CN112391303B - Bifidobacterium powder and preparation process of probiotics thereof - Google Patents

Bifidobacterium powder and preparation process of probiotics thereof Download PDF

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CN112391303B
CN112391303B CN201910751796.7A CN201910751796A CN112391303B CN 112391303 B CN112391303 B CN 112391303B CN 201910751796 A CN201910751796 A CN 201910751796A CN 112391303 B CN112391303 B CN 112391303B
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bifidobacterium
inulin
mannitol
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林楚雄
黄敏鑫
陈晓玲
卢君
刘英敏
李国伟
曾维芬
何彩霞
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Guangdong Changxing Biotechnology Co ltd
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    • A23V2400/51Bifidobacterium

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Abstract

The invention discloses a bifidobacteria powder and a preparation process of probiotics thereof, wherein a protective matrix containing maltose, calcium hydrophosphate and poloxamer is selected, and thalli are mixed with the matrix before spray drying, so that the heat resistance and stress resistance activity of bifidobacteria is improved, and the survival rate of the bifidobacteria in spray drying is obviously improved; not only can play a role in improving the survival rate of the bifidobacteria, but also can ensure that the dried bifidobacteria powder has good fluidity, thereby facilitating the subsequent preparation processing. The bifidobacterium powder prepared by the method is used as a raw material, a proper amount of milk powder, inulin, mannitol and silicon dioxide are added as auxiliary materials, and the powder packaged in a single dose is prepared by applying modern industrial technology.

Description

Bifidobacterium powder and preparation process of probiotics thereof
Technical Field
The invention relates to a preparation process of bifidobacterium powder and a preparation process of probiotics containing the bifidobacterium powder.
Background
Probiotics refers to a micro-ecological preparation which can maintain the balance of flora in human body and has beneficial effect on human health. The oral administration of sufficient active probiotics is helpful for relieving acute and chronic gastroenteritis, treating diarrhea, improving digestion, relieving symptoms such as lactose intolerance and the like. Bifidobacteria are a common probiotic, and the thallus preparation of the bifidobacteria is divided into a liquid preparation and a solid preparation. The former is prepared through traditional fermentation culture to obtain bifidobacterium liquid as the preparation product, and the latter is prepared through proliferation of probiotics, freeze drying, spray drying, embedding and other steps. The liquid preparation is easy to be polluted or the strain is degenerated along with the prolonging of the preservation time, the activation culture and the expansion culture are needed before the use, the solid microbial inoculum can be directly put into the culture, the expansion culture is not needed before the fermentation, and the preservation cost is lower. At present, the bifidobacterium solid preparation process is usually vacuum freeze drying, spray drying, fluidized bed drying, microwave vacuum drying and the like. Vacuum freeze drying has little damage to thallus subcells and high viable bacteria rate, but is easily influenced by factors such as pre-freezing temperature, freezing speed, vacuum degree and the like, and has long time consumption and high cost; the fluidized bed has longer drying time, and when the material stays for uneven time, the drying is uneven; in the microwave vacuum drying, polar molecules (such as thallus moisture) synchronously rotate at high speed along with microwave frequency, so that friction heat is instantaneously generated on materials, the surfaces and the interiors of the materials are simultaneously heated, a large number of thallus water molecules escape, and drying is realized. The spray drying is to directly spray the prepared bacterial suspension into mist through an atomizing device, and the mist is directly contacted with high-temperature air in a drying chamber to carry out heat and mass transfer, the drying time is extremely short, the material temperature is low, the product dispersibility and dissolubility are good, the process is simple, the equipment cost is relatively low, and the spray drying is particularly suitable for industrial continuous production and is widely used in the fields of medicines and foods.
However, the solution is fully mixed with high-temperature air in the spray drying process, so that the temperature of the bacteria liquid is sharply increased, and a large amount of bifidobacteria are killed. Therefore, a series of works have been made in the prior art on how to improve the survival rate of bifidobacteria. CN108070542A firstly preparing a bifidobacterium biomembrane, and then carrying out spray drying on the bifidobacterium biomembrane to ensure that the survival rate of the bifidobacterium spray-dried can reach more than 55 percent; CN105341940A, CN1306021C and CN102210659B disclose methods for improving the survival rate of bifidobacteria by optimizing the formulation of dry protective agent through screening of sugar, protein, salt, etc., or improving the survival rate through microencapsulation.
In addition, the bifidobacterium powder obtained in the spray drying process is usually processed into solid dosage forms such as tablets and the like for convenient oral administration. It is therefore also desirable for the powder obtained after spray drying to have good flowability (no caking ) and good formability (the tablets produced have a relatively high breaking force). In the prior art, an anti-caking agent and an adhesive are additionally added in the forming process, so that the subsequent preparation process is complicated.
In order to overcome the defects of the prior art, the process for developing the novel bifidobacterium powder has wide application prospect. Meanwhile, the bifidobacterium powder prepared by the method is used as a raw material, a proper amount of milk powder, inulin, mannitol and silicon dioxide are added as auxiliary materials, and the powder packaged in single dose is prepared by applying modern industrial technology.
Disclosure of Invention
The invention aims to provide a spray drying process of bifidobacterium powder, which has the advantages of high survival rate of bifidobacterium and good fluidity and formability of dried powder and solves the problems of low survival rate of the bifidobacterium after spray drying and complex subsequent preparation process in the prior art.
To achieve the above object, a first aspect of the present invention provides a method for producing a bifidobacterium powder, comprising:
a preparation method of bifidobacterium powder is characterized by comprising the following steps:
a. culturing Bifidobacterium in culture medium, and collecting thallus;
b. uniformly mixing the thalli and a spray drying matrix to obtain a thalli mixed solution;
c. spray drying, and collecting Bacillus bifidus powder;
wherein in step b the spray dried matrix comprises maltose, dibasic calcium phosphate and poloxamer.
In one embodiment, the medium of step a is TPY medium; the bifidobacterium strain is Chr, Hansen BB-12; the method comprises the following specific steps: inoculating Bifidobacterium into TPY culture medium at 5%, culturing at 37 + -3 deg.C for 16-24 hr under anaerobic condition, and culturing at 4 deg.C for 5000--1Centrifuging for 10-20min, discarding supernatant, cleaning thallus with sterilized distilled water, and centrifuging under the same condition to obtain Bifidobacterium thallus.
In one embodiment, the spray dried matrix of step b comprises: 7-13% by weight of skim milk, preferably 10% by weight of skim milk; 5-10% by weight of maltose, preferably 7-8% by weight; 1-2% by weight of calcium hydrogen phosphate, preferably 1.4-1.6% by weight; 0.3-0.5% by weight of poloxamer, preferably 0.35-0.45% by weight; the balance being water.
In a certain embodiment the poloxamer is selected from poloxamer 184, poloxamer 188 or poloxamer 407.
In one embodiment, the spray drying conditions of step c are: and c, drying the thallus mixed solution obtained in the step b by a spray dryer at the speed of 4-6ml/min, wherein the inlet temperature is 130-150 ℃, and the outlet temperature is 50-70 ℃. Spray drying to obtain Bifidobacterium powder.
The second aspect of the invention provides bifidobacterium powder prepared by the method.
The third aspect of the invention provides the application of the bacterium powder in the fields of food, health products and biology, and the application comprises the preparation of solid beverages, dairy products, fermented fruits and vegetables, chocolate, candies, microecologics, probiotic tablets, probiotic capsules and probiotic powder.
In one embodiment of the invention, the use comprises preparing a probiotic powder comprising bifidobacteria prepared according to the present invention, said probiotic powder further comprising lactobacillus acidophilus; preferably the probiotic powder further comprises a prebiotic component, for example comprising at least one of fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, breast milk oligosaccharides.
In one embodiment of the invention, the bifidobacterium powder is used for producing dairy products, bean products or fruit and vegetable products by fermentation.
In one embodiment of the invention, bifidobacterium powder prepared by the method is used as a raw material, a proper amount of milk powder, inulin, mannitol and silicon dioxide are added as auxiliary materials, and the probiotic powder packaged in a single dose is prepared by modern industrial technology.
The probiotic powder of the invention adopts milk powder, inulin, mannitol and silicon dioxide as auxiliary materials. Wherein, the milk powder: as a diluent of the product, the product also has the function of improving the mouthfeel; inulin: is a multiplication factor of bifidobacterium, and can greatly increase beneficial bacteria in colon and reduce pathogenic bacteria and putrefying bacteria by taking inulin every day; mannitol: the sweetener has the characteristics of no hygroscopicity, good chemical stability and the like, is used as a filling agent of the product, and also plays a role of a sweetener in the product due to certain sweetness; silicon dioxide: the product can be used as anticaking agent for improving powder flowability and preventing agglomeration.
The production process of the probiotic powder is simple, easy to operate and easy to control the product quality, and the main process route is as follows: weighing, mixing, subpackaging, packaging, obtaining finished products, inspecting and warehousing. Because the product has no terminal sterilization, except that the sanitary indexes of the raw and auxiliary materials and the packaged product are strictly controlled, the sanitary control (including production environment, equipment, containers and personnel) in the whole production process is strictly executed according to the GMP (good manufacturing practice) regulation of the health-care food, so as to ensure that the sanitary indexes of the product meet the quality requirements.
1) Preparation of raw materials: the bifidobacterium powder, the milk powder, the inulin, the mannitol and the silicon dioxide which are prepared by the method are strictly placed in a clean raw material workshop for standby after being subjected to outsourcing in a buffer room according to the procedure of entering a clean area of 10 ten thousand levels.
2) Pretreatment of raw materials and auxiliary materials: respectively sieving Bifidobacterium powder, milk powder, inulin, mannitol and silicon dioxide with 60 mesh sieve.
3) Weighing: according to the total formula amount and the formula proportion of each batch of ingredients, the bifidobacterium powder, the milk powder, the inulin, the mannitol and the silicon dioxide are accurately weighed, and the check is performed by a person skilled in the art.
4) Mixing: firstly, the bifidobacterium powder and the mannitol are gradually and uniformly mixed according to the equal quantity to obtain mixed powder B, and the mixed powder B, the milk powder, the inulin and the silicon dioxide are added into a three-dimensional mixer to be mixed for 25-35 minutes.
5) Subpackaging: subpackaging with a particle packaging machine according to requirements.
6) And (3) outer packaging: and (3) putting the subpackaged samples into a small box of 10 bags/box in an outer packaging room, labeling, boxing and packaging.
7) And (4) checking: and (5) inspecting the finished product according to the delivery inspection requirement of the product.
8) Warehousing: and (5) warehousing and storing the finished product after the finished product is qualified.
The equivalent incremental operation method comprises the steps of firstly taking a small amount of components, adding an equivalent large amount of components, then taking a large amount of components which are equivalent to the mixture, uniformly mixing, increasing by the times until all the large amount of components are added, uniformly mixing, and sieving.
Advantageous effects
The invention selects a specific protective matrix, and the thalli and the matrix are mixed before spray drying, so that the heat resistance and stress resistance activity of the bifidobacterium is improved, the survival rate of the bifidobacterium spray drying is obviously improved, and the survival rate of the bifidobacterium spray drying can reach more than 60 percent. Solves the problem of low survival rate of the bifidobacterium in the spray drying process.
The matrix before spray drying not only can play a role in improving the survival rate of the bifidobacteria, but also can ensure that the dried bifidobacteria powder has good fluidity, thereby facilitating the subsequent preparation processing.
The bifidobacterium powder prepared by the method is used as a raw material, a proper amount of milk powder, inulin, mannitol and silicon dioxide are added as auxiliary materials, and the powder packaged in a single dose is prepared by applying modern industrial technology.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to specific embodiments, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Bifidobacterium species: hansen BB-12.
Inoculating Bacillus bifidus BB-12 in TPY culture medium at 5%, culturing at 37 + -1 deg.C for 24 hr under anaerobic condition, and culturing at 4 deg.C for 6000r min-1Centrifuging for 15min, discarding supernatant, washing thallus with sterilized distilled water, and centrifuging again under the same conditions to obtain Bifidobacterium thallus.
The obtained cells and the dry protectant were mixed uniformly at a weight/volume ratio (1:100 g/mL). The formula of the spray drying matrix comprises: 10g of skimmed milk powder, 7g of maltose, 1.5g of calcium hydrophosphate, 0.4g of poloxamer and 81.1g of water.
And (3) carrying out spray drying on the mixed solution obtained in the previous step, wherein the spray drying conditions are as follows: the mixed liquid of the thalli is dried by a spray dryer at the speed of 5ml/min, the inlet temperature is 140 ℃, and the outlet temperature is 55 ℃. Spray drying to obtain Bifidobacterium powder. Survival rate of bifidobacterium: 60.1 percent.
The survival rate of bifidobacteria after spray drying was calculated according to the following formula: survival rate of bifidobacterium (viable count after drying) (cfu g)-1Dry matter)/viable count before drying (cfu g)-1Dry matter) × 100%.
Example 2
The spray-dried matrix formula is: 10g of skimmed milk powder, 7g of trehalose, 1.5g of calcium hydrophosphate, 0.4g of poloxamer and 81.1g of water. Other operating conditions were the same as in example 1. Survival rate of bifidobacterium: 52.8 percent.
Example 3
The spray-dried matrix formula is: 10g of skim milk powder, 7g of sucrose, 1.5g of calcium hydrophosphate, 0.4g of poloxamer and 81.1g of water. Other operating conditions were the same as in example 1. Survival rate of bifidobacterium: and (5) 55.3%.
Example 4
The spray-dried matrix formula is: 10g of skimmed milk powder, 7g of maltose, 1.5g of magnesium chloride, 0.4g of poloxamer and 81.1g of water. Other operating conditions were the same as in example 1. Survival rate of bifidobacterium: 48.7 percent.
Example 5
The spray-dried matrix formula is: 10g of skimmed milk powder, 7g of maltose, 1.5g of magnesium chloride, 0.4g of poloxamer and 81.1g of water. Other operating conditions were the same as in example 1. Survival rate of bifidobacterium: 51.4 percent.
Example 6
The spray-dried matrix formula is: 10g of skimmed milk powder, 7g of maltose, 1.5g of sodium dihydrogen phosphate, 0.4g of poloxamer and 81.1g of water. Other operating conditions were the same as in example 1. Survival rate of bifidobacterium: 57.6 percent.
Example 7
The spray-dried matrix formula is: 10g of skim milk powder, 5g of maltose, 1.5g of calcium hydrophosphate, 0.5g of poloxamer 188 and 83g of water. Other operating conditions were the same as in example 1. The angle of repose of the spray-dried bifidobacterium powder is measured by the following method: the funnel is fixed at a certain height above the coordinate paper (the paper is placed on a horizontal table), materials are added from the funnel until the top of the formed stacking cone is just contacted with the bottom of the funnel, the diameter of the cone is measured, and the ratio of the height of the bottom of the funnel to the radius of the cone is used as a tangent value to calculate the angle of repose. The angle of repose of the powder obtained in example 7 was 36.53 degrees.
Example 8
The spray-dried matrix formula is: 10g of skim milk powder, 5g of maltose, 1.5g of calcium hydrophosphate, 0.5g of Tween 80 and 83g of water. Other operating conditions were the same as in example 7. The angle of repose of the powder obtained in example 8 was 37.88 degrees.
Example 9
The spray-dried matrix formula is: 10g of skim milk powder, 5g of maltose, 1.5g of calcium hydrophosphate, 0.5g of SDS and 83g of water. Other operating conditions were the same as in example 7. The angle of repose of the powder obtained in example 8 was 38.47 degrees.
From examples 1 to 6, it can be seen that the dual protective agents of maltose and calcium hydrogen phosphate are selected, so that the heat resistance and stress resistance activity of the bifidobacteria is improved, the spray drying survival rate of the bifidobacteria is remarkably improved, and the spray drying survival rate can reach more than 60%. Solves the problem of low survival rate of the bifidobacterium in the spray drying process.
From examples 7 to 9, it can be seen that the protective agent of the present invention, when added with poloxamer as a surfactant, has the effect of improving the survival rate of bifidobacteria, and can also provide the dried bifidobacteria powder with good fluidity, thereby facilitating the subsequent preparation processing.
EXAMPLE 10 Probiotics powder production Process
1. Preparation of raw materials: the bifidobacterium powder, the milk powder, the inulin, the mannitol and the silicon dioxide which are prepared by the method are strictly placed in a clean raw material workshop for standby after being subjected to outsourcing in a buffer room according to the procedure of entering a clean area of 10 ten thousand levels.
2. Pretreatment of raw materials and auxiliary materials: respectively sieving Bifidobacterium powder, milk powder, inulin, mannitol and silicon dioxide with 60 mesh sieve.
3. Weighing: according to the total formula amount and the formula proportion of each batch of ingredients, the bifidobacterium powder, the milk powder, the inulin, the mannitol and the silicon dioxide are accurately weighed, and the check is performed by a person skilled in the art.
4. Mixing: firstly, the bifidobacterium powder and the mannitol are gradually and uniformly mixed according to the equal quantity to obtain mixed powder B, and the mixed powder B, the milk powder, the inulin and the silicon dioxide are added into a three-dimensional mixer to be mixed for 30 minutes.
5. Subpackaging: subpackaging with a particle packaging machine according to the requirement, wherein each bag is 1.5 g.
6. And (3) outer packaging: and (3) putting the subpackaged samples into a small box of 10 bags/box in an outer packaging room, labeling, boxing and packaging.
7. And (4) checking: and (5) inspecting the finished product according to the delivery inspection requirement of the product.
8. Warehousing: and (5) warehousing and storing the finished product after the finished product is qualified.
The production data of different batches of probiotic powder are as follows:
Figure RE-GDA0002265216740000071
Figure RE-GDA0002265216740000081
the names, models and manufacturers of the main production equipment are as follows:
name of production facility Model number Manufacturer of the product
Multi-directional motion mixer HD100 type Jiangnan pharmaceutical machinery Co Ltd of Zhejiang Ruian
Automatic powder packaging machine F40Ⅱ HONDON PACKAGING & FOOD MACHINERY Co.,Ltd.
The main raw material of the probiotic powder is the bifidobacterium powder prepared by the invention, and the powder is prepared by adding a proper amount of auxiliary materials, mixing and directly subpackaging. Before subpackaging, the flowability of the prepared probiotic powder is considered by taking the repose angle as an investigation index, and the measurement result of the repose angle of the mixed powder is as follows:
Figure RE-GDA0002265216740000082
as can be seen from the results of the angle of repose data in the table above: the average value of the repose angle alpha of the mixed powder measured for six times is 35.47 degrees, and the requirement of directly subpackaging the powder can be met when the alpha is generally considered to be less than or equal to 40 degrees. Therefore, the probiotic powder of the invention is feasible by directly subpackaging the mixed raw materials and auxiliary materials, and the obtained product has good fluidity.

Claims (5)

1. A preparation method of bifidobacterium powder is characterized by comprising the following steps:
a. culturing Bifidobacterium in culture medium, and collecting thallus;
b. uniformly mixing the thalli and a spray drying matrix to obtain a thalli mixed solution;
c. spray drying, and collecting Bacillus bifidus powder;
wherein the spray-dried matrix of step b comprises: 7-13% by weight of skim milk; 5-10% by weight of maltose; 1-2% by weight of calcium hydrogen phosphate; 0.3-0.5% by weight of poloxamer; the balance of water;
the poloxamer is selected from poloxamer 188;
the spray drying conditions of the step c are as follows: b, drying the thallus mixed solution obtained in the step b by a spray dryer at the speed of 4-6ml/min, wherein the inlet temperature is 130-;
wherein the culture medium in the step a is TPY culture medium; the bifidobacterium strain is Chr.Hansen BB-12;
the specific operation of the step a comprises the following steps: inoculating Bifidobacterium into TPY culture medium at 5%, culturing at 37 + -3 deg.C for 16-24 hr under anaerobic condition, and culturing at 4 deg.C for 5000--1Centrifuging for 10-20min, discarding supernatant, cleaning thallus with sterilized distilled water, and centrifuging under the same condition to obtain Bifidobacterium thallus.
2. The method for preparing bifidobacterium powder as claimed in claim 1, wherein the spray-dried matrix of step b comprises: 10% by weight of skim milk; 7-8% parts by weight of maltose; 1.4-1.6% by weight of calcium hydrogen phosphate; 0.35 to 0.45 percent of poloxamer; the balance being water.
3. A powder of bifidobacterium bacteria obtainable by a process according to any one of claims 1 to 2.
4. A method for preparing probiotic powder, comprising the steps of:
1) preparation of raw materials: placing bifidobacterium powder, milk powder, inulin, mannitol and silicon dioxide prepared by the method according to claim 1 into a clean area of 10 ten thousand grade according to a procedure, and immediately placing the bifidobacterium powder, the milk powder, the inulin, the mannitol and the silicon dioxide into a clean raw material workshop for standby after a buffer room is taken out of a package;
2) pretreatment of raw materials and auxiliary materials: respectively sieving bifidobacterium powder, milk powder, inulin, mannitol and silicon dioxide with a 60-mesh sieve for later use;
3) weighing: accurately weighing bifidobacterium powder, milk powder, inulin, mannitol and silicon dioxide according to the total formula amount and the formula proportion of each batch of ingredients;
4) mixing: firstly, sequentially and uniformly mixing bifidobacterium powder and mannitol in equal quantity to obtain mixed powder B, adding the mixed powder B, milk powder, inulin and silicon dioxide into a three-dimensional mixer, and mixing for 25-35 minutes;
5) subpackaging: subpackaging with a particle packaging machine according to requirements;
6) and (3) outer packaging: filling the subpackaged samples into small boxes of 10 bags/box in an outer packaging room, labeling, boxing and packaging;
7) and (4) checking: inspecting the finished product according to the delivery inspection requirement of the product;
8) warehousing: and (5) warehousing and storing the finished product after the finished product is qualified.
5. The method for preparing probiotic powder according to claim 4, characterized in that the mixing time of step 4) is 30 min.
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Denomination of invention: Preparation process of a bifidobacterium powder and its probiotics

Effective date of registration: 20231227

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Pledgee: Industrial and Commercial Bank of China Limited Chaozhou Branch

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