filtered and sterilized air containing 6% carbon dioxide was introduced for micro-aerobic fermentation for 4 days. Throughout the fermentation process, ultrasonic-assisted fermentation was used. Ultrasonic treatment was performed every 5 hours, with an ultrasonic frequency of 36kHz, a power of 180W, and a treatment time of 21 minutes. S4. Filtration and concentration; After fermentation, the microbial cells and macromolecular impurities in the fermentation broth were removed using an ultrafiltration membrane with a molecular weight cutoff of 2000 Da to obtain a clarified fermentation broth. The fermentation broth was then vacuum concentrated to 1 / 6 of its original volume and freeze-dried at -30℃ to obtain plum fermentation powder. S5. Preparation of synergistic excipients; The green plum fermentation powder was added to a mixing tank to obtain a mixed system. The stirring speed was first set to 70 r / min, and the temperature was increased to 58℃ at a rate of 2℃ / min. The mixture was stirred at this temperature for 32 min. The temperature was then decreased to 43℃ at a rate of 3℃ / min. 5% of the mass of the green plum fermentation powder was added, and the mixture was stirred for another 28 min. During this process, the resistant dextrin combined with the active ingredients in the liquid through intermolecular hydrogen bonds to form a stable colloidal dispersion system, thus obtaining the synergistic excipient. S6. Prepare a prebiotic solution; Weigh out 4 parts inulin, 3 parts fructooligosaccharide, and 2 parts xylooligosaccharide, add them to 100 parts of 43℃ warm water, stir well and let them dissolve completely to obtain a prebiotic solution. S7. Preparation of microbial agents; Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were each inoculated into the culture medium and cultured to the logarithmic growth phase. The culture temperature for Lactobacillus gasseri was 33℃ and the pH was 6.4. The culture temperature for Lactobacillus acidophilus was 36℃ and the pH was 5.8. Lactobacillus johnsonii and Lactobacillus plantarum were cultured at 33°C and pH 6.8. The culture temperature for Bifidobacterium adolescentis was 37℃, and the pH was 7.0. The culture temperature for *Lactobacillus paracasei* was 38°C, and the pH was 7.1. The culture medium consists of 11 parts peptone, 7 parts beef extract powder, 4 parts yeast powder, 21 parts glucose, 0.2 parts magnesium sulfate, 4-6 parts sodium acetate, 2 parts diammonium citrate, 2 parts dipotassium hydrogen phosphate, 0.04 parts manganese sulfate, 0.9 parts Tween 80, and 1000 parts water. The cultured bacterial solution was centrifuged to separate the bacterial cells, which were then washed three times with sterile water. The bacterial cells were then freeze-dried to obtain freeze-dried bacterial powder of each strain. Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were mixed evenly in a mass ratio of 2:3:4:4:3:2 to obtain the bacterial agent; The total number of viable bacteria in the bacterial agent is greater than 10. 10 CFU / g; S8. Mixing; Add 3 parts of bacterial agent, 10 parts of prebiotic solution, and 2 parts of synergistic excipients to a mixing tank, stir well, and dry at room temperature to obtain a probiotic compound drink with weight loss function.

[0023] Comparative Example 1: The difference from Example 3 is that no plum fermentation powder was added in the preparation of the synergistic excipients in this comparative example.

[0024] Comparative Example 2: The difference from Example 3 is that the mass ratio of Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei in this comparative example is 6:3:3:1:7:8.

[0025] Comparative Example 3: The difference from Example 3 is that the mass ratio of Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei in this comparative example is 6:3:3:1:7:8; and no plum fermentation powder was added in the preparation of the synergistic excipient.

[0026] Experimental example: 1. Establishment of a rat obesity model; Strain: SD rats (half male and half female); Age in weeks: 5 weeks old; Starting weight: 180-220g; Number per group: 12; Adaptation period: Feed with standard feed (10% fat, 70% carbohydrates, 20% protein) for 1 week at room temperature of 22±2℃ and 12h light cycle; High-fat feeding period: The diet consisted of 8 weeks of high-fat diet, with 60% fat (lard:soybean oil = 7:3), 17.5% protein, 20% carbohydrates, 2% cholesterol + 0.5% bile salts (to promote lipid absorption); at the same time, a control group was established using standard diet to test whether the model was successfully established. Monitoring frequency: Weigh twice a week (at a fixed time, after fasting for 4 hours); The criteria for successfully establishing an obesity model are: body weight exceeding that of the control group by more than 20% and serum TG > 1.5 mmol / L (the control group is about 1.0 mmol / L).

[0027] 2. Probiotic intervention experiment; Blank control group: standard feed + physiological saline (1 mL / kg / d); Obesity model group: high-fat diet + physiological saline (1mL / kg / d); Positive control group: high-fat diet + orlistat (20 mg / kg / d); Probiotic intervention group: high-fat diet + probiotic compound beverage prepared in Examples 1-3 / Comparative Examples 1-3 (1g / kg / d).

[0028] 3. Indicator testing; Weight gain rate (%) = (W 终止 -W 初始 ) / W 初始 ×100%; Fat coefficient (%) = (Wet weight of adipose tissue / Final body weight) × 100%; Serum TG (enzymatic GPO-PAP method).

[0029] The results are shown in Table 1:

[0030] As shown in the table above, the weight gain rate, fat coefficient, and serum TG level of the three embodiments were also significantly lower than those of the obesity model group, indicating that the probiotic compound drink has a significant weight loss effect, and the effect is better than that of orlistat. The weight gain rate, fat coefficient and serum TG level of Comparative Example 1 (without added plum fermentation powder) were higher than those of Examples 1-3, indicating that the addition of plum fermentation powder has a significant synergistic effect on weight loss, which may be related to the active ingredients (such as polyphenols) promoting fat metabolism. The weight gain rate, fat coefficient, and serum TG level of Comparative Example 2 (with unreasonable strain ratio) were higher than those of Examples 1-3, indicating that the strain ratio is crucial to the weight loss effect, and a reasonable ratio can enhance the colonization and metabolic function of probiotics. Comparative Example 3 (with an unreasonable strain ratio and no added plum fermentation powder) showed the worst results, verifying the synergistic effect of strain ratio and synergistic excipients on weight loss.

[0031] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a probiotic compound beverage with weight loss function, characterized in that, Includes the following steps: S1. Raw material pretreatment and modification; Soak the green plums in an aqueous solution of chlorine dioxide, rinse them with clean water, dry them, and then crush them. S2. Construct the fermentation system; Kluyveromycin was cultured at 28-32℃ for 12-18 hours. Lactobacillus helveticus was cultured at 30-35℃ for 18-24 hours; Acetobacter pasteurellium was cultured at 32-35℃ for 24-36 hours; The fermentation system was obtained by mixing Kluyveromyces martensii, Lactobacillus helveticus, and Acetobacter pasteurella multocida in a volume ratio of 3-5:2-4:1-3. S3. Fermentation; The crushed plum powder is mixed with syrup and introduced into the fermentation system. In the early stage of fermentation, the temperature is controlled at 25-28℃ for anaerobic fermentation for 2-3 days. The temperature is then raised to 30-33℃, and air containing carbon dioxide is introduced for micro-aerobic fermentation for 3-5 days. Ultrasonic fermentation is used to assist fermentation throughout the entire process. S4. Filtration and concentration; After fermentation, ultrafiltration membranes are used to remove microbial cells and macromolecular impurities from the fermentation broth to obtain a clarified fermentation broth. The fermentation broth is then vacuum concentrated to 1 / 5-1 / 8 of its original volume and freeze-dried to obtain plum fermentation powder. S5. Preparation of synergistic excipients; Add the green plum baking powder to the resistant dextrin and continue stirring to obtain the synergistic excipient; S6. Prepare a prebiotic solution; Weigh out 3-5 parts of inulin, 2-4 parts of fructooligosaccharides, and 1-2 parts of xylooligosaccharides, add them to 100 parts of warm water at 40-45℃, stir well and let them dissolve completely to obtain a prebiotic solution. S7. Preparation of microbial agents; Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were each inoculated into the culture medium and cultured to the logarithmic growth phase. The culture temperature for Lactobacillus gasseri is 30-37℃, and the pH is 6.0-7.

0. The culture temperature for Lactobacillus acidophilus is 35-38℃, and the pH is 5.5-6.

0. The culture temperature for Lactobacillus johnsonii and Lactobacillus plantarum is 30-35℃, and the pH is 6.5-7.

5. The culture temperature for Bifidobacterium adolescentis is 36-38℃, and the pH is 6.5-7.

2. The culture temperature for *Lactobacillus paracasei* is 37-40℃, and the pH is 6.8-7.

5. The cultured bacterial solution was centrifuged to separate the bacterial cells, which were then washed 2-3 times with sterile water. The bacterial cells were then freeze-dried to obtain freeze-dried bacterial powder of each strain. Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei were mixed evenly in a mass ratio of 1-3:2-4:2-4:3-5:2-5:2-3 to obtain the bacterial agent; The total number of viable bacteria in the bacterial agent is greater than or equal to 10. 9 CFU / g; S8. Mixing; Add 2-4 parts of bacterial agent, 8-11 parts of prebiotic solution, and 1-3 parts of synergistic excipients to a mixing tank, stir well and dry to obtain a probiotic compound drink with weight loss function.

2. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, S1 specifically involves: soaking the washed green plums in an aqueous solution containing 0.1-0.3% chlorine dioxide for 15-25 minutes, rinsing them thoroughly with clean water, and then drying them using vacuum freeze-drying technology. Under a vacuum of 0.01-0.03 MPa and a temperature of -30℃ to -40℃, the moisture content of the green plums is reduced to 10-15%. Finally, the dried green plums are pulverized into ultrafine powder with a particle size of 5-10 μm using an air jet mill.

3. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, In S2, the culture medium used to cultivate Kluyveromyces martensii consists of 30-60 g / L glucose, 10-30 g / L lactose, 12-25 g / L yeast extract, 5-15 g / L peptone, 2-5 g / L KH2PO4, 0.5-1.5 g / L MgSO4·7H2O, 1-3 g / L NaCl, 10-50 μg / L vitamin B7, 5-20 mg / L vitamin B3, 5-15 g / L mannitol, and 1-5 g / L wheat peptides.

4. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, In S2, the culture medium used to cultivate Lactobacillus helveticus consists of 20-40 g / L lactose, 10-20 g / L glucose, 15-25 g / L yeast extract, 5-10 g / L yeast peptone, 0.05-0.08 mol / L sodium citrate, 0.5-1.5 g / L MgSO4·7H2O, 0.05-0.15 g / L MgSO4·H2O, 5-20 mg / L vitamin B3, 5-15 g / L mannitol, and 0.5-2 g / L L-cysteine ​​hydrochloride.

5. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, In S2, the culture medium used to cultivate *Acetobacter pasteurellosis* consists of 30-45 g / L anhydrous ethanol, 5-15 g / L glucose, 20-35 g / L yeast extract, 5-12 g / L corn steep liquor powder, 3-8 g / L sodium citrate, 1-3 g / L betaine, 0.8-1.5 g / L MgSO4·7H2O, 2-4 g / L KH2PO4, 0.05-0.15 g / L MnSO4·H2O, 10-30 mg / L vitamin B5, 5-15 mg / L para-aminobenzoic acid, 0.5-2 g / L wheat oligopeptide, and 1-3 g / L trehalose.

6. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, S3 specifically involves mixing crushed plum powder with a high-concentration syrup of 30-40% by mass, with a syrup-to-plum powder mass ratio of 3-5:

1. This mixture is then introduced into the fermentation system, with the inoculation amount accounting for 5-10% of the total volume of the fermentation system. In the initial stage of fermentation, the temperature is controlled at 25-28℃ for anaerobic fermentation for 2-3 days. The temperature is then raised to 30-33℃, and filtered, sterilized air containing 5-10% carbon dioxide is introduced for micro-aerobic fermentation for 3-5 days. Throughout the fermentation process, ultrasonic waves are used to assist fermentation, with ultrasonic treatment performed every 4-6 hours. The ultrasonic frequency is 20-40kHz, the power is 100-200W, and the treatment time is 15-25 minutes.

7. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, S5 specifically involves: adding plum fermentation powder into a mixing tank to obtain a mixed system; first, setting the stirring speed to 60-80 r / min; heating to 55-60℃ at a rate of 1-3℃ / min; continuing to stir at this temperature for 30-35 min; then cooling to 40-45℃ at a rate of 2-4℃ / min; adding 4-6.5% resistant dextrin by mass of the plum fermentation powder; and continuing to stir for 25-30 min to obtain the synergistic excipient.

8. The preparation method of the probiotic compound beverage with weight loss function according to claim 1, characterized in that, In S7, the culture medium used to cultivate Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus johnsonii, Lactobacillus plantarum, Bifidobacterium adolescentis, and Lactobacillus paracasei consists of 8-12 parts peptone, 6-10 parts beef extract, 3-5 parts yeast extract, 18-22 parts glucose, 0.1-0.3 parts magnesium sulfate, 4-6 parts sodium acetate, 1-3 parts diammonium citrate, 1-3 parts dipotassium hydrogen phosphate, 0.03-0.05 parts manganese sulfate, 0.8-1.2 parts Tween 80, and 1000 parts water.

9. A probiotic compound beverage with weight loss function prepared by the preparation method of any one of claims 1-8.

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