A method for the preparation of a probiotic spray-dried protectant, and a method for spray-drying probiotics

Abstract

The application discloses a preparation method of probiotic spray drying protective agent and a probiotic spray drying method. The preparation method of the probiotic spray drying protective agent comprises the following steps: homogenizing rice wine residue to obtain a homogenate; performing wall breaking treatment on the homogenate and then performing extraction to obtain rice wine residue extract; performing sterilization on the rice wine residue extract and then performing centrifugation to collect rice wine residue precipitate; performing wall breaking and beating of the rice wine residue precipitate, adding beta-cyclodextrin into the rice wine residue precipitate, and then performing filtration and collecting a filtrate, so that the probiotic spray drying protective agent is prepared. The probiotic spray drying protective agent is prepared by mixing beta-cyclodextrin with rice wine residue after homogenization, wall breaking, water bath extraction, sterilization and centrifugation, and can protect probiotic bacteria when the probiotic bacteria are subjected to spray drying, effectively protects the bacteria, improves the survival rate of the bacteria, is resistant to gastric juice, has a high intestinal release rate, and reduces the raw material cost of the probiotic spray drying.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of probiotic preparation, in particular to a preparation method of probiotic spray drying protective agent and a probiotic spray drying method. BACKGROUND

[0002] Probiotics need to use high temperature in the process of spray drying, and the strains will face the effects of dehydration, osmotic pressure, high temperature, oxygen stimulation, which is very unfavorable to the survival rate of probiotics. According to the existing research, the protective agent added in spray drying is generally protein, lipid or sugar, which has an unsatisfactory effect and high cost, and it is necessary to reduce the raw material cost of probiotic spray drying. SUMMARY

[0003] The main purpose of the present application is to provide a preparation method of probiotic spray drying protective agent and a probiotic spray drying method, which aims to provide a low-cost and effective probiotic spray drying protective agent.

[0004] To achieve the above purpose, the present application provides a preparation method of probiotic spray drying protective agent, which comprises the following steps:

[0005] The rice wine residue is homogenized to obtain a homogenate;

[0006] The homogenate is subjected to wall breaking treatment and then extracted to obtain a rice wine residue extract;

[0007] The rice wine residue extract is sterilized and then centrifuged to collect rice wine residue precipitate;

[0008] The rice wine residue precipitate is subjected to wall breaking and pulping, and β-cyclodextrin is added thereto, and then filtered and the filtrate is collected to obtain the probiotic spray drying protective agent.

[0009] Optionally, the step of homogenizing the rice wine residue to obtain a homogenate comprises:

[0010] The rice wine residue is filtered and then put into a tissue crushing homogenizer, and homogenized at a speed of 10000-14000r / min for 3-8min.

[0011] Optionally, the step of subjecting the homogenate to wall breaking treatment and then extracting to obtain a rice wine residue extract comprises:

[0012] The homogenate is subjected to wall breaking treatment at an ultrasonic power of 100-300W for 15-20min, and then heated in water bath at 70-80℃ for 30-35min to obtain the rice wine residue extract.

[0013] Optionally, the step of sterilizing the rice wine residue extract and then centrifuging to collect rice wine residue precipitate comprises:

[0014] After mixing the rice wine lees with ultrapure water, sterilize at 120-125℃ for 15-20min, then centrifuge and collect the rice wine lees precipitate.

[0015] Optionally, the step of preparing the probiotic spray-drying protective agent by breaking the cell wall of the rice wine lees precipitate and adding β-cyclodextrin to it, then filtering and collecting the filtrate, comprises:

[0016] Put the rice wine lees precipitate into a cell wall breaker, add water to it, then break the cell wall, add β-cyclodextrin, mix uniformly, then filter and collect the filtrate to prepare the probiotic spray-drying protective agent; wherein 5-25g of the rice wine lees precipitate is added to 100mL of water.

[0017] Optionally, in the step of preparing the probiotic spray-drying protective agent by breaking the cell wall of the rice wine lees precipitate and adding β-cyclodextrin to it, then filtering and collecting the filtrate:

[0018] The mass ratio of the rice wine lees precipitate to the β-cyclodextrin is 5-25:15.

[0019] Further, the present application also proposes a method for probiotic spray-drying, comprising the following steps:

[0020] Mix the probiotic bacterial suspension to be spray-dried with the probiotic spray-drying protective agent, then place it in a spray dryer for spray-drying; wherein the probiotic spray-drying protective agent is prepared by the method for preparing the probiotic spray-drying protective agent as described above.

[0021] Optionally, the probiotics comprise at least one of Lactobacillus plantarum, Lactobacillus reuteri, Bifidobacterium lactis and Paracasei.

[0022] Optionally, in the step of mixing the probiotic bacterial suspension to be spray-dried with the probiotic spray-drying protective agent, then placing it in a spray dryer for spray-drying:

[0023] 5-25g of the probiotic spray-drying protective agent is added to 8mL of the probiotic bacterial suspension.

[0024] Optionally, in the step of mixing the probiotic bacterial suspension to be spray-dried with the probiotic spray-drying protective agent, then placing it in a spray dryer for spray-drying:

[0025] The parameters of the spray dryer are set as follows: needle frequency 1-2s, needle pressure 0.3-0.5MPa, pressure 0.1-0.2MPa, air inlet temperature 100-120℃, feeding speed 30-50rpm, and fan frequency 30-50Hz.

[0026] The technical scheme provided by the present application is to use rice wine residue as raw material, and after uniform homogenization, wall breaking, water bath extraction, sterilization, centrifugation and other treatments, the rice wine residue is mixed with beta-cyclodextrin to form a probiotic spray drying protective agent. The rice wine residue contains a large amount of small and large molecule proteins, polypeptides, starch, polysaccharides, disaccharides and other substances. The types of the contained components are various, and can interact and synergize. The probiotic spray drying protective agent can protect the probiotic bacteria during spray drying, and the disaccharides can replace the water removed during spray drying to protect the cell membrane structure. The proteins and polysaccharides can form a wall material to effectively protect the bacterial body and improve the survival rate of the bacterial strain. In addition, the probiotic spray drying protective agent uses rice wine residue as raw material, which belongs to waste resource recycling, greatly reducing the production cost of the probiotic spray drying protective agent, thereby significantly reducing the raw material cost of the probiotic spray drying. BRIEF DESCRIPTION OF DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other related drawings can also be obtained without creative labor.

[0028] Figure 1 The flowchart of an embodiment of the preparation method of the probiotic spray drying protective agent provided by the present application.

[0029] The implementation, functional characteristics and advantages of the present application will be further described with reference to the embodiments and the accompanying drawings. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and advantages of the embodiments of the present application more clear, the technical scheme in the embodiments of the present application will be described clearly and completely. The specific conditions not mentioned in the embodiments are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments not mentioned by the manufacturer are conventional products that can be purchased in the market. In addition, the meaning of "and / or" in the full text includes three parallel schemes. Taking "A and / or B" as an example, it includes A scheme, or B scheme, or A and B scheme. In addition, the technical schemes of each embodiment can be combined with each other, but it must be based on the fact that a person skilled in the art can realize it. When the combination of technical schemes appears contradictory or unachievable, it should be considered that the combination of technical schemes does not exist and is not within the scope of protection required by the present application. Based on the embodiments in the present application, all other embodiments obtained by a person skilled in the art without creative labor are within the scope of protection of the present application.

[0031] The probiotic bacteria need to use high temperature in the process of spray drying, and the strains will face the effects of dehydration, osmotic pressure, high temperature, oxygen stimulation, which is very unfavorable to the survival rate of probiotic bacteria. According to the existing research, the protective agent added in spray drying is generally protein, lipid or sugar, but the protective effect on probiotic bacteria is not ideal, and the survival rate of probiotic bacteria needs to be further improved.

[0032] Therefore, the application provides a preparation method of a probiotic spray drying protective agent, which is prepared from rice wine lees, can effectively improve the strain survival rate of probiotic bacteria, and an embodiment of the preparation method of the probiotic spray drying protective agent is shown in the figure. Figure 1 As shown in the figure, in the embodiment, the preparation method of the probiotic spray drying protective agent comprises the following steps:

[0033] Step S10, homogenate treatment is performed on the rice wine lees to obtain a homogenate liquid;

[0034] Step S20, after the homogenate liquid is subjected to cell wall breaking treatment and then leaching, a rice wine lees leaching liquid is obtained;

[0035] Step S30, after the rice wine lees leaching liquid is sterilized and then centrifuged, a rice wine lees precipitate is collected;

[0036] Step S40, the rice wine lees precipitate is subjected to cell wall breaking and pulping, and then β-cyclodextrin is added, and then the filtrate is collected by filtration, so that the probiotic spray drying protective agent is prepared.

[0037] Firstly, the rice wine lees is subjected to homogenate treatment to obtain a homogenate liquid. The rice wine lees can be directly obtained as the residue after rice wine brewing (usually waste in the rice wine production process), or can be obtained by brewing with glutinous rice as raw material. The specific brewing process can refer to the prior art, for example, it can be: glutinous rice→soaking→water filtering→washing→steaming rice→cooling→mixing koji and water→filling jar→fermentation→pressing→clarification→rice wine. Finally, the brewed rice wine is filtered to obtain the rice wine lees. Further, in some embodiments of the application, the homogenate treatment is performed by placing the rice wine lees in a homogenizer and homogenizing at a speed of 10000-14000 r / min for 3-8 min to obtain the homogenate liquid.

[0038] Then, the homogenate liquid is subjected to cell wall breaking treatment, and then leaching to obtain a rice wine lees leaching liquid. In some embodiments of the application, the cell wall breaking treatment is performed by treating the homogenate liquid under an ultrasonic power of 100-300 W for 15-20 min, and the leaching is performed by water bath heating at 70-80℃ for 30-35 min.

[0039] Then, the rice wine lees extract solution is added with ultrapure water, and after being mixed uniformly, sterilized and centrifuged, the rice wine lees precipitate is collected. In some embodiments of the present application, the sterilization is high-temperature sterilization, specifically, sterilization at 120-125 DEG C for 15-20 min; the centrifugation is performed at a speed of 4000-6000 r / min for 10-15 min.

[0040] The rice wine lees is the residue after fermentation of glutinous rice, and still retains the granular shape of glutinous rice. It is rich in nutrients, containing various sugars, B vitamins, minerals, various amino acids, and also has the functions of invigorating the spleen and stomach, relaxing the meridians and relieving pain and removing blood stasis, etc. It also contains carbon sources, nitrogen sources, inorganic salts and growth factors required for microbial growth. Therefore, the supernatant after centrifugation in step S30 can be used for proliferation culture of probiotic bacterial strains, and the rice wine lees precipitate after centrifugation is used for preparing the probiotic spray-drying protective agent.

[0041] Finally, the rice wine lees precipitate is broken and pulped, and then the obtained slurry is mixed with β-cyclodextrin, and then filtered with three layers of gauze to prevent the prepared probiotic spray-drying protective agent from blocking the spray nozzle during spray drying. The filtrate is collected after filtration, and the probiotic spray-drying protective agent is prepared. Specifically, in some embodiments of the present application, the broken and pulping is that the rice wine lees precipitate is put into a broken wall machine, an appropriate amount of water is added, and then pulping is performed. The amount of water added can be set to 5-25 g of the rice wine lees precipitate per 100 mL of water; the mass ratio of the β-cyclodextrin to the rice wine lees precipitate is 15:5-25.

[0042] In the technical solution provided by the present application, the rice wine lees is used as the raw material, and after uniform pulping, broken wall, water bath extraction, sterilization, centrifugation and other treatments, it is mixed with β-cyclodextrin to prepare a probiotic spray-drying protective agent. Since the rice wine lees contains rich macromolecular and small molecular proteins, polypeptides, starch, polysaccharides, disaccharides and other substances, the types of the contained components are many, and can play an interactive synergistic role. It can protect the probiotic bacterial cells during spray drying, the disaccharides can replace the water removed during spray drying to protect the structure of the cell membrane, and the proteins and polysaccharides can form a wall material to effectively protect the bacterial cells and improve the survival rate of the bacterial strains.

[0043] In addition, the utilization rate of rice wine residue is low, and it is wasted, and its value has not been effectively tapped, the annual output of rice wine is about 3.1 billion liters, and the output of rice wine residue is about 1 million tons, which is considered to be high-polluting, because its emission can cause negative effects on the ecosystem, and it is a great waste of resources. Therefore, the technical scheme of the present application uses rice wine residue as raw material to prepare probiotic spray drying protective agent, and also realizes resource recycling, fully utilizes rice wine residue resources, is conducive to reducing the pollution of rice wine residue emission to the environment, and compared with using skimmed milk powder as a protective agent, the price of skimmed milk powder is expensive, only 100g needs about 80 yuan, and the protective agent prepared by the method of the present application can greatly reduce the raw material cost of probiotic spray drying, and the rice wine residue itself is edible, and its addition will not affect the probiotic preparation.

[0044] In addition, the inventors also found during the research process that the probiotic spray drying protective agent prepared by using rice wine residue as raw material in the present application has the characteristics of good resistance to gastric juice and intestinal release rate when used as a protective agent in probiotic spray drying.

[0045] Based on the method provided in the present application, the probiotic spray drying protective agent prepared by the method has the characteristics of good resistance to gastric juice and intestinal release rate when used as a protective agent in probiotic spray drying.

[0046] The probiotic bacterial suspension to be spray dried is mixed with the probiotic spray drying protective agent, and then placed in a spray dryer for spray drying; wherein the probiotic spray drying protective agent is prepared by the preparation method of the probiotic spray drying protective agent as described above.

[0047] When the probiotic spray drying protective agent provided by the present application is used for spray drying of probiotic bacterial suspension, it can effectively protect the probiotic bacterial strain and improve the survival rate of the strain; at the same time, the probiotic powder prepared after spray drying also has the characteristics of good resistance to gastric juice and intestinal release rate.

[0048] The probiotic spray drying method provided by the present application can be applied to probiotics including but not limited to at least one of lactobacillus plantarum, lactobacillus reuteri, bifidobacterium lactis and paracasei lactobacillus, that is, the probiotic spray drying method provided by the present application can be used for spray drying treatment of any one of the above-mentioned probiotics, or can be used for spray drying treatment of mixed probiotics of two or more kinds, which all belong to the protection scope of the present application.

[0049] In addition, in some embodiments of the present application, the method for spray drying probiotics can further include a step of preparing a probiotic bacterial suspension, specifically, pouring the live bacteria powder in the probiotic capsules into 500 mL of sterilized normal saline, shaking and mixing for 10 min to obtain a probiotic liquid, and then sucking the probiotic liquid into MRS liquid medium and culturing for 2-3 generations to obtain the probiotic bacterial suspension, which can be used for subsequent spray drying treatment. In other embodiments of the present application, the probiotic bacterial suspension can also be prepared by other conventional methods in the art, which will not be described here.

[0050] Further, when the probiotic bacterial suspension to be spray dried is mixed with the probiotic spray drying protective agent, experiments have shown that the preferred mixing ratio is 5-25 g of the probiotic spray drying protective agent per 8 mL of the probiotic bacterial suspension. Within this range, the probiotic spray drying protective agent can effectively protect the probiotic bacterial strain under the high temperature, oxygen stimulation and other adverse environments existing in the spray drying process, while also avoiding the problem of dilution of the probiotic bacterial strain due to the clogging of the nozzle of the spray dryer by the protective agent, thereby reducing the survival rate of the strain.

[0051] Further, in some embodiments of the present application, the parameters of the spray dryer are set as follows: needle frequency 1-2 s, needle pressure 0.3-0.5 MPa, pressure 0.1-0.2 MPa, air inlet temperature 100-120℃, feeding speed 30-50 rpm, and fan frequency 30-50 Hz.

[0052] The technical solutions of the present application will be further described in detail below in combination with specific embodiments and drawings. It should be understood that the following embodiments are only used to explain the present application and do not limit the present application.

[0053] Example 1 Preparation of probiotic spray drying protective agent

[0054] (1) Brewing old rice wine: glutinous rice → soaking → filtering water → washing → steaming rice → spreading and cooling → mixing with koji and adding water → filling jars → fermentation → pressing → clarification → rice wine → filtration → rice wine residue.

[0055] (2) Preparation of rice wine residue extract: put the rice wine residue into a tissue homogenizer, homogenize at a speed of 10000 r / min for 8 min to obtain a homogenate; then transfer the obtained homogenate to a beaker, break the cell wall under an ultrasonic power of 100 W for 20 min, and then heat extract at 70℃ for 35 min to obtain the rice wine residue extract.

[0056] (3) Preparation of rice wine dreg precipitate: a certain amount of ultrapure water was added to the rice wine dreg extract, and after mixing evenly, it was sterilized at 120°C for 15 min. After sterilization, it was centrifuged at a speed of 4000 r / min for 15 min, and the rice wine dreg precipitate was collected.

[0057] (4) 5 g of rice wine dreg precipitate was put into a cell disruptor, 100 mL of water was added, and the slurry was beaten. Then 15 g of β-cyclodextrin was added to the slurry, and the filtrate was collected after filtering with three layers of gauze to prepare a probiotic spray drying protective agent.

[0058] Example 2 Preparation of probiotic spray drying protective agent

[0059] (1) Brewing of old rice wine: glutinous rice → soaking → filtering water → washing with water → steaming rice → spreading and cooling → mixing with koji and adding water → filling jar → fermentation → pressing → clarification → rice wine → filtering → rice wine dregs.

[0060] (2) Preparation of rice wine dreg extract: the rice wine dregs were put into a tissue crushing homogenizer and homogenized at a speed of 11000 r / min for 6 min to obtain a homogenate. Then the obtained homogenate was transferred to a beaker and treated with ultrasonic wave at a power of 150 W for 18 min, followed by heating in water bath at 75°C for 32 min to obtain the rice wine dreg extract.

[0061] (3) Preparation of rice wine dreg precipitate: a certain amount of ultrapure water was added to the rice wine dreg extract, and after mixing evenly, it was sterilized at 120°C for 15 min. After sterilization, it was centrifuged at a speed of 4000 r / min for 15 min, and the rice wine dreg precipitate was collected.

[0062] (4) 5 g of rice wine dreg precipitate was put into a cell disruptor, 100 mL of water was added, and the slurry was beaten. Then 15 g of β-cyclodextrin was added to the slurry, and the filtrate was collected after filtering with three layers of gauze to prepare a probiotic spray drying protective agent.

[0063] Example 2 Preparation of probiotic spray drying protective agent

[0064] (1) Brewing of old rice wine: glutinous rice → soaking → filtering water → washing with water → steaming rice → spreading and cooling → mixing with koji and adding water → filling jar → fermentation → pressing → clarification → rice wine → filtering → rice wine dregs.

[0065] (2) Preparation of rice wine dreg extract: the rice wine dregs were put into a tissue crushing homogenizer and homogenized at a speed of 11000 r / min for 6 min to obtain a homogenate. Then the obtained homogenate was transferred to a beaker and treated with ultrasonic wave at a power of 150 W for 18 min, followed by heating in water bath at 75°C for 32 min to obtain the rice wine dreg extract.

[0066] (3) Preparation of rice wine dreg precipitate: a certain amount of ultrapure water was added to the rice wine dreg extract, and after mixing evenly, it was sterilized at 120°C for 15 min. After sterilization, it was centrifuged at a speed of 5000 r / min for 14 min, and the rice wine dreg precipitate was collected.

[0067] (4) 15 g of rice wine dreg precipitate was put into a cell disruptor, 100 mL of water was added, and the slurry was beaten. Then 15 g of β-cyclodextrin was added to the slurry, and the filtrate was collected after filtering with three layers of gauze to prepare a probiotic spray drying protective agent.

[0068] Example 4 Preparation of probiotic spray drying protective agent

[0069] (1) Brewing of old rice wine: glutinous rice → soaking → filtering water → washing with water → steaming rice → spreading and cooling → mixing with koji and adding water → filling jar → fermentation → pressing → clarification → rice wine → filtering → rice wine dregs.

[0070] (2) Preparation of rice wine dreg extract: the rice wine dregs were put into a tissue crushing homogenizer and homogenized at a speed of 13000 r / min for 4 min to obtain a homogenate. Then the obtained homogenate was transferred to a beaker and treated with ultrasonic wave at a power of 250 W for 16 min, followed by heating in water bath at 78°C for 31 min to obtain a rice wine dreg extract.

[0071] (3) Preparation of rice wine dreg precipitate: a certain amount of ultrapure water was added to the rice wine dreg extract, and after mixing evenly, it was sterilized at 120°C for 15 min. After sterilization, it was centrifuged at a speed of 5000 r / min for 14 min, and the rice wine dreg precipitate was collected.

[0072] (4) 15 g of rice wine dreg precipitate was put into a cell disruptor, 100 mL of water was added, and the slurry was beaten. Then 15 g of β-cyclodextrin was added to the slurry, and the filtrate was collected after filtering with three layers of gauze to prepare a probiotic spray drying protective agent.

[0073] Example 5 Preparation of probiotic spray drying protective agent

[0074] (1) Brewing of old rice wine: glutinous rice → soaking → filtering water → washing with water → steaming rice → spreading and cooling → mixing with koji and adding water → filling jar → fermentation → pressing → clarification → rice wine → filtering → rice wine dregs.

[0075] (2) Preparation of rice wine dreg extract: the rice wine dregs were put into a tissue crushing homogenizer and homogenized at a speed of 13000 r / min for 4 min to obtain a homogenate. Then the obtained homogenate was transferred to a beaker and treated with ultrasonic wave at a power of 250 W for 16 min, followed by heating in water bath at 78°C for 31 min to obtain a rice wine dreg extract.

[0076] (3) Preparation of rice wine residue precipitate: a certain amount of ultrapure water was added to the rice wine residue extraction solution, which was mixed uniformly and sterilized at 120°C for 20 min. After sterilization, the rice wine residue precipitate was collected by centrifugation at a speed of 6000 r / min for 10 min.

[0077] (4) 25 g of rice wine residue precipitate was placed in a cell disrupter, 100 mL of water was added, and the slurry was beaten. Then 15 g of β-cyclodextrin was added to the slurry, which was filtered with three layers of gauze to collect the filtrate, thereby preparing a probiotic spray-drying protective agent.

[0078] Example 6 Method for spray-drying probiotics

[0079] (1) Preparation of probiotic bacterial suspension: the live bacteria powder in the probiotic capsule was poured into 500 mL of sterilized normal saline, shaken for 10 min, and then the probiotic liquid was obtained. The probiotic liquid was then taken into MRS liquid medium and cultured for 2-3 generations to obtain a probiotic bacterial suspension.

[0080] (2) The probiotic bacterial suspension prepared in step (1) was mixed with the probiotic spray-drying protective agent prepared in Example 1 according to the ratio of 5 g of probiotic spray-drying protective agent per 8 mL of probiotic bacterial suspension, and then stirred uniformly. The rubber tube was connected to the spray dryer, and the parameters of the spray dryer were adjusted to spray-dry the probiotic bacterial suspension to obtain a probiotic powder. The parameters of the spray dryer were set as follows: needle frequency 1 s, needle pressure 0.4 MPa, pressure 0.1 MPa, inlet air temperature 100°C, feeding speed 30 rpm, and fan frequency 30 Hz.

[0081] Example 7 Method for spray-drying probiotics

[0082] (1) Preparation of probiotic bacterial suspension: the live bacteria powder in the probiotic capsule was poured into 500 mL of sterilized normal saline, shaken for 10 min, and then the probiotic liquid was obtained. The probiotic liquid was then taken into MRS liquid medium and cultured for 2-3 generations to obtain a probiotic bacterial suspension.

[0083] (2) The probiotic bacterial suspension prepared in step (1) was mixed with the probiotic spray-drying protective agent prepared in Example 2 according to the ratio of 10 g of probiotic spray-drying protective agent per 8 mL of probiotic bacterial suspension, and then stirred uniformly. The rubber tube was connected to the spray dryer, and the parameters of the spray dryer were adjusted to spray-dry the probiotic bacterial suspension to obtain a probiotic powder. The parameters of the spray dryer were set as follows: needle frequency 1-2 s, needle pressure 0.5 MPa, pressure 0.2 MPa, inlet air temperature 105°C, feeding speed 35 rpm, and fan frequency 35 Hz.

[0084] Example 8 Method for spray-drying probiotics

[0085] (1) Preparation of probiotic bacterial suspension: pour the live bacteria powder in the probiotic capsule into 500 mL of sterilized normal saline, shake for 10 min, obtain the probiotic bacterial liquid, then take the probiotic bacterial liquid into MRS liquid medium, culture for 2-3 generations, obtain the probiotic bacterial suspension.

[0086] (2) According to the proportion of 15 g of probiotic spray drying protective agent corresponding to 8 mL of probiotic bacterial suspension, mix and stir the probiotic bacterial suspension prepared in step (1) and the probiotic spray drying protective agent prepared in Example 3, connect to the spray dryer with a rubber tube, adjust the parameters of the spray dryer, and then spray dry the probiotic bacterial suspension to obtain the probiotic powder; wherein the parameters of the spray dryer are set as: needle frequency 1 s, needle pressure 0.3 MPa, pressure 0.1 MPa, inlet air temperature 110°C, feeding speed 40 rpm, and fan frequency 40 Hz.

[0087] Example 9 Method for spray drying of probiotics

[0088] (1) Preparation of probiotic bacterial suspension: pour the live bacteria powder in the probiotic capsule into 500 mL of sterilized normal saline, shake for 10 min, obtain the probiotic bacterial liquid, then take the probiotic bacterial liquid into MRS liquid medium, culture for 2-3 generations, obtain the probiotic bacterial suspension.

[0089] (2) According to the proportion of 15 g of probiotic spray drying protective agent corresponding to 8 mL of probiotic bacterial suspension, mix and stir the probiotic bacterial suspension prepared in step (1) and the probiotic spray drying protective agent prepared in Example 3, connect to the spray dryer with a rubber tube, adjust the parameters of the spray dryer, and then spray dry the probiotic bacterial suspension to obtain the probiotic powder; wherein the parameters of the spray dryer are set as: needle frequency 1 s, needle pressure 0.3 MPa, pressure 0.1 MPa, inlet air temperature 110°C, feeding speed 40 rpm, and fan frequency 40 Hz.

[0090] Example 10 Method for spray drying of probiotics

[0091] (1) Preparation of probiotic bacterial suspension: pour the live bacteria powder in the probiotic capsule into 500 mL of sterilized normal saline, shake for 10 min, obtain the probiotic bacterial liquid, then take the probiotic bacterial liquid into MRS liquid medium, culture for 2-3 generations, obtain the probiotic bacterial suspension.

[0092] (2) According to the proportion of 25 g of probiotic spray-drying protective agent added for every 8 mL of probiotic bacterial suspension, the probiotic bacterial suspension prepared in step (1) is mixed with the probiotic spray-drying protective agent prepared in Example 5 to be stirred uniformly, connected to a spray dryer by a rubber tube, and subjected to spray drying treatment after adjusting the parameters of the spray dryer, to obtain probiotic powder; wherein the parameters of the spray dryer are set as follows: needle frequency 1 s, needle pressure 0.5 MPa, pressure 0.1 MPa, inlet air temperature 120℃, feeding speed 50 rpm, and fan frequency 50 Hz.

[0093] Comparative Example 1

[0094] The steps are the same as those in Example 6, except that no probiotic spray-drying protective agent is added in step (2).

[0095] The viable counts of the probiotic powders subjected to spray drying in Examples 6 to 10 and Comparative Example 1 are detected, and the detection method is as follows: 1 g of the reconstituted spray-dried sample is added to 9 ml of sterilized normal saline, mixed uniformly by a vortex oscillator, subjected to 10-fold serial dilution, and the sample liquid of a suitable dilution is selected, 1 ml of which is taken and added to a sterile glass flat plate, poured into sterilized MRS culture medium, and placed in a 37℃ incubator for culture. The viable count is detected, and three sets of parallel samples are prepared for each gradient, and the average value is taken.

[0096] According to the detection results of the viable counts of the probiotic powders, the strain survival rates are calculated, and the results are shown in Table 1.

[0097] Table 1 Strain survival rates of probiotic powders subjected to spray drying in examples and comparative examples

[0098] Comparative Example 1 Example 6 Example 7 Example 8 Example 9 Example 10 Survival rate / % 3.31 6.75 12.9 28.5 15.29 13.66

[0099] As can be seen from the data in Table 1, after the probiotic spray-drying protective agent prepared from rice wine lees is added in the examples of the present application, the strain survival rate of the probiotic powder obtained after spray drying is significantly improved, and within a certain range, the protection of the strain during spray drying is better and the survival rate gradually increases as the amount of the protective agent increases, but when the amount of the protective agent is too large, the survival rate decreases to a certain extent, which may be because as the amount of the probiotic powder increases, the dried probiotic powder is equivalent to being diluted, and thus the survival rate of the strain may decrease.

[0100] The probiotic powders subjected to spray drying in the examples are subjected to gastrointestinal stress and release simulation, and simulation gastric juice test is carried out by taking Examples 6, 8 and 10 as examples, and simulation small intestinal juice test is carried out by taking Example 8 as an example, and the test methods and results are as follows:

[0101] 0.1 g of the spray-dried probiotic powder was added to 0.9 g of phosphate buffer with a concentration of 0.2 M (pH = 7.0) and allowed to swell for 10 min, and then the mixed solution was added to 9 g of preheated gastric juice or artificial small intestinal juice and incubated in a water bath shaker at 37 °C and 100 rpm for 0-150 min. The sample treated by gastric juice or artificial small intestinal juice was centrifuged at 0-4 °C and 5000-6000 rpm for 5-10 min to obtain the spray-dried probiotic powder precipitate and re-dispersed into 9.9 g of phosphate buffer. For the simulated gastric juice sample, the pH of the mixture should be adjusted to 6.0-8.0 before centrifugation. The survival cells after gastric juice or intestinal juice treatment were detected by plate counting according to the method for detecting viable cells described above. The spray-dried probiotic powder treated with normal saline was used as a control for the survival of bacteria in simulated gastric and intestinal juice.

[0102] The results of detecting the viable cell count of the strain after simulated gastric juice digestion are shown in Table 2, and the results of detecting the viable cell count of the strain after simulated small intestinal juice digestion are shown in Table 3.

[0103] Table 2 Viable cell count after simulated gastric juice digestion

[0104]

[0105] Table 3 Viable cell count after simulated small intestinal juice digestion

[0106]

[0107]

[0108] As can be seen from the data in Table 2, the survival rates of the control and the examples in the simulated gastric juice digestion are different due to the different proportions of the spray-dried probiotic protection agent prepared in the examples of the present application added. It can be seen that the strain in the control group has no survival in the gastric juice, while the spray-dried probiotic protection agent provided by the present application has a certain protective effect on the bacterial cells, and the more the protective agent is added, the greater the tolerance of the probiotic powder to the gastric juice.

[0109] As can be seen from the data in Table 3, in the simulated small intestinal juice, the spray-dried sample slowly decreased by 0.45 lg CFU / g, indicating that the spray-dried sample has strong tolerance to bile salts and trypsin. The spray-dried probiotic powder prepared in the examples of the present application can better resist the high osmotic pressure of bile salts in the intestinal tract.

[0110] The above are only preferred embodiments of the present application, and do not limit the patent scope of the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the patent protection scope of the present application.

Claims

1. A method of preparing a probiotic spray-dried protectant, characterized in that, The method comprises the following steps: homogenate the rice wine lees to obtain a homogenate liquid; subject the homogenate liquid to wall breaking treatment and then extract to obtain a rice wine lees extract liquid; sterilize the rice wine lees extract liquid and then centrifuge to collect rice wine lees precipitate; put the rice wine lees precipitate into a wall breaking machine, add water to the wall breaking machine, then beat the slurry, add β-cyclodextrin, mix uniformly, filter and collect the filtrate to obtain a probiotic spray drying protective agent; wherein 5-25 g of the rice wine lees precipitate is added to 100 mL of water, and the mass ratio of the rice wine lees precipitate to the β-cyclodextrin is 5-25:

15.

2. The method of claim 1, wherein the probiotic spray-dried protectant is prepared by, The step of homogenating the rice wine lees to obtain a homogenate liquid comprises: put the filtered rice wine lees into a tissue crushing homogenizer, and homogenate at a speed of 10,000-14,000 r / min for 3-8 min.

3. The method of claim 1, wherein the probiotic spray-dried protectant is prepared by, The step of subjecting the homogenate liquid to wall breaking treatment and then extracting to obtain a rice wine lees extract liquid comprises: subject the homogenate liquid to wall breaking treatment at an ultrasonic power of 100-300 W for 15-20 min, and then heat in a water bath at 70-80 ℃ for 30-35 min to extract to obtain the rice wine lees extract liquid.

4. The method of claim 1, wherein the probiotic spray-dried protectant is prepared by, The step of sterilizing the rice wine lees extract liquid and then centrifuging to collect rice wine lees precipitate comprises: mix the rice wine lees with ultrapure water, sterilize at 120-125 ℃ for 15-20 min, then centrifuge and collect the rice wine lees precipitate.

5. A method of spray drying probiotic bacteria, characterized in that, The method comprises the following steps: mix the probiotic bacterial suspension to be spray dried with the probiotic spray drying protective agent, and then place in a spray dryer for spray drying; wherein the probiotic spray drying protective agent is prepared by the method for preparing a probiotic spray drying protective agent according to any one of claims 1 to 4.

6. The method of spray-drying probiotic bacteria according to claim 5, wherein, The probiotic bacteria comprise at least one of Lactobacillus plantarum, Lactobacillus reuteri, Bifidobacterium lactis and Paracaseicobacter.

7. The method of spray-drying probiotic bacteria according to claim 5, wherein, In the step of mixing the probiotic bacterial suspension to be spray dried with the probiotic spray drying protective agent, and then placing in a spray dryer for spray drying: 5-25 g of the probiotic spray drying protective agent is added to 8 mL of the probiotic bacterial suspension.

8. The method of spray drying probiotic bacteria according to claim 5, wherein, In the step of mixing the probiotic bacterial suspension to be spray dried with the probiotic spray drying protective agent, and then placing in a spray dryer for spray drying: the parameters of the spray dryer are set as follows: needle frequency 1-2 s, needle pressure 0.3-0.5 MPa, pressure 0.1-0.2 MPa, air inlet temperature 100-120 ℃, feeding speed 30-50 rpm, and fan frequency 30-50 Hz.

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