Saccharomyces cerevisiae probiotic preparation formula and preparation process thereof

Abstract

The invention relates to the technical field of microbial preparations, and discloses a saccharomyces cerevisiae probiotic preparation formula and a preparation process thereof, and the saccharomyces cerevisiae probiotic preparation formula comprises the following raw materials in parts by mass: 15-30 parts of fructo-oligosaccharide, 10-20 parts of blueberry fruit powder, 5-15 parts of saccharomyces cerevisiae CNCMI-3799, 10-25 parts of maltodextrin, 3-8 parts of xylooligosaccharide, 5-10 parts of erythritol, and 10-20 parts of probiotic powder. The product is stable in quality and uniform in particle shape, has good fluidity, is adaptive to automatic pressing and packaging production, improves the production efficiency, meets the requirements of large-scale and standardized production, and integrally realizes collaborative improvement of product functions, quality and production suitability.

Description

Technical Field

[0001] This invention relates to the field of microbial preparation technology, specifically to the formulation and preparation process of a Saccharomyces cerevisiae probiotic preparation. Background Technology

[0002] Probiotic preparations are widely used in the health field, offering numerous health benefits such as regulating gut microbiota balance, enhancing immunity, and promoting nutrient absorption. However, current probiotic preparations on the market face several pressing issues. During the raw material mixing process, differences in particle size, density, and other physical properties among the raw materials lead to uneven mixing and inconsistent distribution of components within the product, affecting the stability and uniformity of its efficacy. Existing products often use single probiotic strains, which have limited intestinal colonization capabilities, thus restricting product functionality. Traditional preparation methods often employ mixing processes, resulting in uneven raw material particle size, stratification, and clumping during storage. This not only affects the efficiency of automated packaging but also leads to the inactivation of live bacteria, further reducing product quality.

[0003] In addition, the existing process is not well adapted to the characteristics of raw materials, resulting in large losses of microbial communities during the production process and large fluctuations in the quality of the final product, which makes it difficult to meet the needs of large-scale and standardized production. There is an urgent need for a method that takes into account both high activity of microorganisms and stable granulation. Summary of the Invention

[0004] (a) Technical problems to be solved

[0005] To address the shortcomings of existing technologies, this invention provides a formula for a brewing yeast probiotic preparation and its preparation process.

[0006] (II) Technical Solution

[0007] To achieve the above objectives, the present invention provides the following technical solution: a brewer's yeast probiotic preparation formulation, comprising the following raw materials in parts by weight:

[0008] 15-30 parts of fructooligosaccharides, 10-20 parts of blueberry fruit powder, 5-15 parts of brewer's yeast CNCMI-3799, 10-25 parts of maltodextrin, 3-8 parts of xylooligosaccharides, 5-10 parts of erythritol, and 10-20 parts of probiotic powder.

[0009] The activated Saccharomyces cerevisiae CNCMI-3799 is used in the production of Saccharomyces cerevisiae probiotic preparations.

[0010] The activation process of the brewing yeast CNCMI-3799 includes the following steps:

[0011] A1. Accurately weigh the culture medium raw materials, pour them into a sterile activation tank, add purified water and stir, then add purified water to make up to the total amount to obtain the culture medium. Sterilize by autoclaving and set aside.

[0012] A2. Take the freeze-dried Saccharomyces cerevisiae CNCMI-3799 and inoculate it into the culture medium. Turn on the stirrer, start the temperature control system, turn on the ventilation device, introduce sterile air, and culture to obtain the seed culture.

[0013] A3. Add the culture medium from step A1 to the sterile activation tank, transfer the seed liquid from step A2 into the activation tank, add maltodextrin during the culture, and continue the culture to obtain yeast liquid.

[0014] A4. Transfer the yeast liquid from step A3 into a sterile centrifugal concentrator to concentrate it, and collect the yeast concentrate at the bottom.

[0015] A5. The yeast concentrate is vacuum freeze-dried, pulverized at low temperature, sieved through a 40-mesh sterile sieve, and the sieve-underfill material is collected, which is the activated brewing yeast CNCMI-3799.

[0016] Furthermore, the probiotic powder contains 10-30% Lactobacillus rhamnosus, 15-30% Bifidobacterium animalis, 10-30% Lactobacillus paracasei, 10-30% Lactobacillus plantarum, and 10-20% Lactobacillus acidophilus.

[0017] Furthermore, the brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder are all 40-mesh granules, with live cell counts of brewing yeast CNCMI-3799 ≥ 1 × 10⁻⁶. 10 CFU / g, probiotic powder ≥5×10 10 CFU / g.

[0018] Furthermore, the culture medium ingredients consist of 180–200 parts glucose, 50–80 parts yeast extract, 10–15 parts galactooligosaccharides, 8–12 parts maltodextrin, 1–1.5 parts potassium citrate chelated zinc, 5–8 parts wheat protein peptides, and 0.001–0.002 parts vitamin B. 12 .

[0019] Furthermore, the preparation method of potassium citrate chelated zinc includes the following steps:

[0020] B1. Weigh 180-220 parts of citric acid, add 800-1000 parts of purified water, place on a constant temperature magnetic stirrer, stir at 30-40℃ until completely dissolved, and rotate at 300-500 r / min to obtain a citric acid solution; weigh 80-85 parts of zinc carbonate, slowly add to the above citric acid solution, and continue stirring at 30-35℃ for 20-30 min until no bubbles are generated to obtain a transparent zinc citrate solution;

[0021] B2. Add 160-180 parts of potassium citrate to the zinc citrate solution, heat to 45-50℃, adjust the stirring speed to 500-800 r / min, adjust the pH to 5.5-6.0, and continue the reaction for 2-3 hours.

[0022] B3. Transfer the above reaction solution into a vacuum rotary evaporator, set the temperature to 60-80℃ and the vacuum degree to 0.06-0.08MPa, and concentrate it to 1 / 5 of the original volume to obtain a concentrated solution;

[0023] B4. Pour the concentrated liquid into a freeze-drying tray, place it in a freeze dryer, pre-freeze at -20 to -40°C for 2 to 5 hours, maintain a vacuum of 5 to 10 Pa and a sublimation temperature of 30 to 40°C, dry for 8 to 12 hours, pulverize with a pulverizer at a speed of 1000 r / min for 3 minutes, and pass through a 40-mesh sieve to obtain potassium citrate chelated zinc.

[0024] Further, the activation tank parameters for step A2 are: rotation speed 80-100 r / min, temperature 30-32℃, and air flow rate 0.5-1 L / (L·min); the activation tank parameters for step A3 are: rotation speed 80-100 r / min, temperature 30-35℃, and air flow rate 0.8-1.2 L / (L·min).

[0025] Furthermore, the preparation method of the Saccharomyces cerevisiae probiotic preparation includes the following steps:

[0026] S1. Grind and sieve the brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder to a 40-mesh standard.

[0027] S2. Add fructooligosaccharides, maltodextrin, xylooligosaccharides, erythritol, blueberry fruit powder, activated brewer's yeast CNCMI-3799, and probiotic powder to a mixer and stir to obtain a mixture. Set the mixing time to 20-30 min and the mixing speed to 20-30 r / min.

[0028] S3. Select a vertical fluidized bed granulator, put the mixture into the fluidized bed of the fluidized bed granulator, turn on the air intake system, and after the temperature inside the bed stabilizes to the set range, start the atomizing nozzle to spray purified water. When the particle size initially reaches 30-40 mesh, stop spraying and continue ventilation for 3 minutes.

[0029] S4. Place the granulated material into a hot air drying oven, set the drying temperature to 50-60℃, and the drying time to 1-2 hours. Pass the material through a 40-mesh sieve and feed the sieved granules into a horizontal mixer. Add an anti-caking agent at 0.4-0.6% of the total granule weight, set the rotation speed to 80-100 r / min, and mix for 5-8 minutes. After mixing, package the material using a fully automatic strip bag packaging machine. The parameters are as follows: bag type is a three-side seal strip bag, size is 10cm×6cm, heat sealing temperature is 120-130℃, heat sealing time is 0.8s, filling speed is 40-50 bags / min, and net content is 3g per bag.

[0030] Furthermore, in step S3, the process parameters for the vertical fluidized bed granulator are set as follows: inlet air temperature 60-65℃, outlet air temperature 35-40℃, atomization pressure 0.22-0.25MPa, spraying speed 6-8mL / min, and fluidized bed air velocity 1.2-1.5m / s.

[0031] Further, in step S4, the anti-caking agent is one of fumed silica, magnesium stearate, and tricalcium phosphate; the fumed silica has a particle size of 5-10 μm and a purity of ≥99.8%; the magnesium stearate has a particle size of 10-20 μm and a purity of ≥99%; the tricalcium phosphate has a particle size of 20-30 μm, a calcium content of ≥38%, and a phosphorus content of ≥18%.

[0032] (III) Beneficial Technical Effects

[0033] This invention pretreats brewer's yeast through an activation process, activating the strain's metabolic activity and enhancing its resistance. This allows the strain to better tolerate temperature and humidity changes during subsequent processing and storage, reducing activity loss. The stable cell morphology formed during activation also lays the foundation for uniform mixing in subsequent formulations. A combination of multiple probiotics is used, leveraging the synergistic effects between different strains to enhance intestinal colonization. Prebiotics not only provide nutritional support for probiotics but also regulate the intestinal microenvironment, promoting the proliferation of beneficial bacteria. Combined with natural flavor substances, the product's taste is improved while maintaining functionality, enhancing consumer acceptance. The invention incorporates fluidized bed granulation technology, precisely controlling granulation parameters to form uniform particles, avoiding the stratification problems caused by traditional simple mixing and improving overall product uniformity. Simultaneously, a suitable anti-caking agent is added to form a physical barrier or hydrophobic layer on the particle surface, reducing particle adhesion and effectively solving the clumping problem during storage. Furthermore, the appropriate selection of anti-caking agents does not affect the viable bacterial activity.

[0034] In addition, the entire process fully considers the characteristics of raw materials, optimizes temperature parameters for heat-sensitive microorganisms, and adjusts the processing methods for hygroscopic raw materials to ensure minimal additional loss of live bacteria during production and stable product quality. At the same time, the uniform particle shape and good flowability are suitable for automated compression packaging production, improving production efficiency and meeting the needs of large-scale and standardized production, thus achieving a synergistic improvement in product function, quality and production adaptability. Detailed Implementation

[0035] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0036] Unless otherwise specified, all components of the brewer's yeast probiotic preparation of this invention are commercially available.

[0037] The parts used in this invention are parts by weight, and the percentages are percentages by weight.

[0038] Example 1

[0039] The formula for a brewer's yeast probiotic preparation includes the following ingredients by weight:

[0040] 15 parts fructooligosaccharides, 10 parts blueberry fruit powder, 5 parts brewer's yeast CNCMI-3799, 10 parts maltodextrin, 3 parts xylooligosaccharides, 5 parts erythritol, and 10 parts probiotic powder.

[0041] The activated Saccharomyces cerevisiae CNCMI-3799 is used in the production of Saccharomyces cerevisiae probiotic preparations.

[0042] The probiotic powder contains 30% Lactobacillus rhamnosus, 30% Bifidobacterium animalis, 10% Lactobacillus paracasei, 20% Lactobacillus plantarum, and 10% Lactobacillus acidophilus.

[0043] The brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder are all 40-mesh granules, with live bacteria counts of brewing yeast CNCMI-3799 ≥ 1 × 10⁻⁶. 10 CFU / g, probiotic powder ≥5×10 10 CFU / g.

[0044] The culture medium consisted of 180 parts glucose, 50 parts yeast extract, 10 parts galacto-oligosaccharides, 8 parts maltodextrin, 1 part potassium citrate chelated zinc, 5 parts wheat protein peptides, and 0.001 parts vitamin B. 12 .

[0045] The activation process of CNCMI-3799 brewing yeast includes the following steps:

[0046] A1. Outside the aseptic operating table, accurately weigh the culture medium raw materials and pour them into the aseptic activation tank. Add 8000 parts of purified water, turn on the stirring function of the activation tank, and stir at 50 r / min for 10 min. Add purified water to make up the solution in the tank to 10000 parts, and continue stirring for 5 min to obtain the culture medium. Perform high-pressure steam sterilization, setting the parameters to 121℃ and 0.1MPa for 20 min. After sterilization, introduce sterile air to cool down. When the temperature inside the tank drops to 30℃, close the vent valve and set it aside for later use.

[0047] A2. Sterilize the aseptic workbench with UV for 20 minutes, then turn off the UV and ventilate for 10 minutes. Use a sterile inoculation gun to draw 10 portions of freeze-dried Saccharomyces cerevisiae CNCMI-3799 and quickly insert it into the inlet of the activation tank to inoculate all the powder into the culture medium. Immediately close the inlet, turn on the stirrer, set the speed to 80 r / min, start the temperature control system to maintain a constant temperature of 30℃ inside the tank, turn on the ventilation device to introduce sterile air, control the ventilation rate to 0.5 L / (L·min), and continue culturing for 4 hours to obtain the seed culture.

[0048] A3. Add the culture medium from step A1 to the aseptic activation tank, and transfer the seed liquid from step A2 into the activation tank through a sterile pipeline. Set the rotation speed to 80 r / min, maintain the temperature at 30℃, the aeration rate to 0.8 L / (L·min), and the incubation time to 2 h. When the incubation reaches 1.5 h, add 50 parts of maltodextrin to the activation tank under aseptic conditions and continue the incubation to obtain yeast liquid.

[0049] A4. Transfer the yeast liquid from step A3 into a sterile centrifugal concentrator, set the temperature to 4℃ and the speed to 3000r / min, start centrifugation and continue for 10 minutes. After centrifugation, collect the yeast concentrate at the bottom.

[0050] A5. Spread the yeast concentrate evenly on a sterile freeze-drying tray, place it in a vacuum freeze dryer, freeze at -20℃ for 5 hours, then maintain a vacuum of 10Pa and a sublimation drying temperature of 30℃ for 8 hours. Put the dried yeast powder into a low-temperature pulverizer, set the pulverizing temperature to ≤30℃ and the rotation speed to 1200r / min, pulverize for 5 minutes, and sieve through a 40-mesh sterile sieve. Collect the sieve material, which is the activated brewing yeast CNCMI-3799.

[0051] The preparation method of potassium citrate chelated zinc includes the following steps:

[0052] B1. Weigh 180 parts of citric acid, add 800 parts of purified water, place on a constant temperature magnetic stirrer, stir at 30°C until completely dissolved, and rotate at 300 r / min to obtain a citric acid solution; weigh 80 parts of zinc carbonate, slowly add to the above citric acid solution, continue stirring at 30°C for 20 min until no bubbles are generated, to obtain a transparent zinc citrate solution.

[0053] B2. Add 160 parts of potassium citrate to the zinc citrate solution, heat to 45°C, adjust the stirring speed to 500 r / min, adjust the pH to 5.5, and continue the reaction for 2 hours.

[0054] B3. Transfer the above reaction solution into a vacuum rotary evaporator, set the temperature to 60℃ and the vacuum degree to 0.06MPa, and concentrate it to 1 / 5 of the original volume to obtain a concentrated solution.

[0055] B4. Pour the concentrate into a freeze-drying tray, place it in a freeze dryer, pre-freeze at -20℃ for 2 hours, maintain a vacuum of 5Pa and a sublimation temperature of 30℃, dry for 8 hours, pulverize with a pulverizer at a speed of 1000r / min for 3 minutes, and pass through a 40-mesh sieve to obtain potassium citrate chelated zinc.

[0056] The preparation method of the Saccharomyces cerevisiae probiotic preparation includes the following steps:

[0057] S1. Grind and sieve the brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder to a 40-mesh standard.

[0058] S2. Add fructooligosaccharides, maltodextrin, xylooligosaccharides, erythritol, blueberry fruit powder, activated brewer's yeast CNCMI-3799 and probiotic powder to a mixer and stir to obtain a mixture. Set the mixing time to 20 min and the mixing speed to 20 r / min.

[0059] S3. Select a vertical fluidized bed granulator, put the mixture into the fluidized bed of the fluidized bed granulator, turn on the air intake system, and after the temperature inside the bed stabilizes to the set range, start the atomizing nozzle to spray purified water. When the particle size initially reaches 30-40 mesh, stop spraying and continue ventilation for 3 minutes.

[0060] S4. Place the granulated material into a hot air drying oven, set the drying temperature to 50℃, and the drying time to 1 hour. Pass the material through a 40-mesh sieve and then feed the sieved granules into a horizontal mixer. Add an anti-caking agent at 0.4% of the total granule weight, set the rotation speed to 80 r / min, and mix for 5 minutes. After mixing, package the material using a fully automatic strip bag packaging machine. The parameters are as follows: bag type is a three-side seal strip bag, size is 10cm × 6cm, heat sealing temperature is 120℃, heat sealing time is 0.8s, filling speed is 40 bags / min, and net content per bag is 3g.

[0061] In step S3, the process parameters for the vertical fluidized bed granulator are set as follows: inlet air temperature 60℃, outlet air temperature 35℃, atomization pressure 0.22MPa, spraying speed 6mL / min, and fluidized bed air velocity 1.2m / s.

[0062] In step S4, the anti-caking agent is fumed silica; the fumed silica has a particle size of 5-10 μm and a purity of ≥99.8%.

[0063] Example 2

[0064] The formula for a brewer's yeast probiotic preparation includes the following ingredients by weight:

[0065] 20 parts fructooligosaccharides, 15 parts blueberry fruit powder, 10 parts brewer's yeast CNCMI-3799, 20 parts maltodextrin, 5 parts xylooligosaccharides, 8 parts erythritol, and 15 parts probiotic powder.

[0066] The activated Saccharomyces cerevisiae CNCMI-3799 is used in the production of Saccharomyces cerevisiae probiotic preparations.

[0067] The probiotic powder contains 20% Lactobacillus rhamnosus, 20% Bifidobacterium animalis, 30% Lactobacillus paracasei, 10% Lactobacillus plantarum, and 20% Lactobacillus acidophilus.

[0068] The brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder are all 40-mesh granules, with live bacteria counts of brewing yeast CNCMI-3799 ≥ 1 × 10⁻⁶. 10 CFU / g, probiotic powder ≥5×10 10 CFU / g.

[0069] The culture medium consisted of 190 parts glucose, 60 parts yeast extract, 12 parts galacto-oligosaccharides, 10 parts maltodextrin, 1.5 parts potassium citrate chelated zinc, 6 parts wheat protein peptides, and 0.001 parts vitamin B. 12 .

[0070] The activation process of CNCMI-3799 brewing yeast includes the following steps:

[0071] A1. Outside the aseptic operating table, accurately weigh the culture medium raw materials and pour them into the aseptic activation tank. Add 8000 parts of purified water, turn on the stirring function of the activation tank, and stir at 80 r / min for 10 min. Add purified water to make up the solution in the tank to 10000 parts, and continue stirring for 5 min to obtain the culture medium. Perform high-pressure steam sterilization, setting the parameters to 121℃ and 0.1MPa for 20 min. After sterilization, introduce sterile air to cool down. When the temperature inside the tank drops to 30℃, close the vent valve and set it aside for later use.

[0072] A2. Sterilize the aseptic workbench with UV for 25 minutes, then turn off the UV and ventilate for 10 minutes. Use a sterile inoculation gun to draw 10 portions of freeze-dried Saccharomyces cerevisiae CNCMI-3799 and quickly insert it into the inlet of the activation tank to inoculate all the powder into the culture medium. Immediately close the inlet, turn on the stirrer, set the speed to 90 r / min, start the temperature control system to maintain a constant temperature of 30℃ inside the tank, turn on the ventilation device to introduce sterile air, control the ventilation rate to 0.8 L / (L·min), and continue culturing for 4.5 h to obtain the seed culture.

[0073] A3. Add the culture medium from step A1 to the aseptic activation tank, and transfer the seed liquid from step A2 into the activation tank through a sterile pipeline. Set the rotation speed to 90 r / min, maintain the temperature at 32℃, the aeration rate to 1 L / (L·min), and the incubation time to 2.5 h. After 1.5 h of incubation, add 50 parts of maltodextrin to the activation tank under aseptic conditions and continue incubation to obtain yeast liquid.

[0074] A4. Transfer the yeast liquid from step A3 into a sterile centrifugal concentrator, set the temperature to 5℃ and the speed to 4000r / min, start the centrifugation and continue for 10 minutes. After centrifugation, collect the yeast concentrate at the bottom.

[0075] A5. Spread the yeast concentrate evenly on a sterile freeze-drying tray, place it in a vacuum freeze dryer, freeze at -30℃ for 4 hours, then maintain a vacuum of 15Pa and a sublimation drying temperature of 32℃ for 9 hours. Put the dried yeast powder into a low-temperature pulverizer, set the pulverizing temperature to ≤30℃ and the rotation speed to 1300r / min, pulverize for 6 minutes, and sieve through a 40-mesh sterile sieve. Collect the sieve material, which is the activated brewing yeast CNCMI-3799.

[0076] The preparation method of potassium citrate chelated zinc includes the following steps:

[0077] B1. Weigh 200 parts of citric acid, add 900 parts of purified water, place on a constant temperature magnetic stirrer, stir at 35°C until completely dissolved, and rotate at 400 r / min to obtain a citric acid solution; weigh 82 parts of zinc carbonate, slowly add to the above citric acid solution, continue stirring at 32°C for 25 min until no bubbles are generated, to obtain a transparent zinc citrate solution.

[0078] B2. Add 170 parts of potassium citrate to the zinc citrate solution, heat to 45°C, adjust the stirring speed to 600 r / min, adjust the pH to 5.8, and continue the reaction for 2.5 h.

[0079] B3. Transfer the above reaction solution into a vacuum rotary evaporator, set the temperature to 70℃ and the vacuum degree to 0.07MPa, and concentrate it to 1 / 5 of the original volume to obtain a concentrated solution.

[0080] B4. Pour the concentrate into a freeze-drying tray, place it in a freeze dryer, pre-freeze at -30℃ for 3 hours, maintain a vacuum of 6Pa and a sublimation temperature of 35℃, dry for 10 hours, pulverize with a pulverizer at a speed of 1000r / min for 3 minutes, and pass through a 40-mesh sieve to obtain potassium citrate chelated zinc.

[0081] The preparation method of the Saccharomyces cerevisiae probiotic preparation includes the following steps:

[0082] S1. Grind and sieve the brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder to a 40-mesh standard.

[0083] S2. Add fructooligosaccharides, maltodextrin, xylooligosaccharides, erythritol, blueberry fruit powder, activated brewer's yeast CNCMI-3799 and probiotic powder to a mixer and stir to obtain a mixture. Set the mixing time to 25 min and the mixing speed to 25 r / min.

[0084] S3. Select a vertical fluidized bed granulator, put the mixture into the fluidized bed of the fluidized bed granulator, turn on the air intake system, and after the temperature inside the bed stabilizes to the set range, start the atomizing nozzle to spray purified water. When the particle size initially reaches 30-40 mesh, stop spraying and continue ventilation for 3 minutes.

[0085] S4. Place the granulated material into a hot air drying oven, set the drying temperature to 55℃, and the drying time to 1 hour. Pass the material through a 40-mesh sieve and then feed the sieved granules into a horizontal mixer. Add an anti-caking agent at 0.5% of the total granule weight, set the rotation speed to 90 r / min, and mix for 6 minutes. After mixing, package the material using a fully automatic strip bag packaging machine. The parameters are as follows: bag type is a three-side seal strip bag, size is 10cm × 6cm, heat sealing temperature is 125℃, heat sealing time is 0.8s, filling speed is 45 bags / min, and net content per bag is 3g.

[0086] In step S3, the process parameters for the vertical fluidized bed granulator are set as follows: inlet air temperature 62℃, outlet air temperature 38℃, atomization pressure 0.24MPa, spraying speed 7mL / min, and fluidized bed air velocity 1.4m / s.

[0087] In step S4, the anti-caking agent is magnesium stearate; the magnesium stearate has a particle size of 10-20 μm and a purity of ≥99%.

[0088] Example 3

[0089] The formula for a brewer's yeast probiotic preparation includes the following ingredients by weight:

[0090] 30 parts fructooligosaccharides, 20 parts blueberry fruit powder, 15 parts brewer's yeast CNCMI-3799, 25 parts maltodextrin, 8 parts xylooligosaccharides, 10 parts erythritol, and 20 parts probiotic powder.

[0091] The activated Saccharomyces cerevisiae CNCMI-3799 is used in the production of Saccharomyces cerevisiae probiotic preparations.

[0092] The probiotic powder contains 20% Lactobacillus rhamnosus, 30% Bifidobacterium animalis, 30% Lactobacillus paracasei, 10% Lactobacillus plantarum, and 10% Lactobacillus acidophilus.

[0093] The brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder are all 40-mesh granules, with live bacteria counts of brewing yeast CNCMI-3799 ≥ 1 × 10⁻⁶. 10 CFU / g, probiotic powder ≥5×10 10 CFU / g.

[0094] The culture medium consisted of 200 parts glucose, 80 parts yeast extract, 15 parts galacto-oligosaccharides, 12 parts maltodextrin, 1.5 parts potassium citrate chelated zinc, 8 parts wheat protein peptides, and 0.002 parts vitamin B. 12 .

[0095] The activation process of CNCMI-3799 brewing yeast includes the following steps:

[0096] A1. Outside the aseptic operating table, accurately weigh the culture medium raw materials and pour them into the aseptic activation tank. Add 8000 parts of purified water, turn on the stirring function of the activation tank, and stir at 100 r / min for 15 min. Add purified water to make up the solution in the tank to 10000 parts, and continue stirring for 10 min to obtain the culture medium. Perform high-pressure steam sterilization, setting the parameters to 121℃ and 0.1MPa for 20 min. After sterilization, introduce sterile air to cool down. When the temperature inside the tank drops to 32℃, close the vent valve and set it aside for later use.

[0097] A2. Sterilize the aseptic operating table with UV for 30 minutes, then turn off the UV and ventilate for 10 minutes. Use a sterile inoculation gun to take 10 portions of freeze-dried Saccharomyces cerevisiae CNCMI-3799 and quickly insert it into the inlet of the activation tank to inoculate all the powder into the culture medium. Immediately close the inlet, turn on the stirrer, set the speed to 100 r / min, start the temperature control system to maintain a constant temperature of 32℃ inside the tank, turn on the ventilation device to introduce sterile air, control the ventilation rate to 1L / (L·min), and continue to culture for 5 hours to obtain the seed culture.

[0098] A3. Add the culture medium from step A1 to the aseptic activation tank, and transfer the seed liquid from step A2 into the activation tank through a sterile pipeline. Set the rotation speed to 100 r / min, maintain the temperature at 35℃, the aeration rate to 1.2 L / (L·min), and the incubation time to 3 h. When the culture reaches 1.5 h, add 50 parts of maltodextrin to the activation tank under aseptic conditions and continue the culture to obtain the yeast liquid.

[0099] A4. Transfer the yeast liquid from step A3 into a sterile centrifugal concentrator, set the temperature to 6℃ and the speed to 5000r / min, start the centrifugation and continue for 15min. After centrifugation, collect the yeast concentrate at the bottom.

[0100] A5. Spread the yeast concentrate evenly on a sterile freeze-drying tray, place it in a vacuum freeze dryer, freeze at -40℃ for 5 hours, then maintain a vacuum of 20Pa and a sublimation drying temperature of 35℃ for 10 hours. Put the dried yeast powder into a low-temperature pulverizer, set the pulverizing temperature to ≤30℃ and the rotation speed to 1500r / min, pulverize for 8 minutes, and sieve through a 40-mesh sterile sieve. Collect the sieve material, which is the activated brewing yeast CNCMI-3799.

[0101] The preparation method of potassium citrate chelated zinc includes the following steps:

[0102] B1. Weigh 220 parts of citric acid, add 1000 parts of purified water, place on a constant temperature magnetic stirrer, stir at 40°C until completely dissolved, and rotate at 500 r / min to obtain a citric acid solution; weigh 85 parts of zinc carbonate, slowly add to the above citric acid solution, continue stirring at 35°C for 30 min until no bubbles are generated, to obtain a transparent zinc citrate solution.

[0103] B2. Add 180 parts of potassium citrate to the zinc citrate solution, heat to 50°C, adjust the stirring speed to 800 r / min, adjust the pH to 6.0, and continue the reaction for 3 hours.

[0104] B3. Transfer the above reaction solution into a vacuum rotary evaporator, set the temperature to 80℃ and the vacuum degree to 0.08MPa, and concentrate it to 1 / 5 of the original volume to obtain a concentrated solution.

[0105] B4. Pour the concentrate into a freeze-drying tray, place it in a freeze dryer, pre-freeze at -40℃ for 5 hours, maintain a vacuum of 10Pa and a sublimation temperature of 40℃, dry for 12 hours, pulverize with a pulverizer at a speed of 1000r / min for 3 minutes, and pass through a 40-mesh sieve to obtain potassium citrate chelated zinc.

[0106] The preparation method of the Saccharomyces cerevisiae probiotic preparation includes the following steps:

[0107] S1. Grind and sieve the brewing yeast CNCMI-3799, probiotic powder, and blueberry fruit powder to a 40-mesh standard.

[0108] S2. Add fructooligosaccharides, maltodextrin, xylooligosaccharides, erythritol, blueberry fruit powder, activated brewer's yeast CNCMI-3799 and probiotic powder to a mixer and stir to obtain a mixture. Set the mixing time to 30 min and the mixing speed to 30 r / min.

[0109] S3. Select a vertical fluidized bed granulator, put the mixture into the fluidized bed of the fluidized bed granulator, turn on the air intake system, and after the temperature inside the bed stabilizes to the set range, start the atomizing nozzle to spray purified water. When the particle size initially reaches 30-40 mesh, stop spraying and continue ventilation for 3 minutes.

[0110] S4. Place the granulated material into a hot air drying oven, set the drying temperature to 60℃, and the drying time to 2 hours. Pass the material through a 40-mesh sieve and then feed the sieved granules into a horizontal mixer. Add an anti-caking agent at 0.6% of the total granule weight, set the rotation speed to 100 r / min, and mix for 8 minutes. After mixing, package the material using a fully automatic strip bag packaging machine. The parameters are as follows: bag type is a three-side seal strip bag, size is 10cm × 6cm, heat sealing temperature is 130℃, heat sealing time is 0.8s, filling speed is 50 bags / min, and net content per bag is 3g.

[0111] In step S3, the process parameters for the vertical fluidized bed granulator are set as follows: inlet air temperature 65℃, outlet air temperature 40℃, atomization pressure 0.25MPa, spraying speed 8mL / min, and fluidized bed air velocity 1.5m / s.

[0112] In step S4, the anti-caking agent is tricalcium phosphate; the tricalcium phosphate has a particle size of 20-30 μm, a calcium content of ≥38%, and a phosphorus content of ≥18%.

[0113] Comparative Example 1: The brewing yeast CNCMI-3799 was used directly without activation treatment, and the remaining processes and parameters were the same as in Example 1.

[0114] Comparative Example 2: The probiotic powder used was a single Lactobacillus rhamnosus, and the other processes and parameters were the same as in Example 1.

[0115] Comparative Example 3: The fluidized bed granulation process in step S3 was removed, and the mixture from step S2 was directly dried, sieved, and packaged. No anti-caking agent was added in step S4. The remaining processes and parameters were the same as in Example 1.

[0116] Performance testing:

[0117] 1. Live bacteria survival rate: including the survival rate of live brewer's yeast and the total live probiotic survival rate (before storage + after 6 months of storage);

[0118] Reference standards: GB 4789.34-2016 (Yeast), GB 4789.35-2016 (Probiotics);

[0119] Test method: Plate count method: Take 10g of sample and serially dilute with sterile physiological saline (10... -1 -10 -8The bacteria were inoculated into YPD medium (yeast) and MRS medium (probiotics) respectively, and counted after being incubated at 37°C for 48 hours. The survival rate was calculated as (number of live bacteria after storage / number of live bacteria before storage) × 100%.

[0120] Table 1 Results of viable bacteria survival rate test

[0121]

[0122] 2. Particle uniformity

[0123] 40-mesh pass rate: GB / T 19284-2018; Take 100g of sample, pass it through a 40-mesh standard sieve (aperture 0.425mm), collect the sieve residue, and calculate the pass rate = (weight of sieve residue / total weight of sample) × 100%.

[0124] Mixing uniformity: GB / T 29889-2013; To detect the content of fructooligosaccharides (marker) in the sample, 5 sampling points are randomly selected, and the coefficient of variation CV = (standard deviation / mean) × 100% is calculated. The smaller the CV, the better the uniformity.

[0125] Table 2 Results of particle uniformity test

[0126] Group 40-mesh pass rate (%) Mixing uniformity (CV, %) Example 1 97.5 1.8 Example 2 98.2 1.5 Example 3 97.8 1.6 Comparative Example 1 97.9 1.7 Comparative Example 2 98.0 1.6 Comparative Example 3 75.3 5.2

[0127] 3. Anti-caking properties and flowability

[0128] Agglomeration rate: QB / T 4261-2011; Take 100g of sample stored for 6 months, pass it through a 40-mesh sieve, collect the agglomerates on the sieve, crush them and weigh them, and calculate the agglomeration rate = (weight of agglomerates / total weight of sample) × 100%.

[0129] Angle of repose: GB / T 16913-2008; The sample is allowed to fall freely through a funnel onto a horizontal table to form a cone. The height H and the bottom radius R of the cone are measured, and the angle of repose θ = arctan(2H / R) is calculated. θ ≤ 30° indicates excellent flowability.

[0130] Table 3 Results of Anti-caking and Flowability Tests

[0131] Group Clumping rate after 6 months of storage (%) Angle of repose (°) Example 1 1.2 27.5 Example 2 1.0 26.8 Example 3 1.3 28.2 Comparative Example 1 1.1 27.2 Comparative Example 2 1.2 27.0 Comparative Example 3 18.5 45.3

[0132] As shown in Table 1, Comparative Example 1, which did not undergo the activation process of brewing yeast, had a survival rate of 62.3% after 6 months, which was low. This indicates that the activation process can significantly improve the yeast's resistance to stress and reduce the loss of activity during storage. Comparative Example 2, which used a single Lactobacillus rhamnosus, had a survival rate of 75.2% after 6 months of probiotic storage, indicating that the combination of 5 probiotics can improve the overall stability of live bacteria through synergistic effects.

[0133] As shown in Tables 2 and 3, Comparative Example 3, without fluidized bed granulation and without anti-caking agent, achieved a 40-mesh pass rate of 75.3%, a mixing uniformity of 5.2%, a clumping rate of 18.5% after 6 months of storage, and an angle of repose of 45.3°. This indicates that fluidized bed granulation can ensure particle uniformity, and anti-caking agent can effectively solve the clumping problem while improving flowability, thus meeting the automated production requirements of compression packaging.

[0134] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A formulation of Saccharomyces cerevisiae probiotic bacteria characterized in that, The following ingredients are included by mass: 15-30 parts of fructooligosaccharide, 10-20 parts of blueberry fruit powder, 10-15 parts of Saccharomyces cerevisiae CNCMI-3799, 10-25 parts of malt dextrin, 3-8 parts of xylooligosaccharide, 5-10 parts of erythritol, and 10-20 parts of probiotic bacteria powder; The activated Saccharomyces cerevisiae CNCMI-3799 is used for the production of Saccharomyces cerevisiae probiotic bacteria preparation; The activation process of the Saccharomyces cerevisiae CNCMI-3799 includes the following steps: A1. Accurately weigh the culture medium raw materials, pour them into a sterile activation tank, add purified water and stir, then add purified water to make up the total amount to prepare the culture medium, high-pressure steam sterilize, and reserve for use; A2. Absorb the Saccharomyces cerevisiae CNCMI-3799 freeze-dried bacteria powder into the culture medium, start stirring, start the temperature control system, open the aeration device, and introduce sterile air to culture the seed liquid; A3. Add the culture medium of step A1 to the sterile activation tank, transfer the seed liquid of step A2 into the activation tank, add malt dextrin during the culture, and continue the culture to obtain the yeast liquid; A4. Transfer the yeast liquid of step A3 into a sterile centrifugal concentrator for concentration, and collect the yeast concentrate at the bottom; A5. Vacuum freeze-dry the yeast concentrate, low-temperature pulverize, sieve through a 40-mesh sterile screen, and collect the undersize material, which is the activated Saccharomyces cerevisiae CNCMI-3799.

2. The Saccharomyces cerevisiae probiotic formulation according to claim 1, characterized in that, The probiotic bacteria powder contains 10-30% of Lactobacillus rhamnosus, 15-30% of Bifidobacterium animalis, 10-30% of Paracaseicola casei, 10-30% of Lactobacillus plantarum, and 10-20% of Lactobacillus acidophilus.

3. The S. cerevisiae probiotic formulation of claim 1, wherein, Saccharomyces cerevisiae CNCM 1-3799, probiotic bacteria powder, blueberry fruit powder are all 40 mesh granular, the number of viable bacteria are respectively Saccharomyces cerevisiae CNCM 1-3799≥1×10 10 CFU / g, probiotic bacteria powder≥5×10 10 CFU / g.

4. The Saccharomyces cerevisiae probiotic formulation of claim 1, wherein, The culture medium raw material is 180-200 parts of glucose, 50-80 parts of yeast extract, 10-15 parts of galacto-oligosaccharides, 8-12 parts of maltoligosaccharides, 1-1.5 parts of potassium citrate chelated zinc, 5-8 parts of wheat protein peptides, 0.001-0.002 parts of vitamin B 12 .

5. The S. cerevisiae probiotic formulation of claim 4, wherein, The preparation method of potassium citrate chelated zinc includes the following steps: B1. Weigh 180-220 parts of citric acid, add 800-1000 parts of purified water, place it on a constant-temperature magnetic stirrer, stir at 30-40℃ until completely dissolved at a stirring speed of 300-500 r / min to obtain a citric acid solution; slowly add 80-85 parts of zinc carbonate to the citric acid solution, continue to stir at 30-35℃ for 20-30 min until no bubbles are generated to obtain a transparent zinc citrate solution; B2. Add 160-180 parts of potassium citrate to the zinc citrate solution, heat to 45-50℃, adjust the stirring speed to 500-800 r / min, adjust the pH to 5.5-6.0, and continuously react for 2-3 h; B3. Transfer the reaction solution into a vacuum rotary evaporator, set the temperature to 60-80℃ and the vacuum degree to 0.06-0.08 MPa, and concentrate to 1 / 5 of the original volume to obtain a concentrated solution; B4. Pour the concentrated solution into a freeze-drying tray, place it into a freeze-drying machine, pre-freeze at -20 to -40℃ for 2-5 h, maintain the vacuum degree at 5-10 Pa and the sublimation temperature at 30-40℃, dry for 8-12 h, pulverize by a pulverizer at a speed of 1000 r / min for 3 min, and sieve through a 40-mesh sieve to obtain potassium citrate chelated zinc.

6. The S. cerevisiae probiotic formulation of claim 1, wherein, The parameters of the activation tank in step A2 are rotation speed of 80-100 r / min, temperature of 30-32 DEG C, and ventilation of 0.5-1 L / (L·min); the parameters of the activation tank in step A3 are rotation speed of 80-100 r / min, temperature of 30-35 DEG C, and ventilation of 0.8-1.2 L / (L·min).

7. The method for preparing the Saccharomyces cerevisiae probiotic preparation according to any one of claims 1 to 6, characterized in that, The method comprises the following steps: S1, grinding and sieving the Saccharomyces cerevisiae CNCMI-3799, probiotic bacteria powder and blueberry fruit powder to 40 mesh standard; S2, adding fructooligosaccharide, malt dextrin, xylooligosaccharide, erythritol, blueberry fruit powder, and activated Saccharomyces cerevisiae CNCMI-3799 and probiotic bacteria powder into a mixer to stir to obtain a mixture, and setting the mixing time to 20-30 min and the mixing speed to 20-30 r / min; S3, selecting a vertical fluidized bed granulator, and feeding the mixture into the fluidized bed of the fluidized bed granulator, starting the air inlet system, and after the temperature in the bed is stabilized to the set range, starting the atomizing nozzle to spray purified water, and when the particle size preliminarily reaches 30-40 mesh, stopping the liquid spraying and continuing to ventilate for 3 min; S4, placing the granulated material into a hot air drying oven, setting the drying temperature to 50-60 DEG C, and setting the drying time to 1-2 h, sieving the granules through a 40 mesh sieve, feeding the sieved granules into a horizontal mixer, adding an anti-caking agent at 0.4-0.6% of the total weight of the granules, setting the rotation speed to 80-100 r / min, and mixing for 5-8 min, and after the mixing is completed, packaging is performed using a full-automatic strip-shaped bag packaging machine, and the parameter settings are as follows: the bag type is a three-edge sealed strip-shaped bag, the specification is 10 cm x 6 cm, the heat sealing temperature is 120-130 DEG C, the heat sealing time is 0.8 s, the filling speed is 40-50 bags / min, and the net content of each bag is 3 g.

8. The method of claim 7, wherein the Saccharomyces cerevisiae probiotic preparation is prepared by, The process parameters of the vertical fluidized bed granulator in step S3 are set as follows: the air inlet temperature is 60-65 DEG C, the air outlet temperature is 35-40 DEG C, the atomizing pressure is 0.22-0.25 MPa, the liquid spraying speed is 6-8 mL / min, and the fluidized bed air speed is 1.2-1.5 m / s.

9. The method of claim 7, wherein the Saccharomyces cerevisiae probiotic preparation is prepared by, In step S4, the anti-caking agent is one of fumed silica, magnesium stearate, and tricalcium phosphate; the fumed silica has a particle size of 5-10 μm and a purity of ≥99.8%; the magnesium stearate has a particle size of 10-20 μm and a purity of ≥99%; and the tricalcium phosphate has a particle size of 20-30 μm, a calcium content of ≥38%, and a phosphorus content of ≥18%.

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