CN108523137A - A kind of preparation method of the probiotic granulate of acid and bile salt tolerance - Google Patents

A kind of preparation method of the probiotic granulate of acid and bile salt tolerance Download PDF

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Publication number
CN108523137A
CN108523137A CN201810354767.2A CN201810354767A CN108523137A CN 108523137 A CN108523137 A CN 108523137A CN 201810354767 A CN201810354767 A CN 201810354767A CN 108523137 A CN108523137 A CN 108523137A
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acid
preparation
lactobacillus
mixed
probiotic
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陶纯长
胡小丽
朱建国
姜甜
方曙光
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Jiangsu Wecare Biotechnology Co Ltd
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Jiangsu Wecare Biotechnology Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/015—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/256—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin from seaweeds, e.g. alginates, agar or carrageenan
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/20—Agglomerating; Granulating; Tabletting
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/123—Bulgaricus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/143—Fermentum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/145—Gasseri
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/147—Helveticus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/151—Johnsonii
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/165—Paracasei
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/173—Reuteri
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/175—Rhamnosus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/181—Salivarius
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/513—Adolescentes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/517—Bifidum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/519—Breve
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum

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Abstract

The present invention relates to field of probiotic bacteria more particularly to a kind of preparation methods of the probiotic granulate of acid and bile salt tolerance.Include the following steps：S1, probiotics bacterium mud is uniformly mixed with protective agent, is slowly stirred to form emulsion；S2, emulsion and colloidal solution are added to the colloidal solution configured after mixing, stirring makes it be uniformly mixed, is slowly stirred to form mixed liquor；S3, it mixed liquor is slowly injected into calcium chloride solution carries out cross-linking reaction, high shear agitation is carried out after the injection of whole mixed liquors, be then allowed to stand to obtain Mixed Microbes slurry；S4, Mixed Microbes slurry is obtained into the solid that water content is less than 5% by freeze-drying, is crushed whole grain using crushing and pelletizing machine and obtains the probiotic granulate of 20 50 mesh；Protective agent includes trehalose, glycerine and converted starch, and colloidal solution includes sodium alginate；The probiotic granulate that the preparation method of the present invention obtains reaches the holding activity that enteron aisle can be largely to sour and cholate with good tolerability, by stomach.

Description

A kind of preparation method of the probiotic granulate of acid and bile salt tolerance
Technical field
The present invention relates to field of probiotic bacteria more particularly to a kind of preparation methods of the probiotic granulate of acid and bile salt tolerance.
Background technology
Probiotics is a kind of bacterium that facilitation can be played to health, common probiotics include Bacillus acidi lactici, Bifidobacterium and streptococcus etc..For example, Bacillus acidi lactici is mainly colonized in human small intestine, it is big that Bifidobacterium is mainly colonized in human body If enteral can play the role of comprehensive adjustment using the strain of compounding to human body.The organic acid of these probiotics metabolism generation, Polysaccharide, bacteriocin and special enzyme system etc. have to human body adjust stomach colony balance, improve immunity, anti-aging, improve constipation, Diarrhea and allergic symptom and other effects.
Modern humans' living standard rises, and the pollution of environment, abuse of drug etc. are to the microecological balance of human body and immune Power causes many negative effects, and the crowd in inferior health is also increasing.And targetedly the edible food containing probiotics can To adjust the balance of human internal environment, autoimmunity is improved.
Currently, probiotics preparation is mostly deposited in the form of pulvis, tablet, hard capsule, granule etc. in food and field of health care products By the control of packaging material and environmental condition, further requirement could reached in product viable count stability, and in mouth It is easily interfered by stress such as alimentary canal, gastric juice, cholate during clothes, is easy to lose activity before entering small intestine, losing it should have Probiotic effects, cause it is practical fail to reach requirement in host intestine release action amount, so needing in product forms and preparation Process orientation is studied, and discussion can resist host's gastric juice, cholate stress, carry out the prebiotic becteriums product shape of enteron aisle controlled release Formula.
Invention content
There is good tolerability to acid and cholate, leads in order to solve the above technical problems, the object of the present invention is to provide one kind Cross the benefit that stomach reaches active, beneficial function to ensure probiotics the acid and bile salt tolerance of the holding that enteron aisle can be largely The preparation method of raw bacterium particle.
The present invention provides a kind of preparation method of the probiotic granulate of aforementioned acid and bile salt tolerance, includes the following steps：
S1, by probiotics bacterium mud and protective agent with 1:4 to 4:1 ratio is uniformly mixed, and is slowly stirred 10-40min and is formed Emulsion；
S2, by emulsion and colloidal solution with 1:2 to 8:The colloidal solution configured is added in 1 ratio, and stirring keeps its mixing equal It is even, it is slowly stirred 5-10min, forms mixed liquor；
S3, the calcium chloride solution that 0.25 to 1 times of the mass concentration that mixed liquor is slowly injected into its quality is 0.2%-5% Middle carry out cross-linking reaction carries out high shear agitation 2-10min after the injection of whole mixed liquors, is then allowed to stand 5-20min, obtains Mixed Microbes are starched；
S4, by Mixed Microbes slurry by freeze-drying obtain water content be less than 5% solid, using crushing and pelletizing machine by its into Row crushing and pelletizing obtains the probiotic granulate of 20-50 mesh；
Wherein, the protective agent includes trehalose, glycerine and converted starch, and the colloidal solution includes sodium alginate.
It should be noted that the sodium alginate in colloidal solution, which crosslinks to react with calcium chloride, can form macromolecular polymeric Cohesion occurs in low ph conditions (such as gastric juice) for object, the polymer, protects probiotic granulate non-inactivation, and in higher pH rings It is disintegrated in border (such as intestinal juice), discharges probiotic granulate therein.Protective agent then can effectively protect probiotics in higher pH rings It is not inactivated by cholate in border (such as intestinal juice), finally to promote probiotics in intestinal colonisation.
It should be noted that in step S1, probiotics bacterium mud and protectant mixed proportion, protectant ingredient and The time is mixed, it will influence bacterium amount yield, the acid and bile salt tolerance of the obtained probiotic granulate of preparation method of the present invention Performance, specific detailed in Example.
It should be noted that in step S2, the mixed proportion of emulsion and colloidal solution, it will influence probiotic granulate and exist Embedding effect in the macromolecule polyalcohol that sodium alginate and calcium chloride are cross-linked to form will cause when colloid liquid hold-up is too small It is bad to embed effect, when colloid liquid hold-up is excessive, bacterium amount yield will be caused relatively low and the waste of embedded material.
It should be noted that step S3 in, the mixed proportion and calcium chloride solution of mixed liquor and calcium chloride solution it is dense Degree, it will polymer physicochemical property, reaction speed and the intensity for influencing embedded material after cross-linking reaction, when calcium chloride solution ratio When example is higher, reaction speed will be caused too fast, when calcium chloride solution concentration is higher, cross-linking reaction excessive velocities, institute will be caused It is inhomogenous to obtain Mixed Microbes slurry reaction result.
The patent of invention that Chinese Patent Application No. is 200610010518.9 discloses a kind of beneficial bacteria of intestinal tract embedding protection Method, however by experimental results demonstrate the microbial bacterial agent Pa after freeze-drying can be redissolved after being mixed into emulsified solution B1, become Uniform mixture can't form double-layer embedment structure, after being blended into sodium alginate soln, can remix uniformly mixed at one Close liquid；In addition, the live probiotic bacteria preparation finally obtained in a manner of natural drying, it is also difficult to which industry is realized in mass application Change.
The patent of invention that Chinese Patent Application No. is 201610881048.7 discloses a kind of probiotic microcapsule and its system Preparation Method, however, using vegetable oil emulsifying manner, industrialized production also brings inconvenience, is actually also difficult to industrialization.
Further, it includes trehalose 20-50%, glycerine 2-10%, sucrose 2- that agent solution is protected described in step S1 10%, converted starch 2-10%, vitamin 0.1-2%, amino acid 1-8%, skimmed milk powder 2-20%.
Specifically, the colloidal solution includes 5% sodium alginate.
Specifically, the vitamin includes one or more in vitamin B, vitamin C, vitamin D, vitamin E.
Specifically, the skimmed milk powder includes ox skimmed milk powder, sheep skimmed milk powder, one kind in horse skimmed milk powder or more Kind.
Specifically, the amino acid includes methionine, tyrosine, arginine, cysteine, valine, histidine, bright ammonia It is one or more in acid, glycine, alanine, glutamic acid, lysine.
Specifically, the probiotics described in step S1 includes lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, sandlwood Sugared lactobacillus, lactobacillus reuteri, Lactobacillus paracasei, lactobacillus fermenti, lactobacillus gasseri, Yue Shi lactobacillus, Switzerland's breast bar Bacterium, Lactobacillus salivarius, lactobacillus bulgaricus, bifidobacterium longum, bifidobacterium breve, bifidobacterium lactis, bifidobacterium adolescentis, two It is one or more in discrimination Bifidobacterium, streptococcus thermophilus.
According to the above aspect of the present invention, the present invention has at least the following advantages：Feed liquid is prepared with sodium alginate mixed active ingredient, so Gel in calcium chloride solution is instilled afterwards detaches drying after particle, here it is the micro-embedded methods of common enteric at particle.Due to logical It is larger (being typically larger than 2.5mm) to cross the particle that dripping feed liquid granulating obtains, so developing emulsion process again, for example feed liquid is emulsified Into vegetable oil, then add calcium chloride solution, the particle of very little will be obtained, as grain size is less than 0.05mm.
Enteric coated particles probiotics uses in industrialization generally as additive, therefore higher to Particle size requirements, such as Otherwise 0.2-0.5mm cannot be mixed in compounding to end product.So the bulky grain of dropping preparation method production can not just use；So And emulsion process, as a result of vegetable oil, industrialized production also brings inconvenience, is actually also difficult to industrialization.Therefore, such as What obtains the smaller probiotic granulate of grain size by relatively simple production technology, becomes the developing direction of probiotics industry.
In the present invention, feed liquid and sodium alginate soln are mixed, are then added in calcium chloride solution, at this time in mixed liquor The gel piece that will produce bulk can form uniform little particle by uniform to the mixture high speed shear containing gel piece, freeze 20-50 mesh probiotic granulates can be obtained in crushing and pelletizing again after dry.Since the material after high speed shear has been formed substantially in fact Even little particle gel, although may have in a liquid it is viscous, freeze-drying after again crushing and pelletizing, be easy to get 20-50 purposes Grain；Crushing and pelletizing is difficult to obtain the little particle of 20-50 mesh after common feed liquid freeze-drying.
In addition, the present invention provides a kind of optimal production technology, reasonably protect agent prescription, work that can be relatively simple Skill obtains the probiotic granulate of preferable acid and bile salt tolerance, it can be achieved that industrialization.
Above description is only the general introduction of technical solution of the present invention, in order to better understand the technical means of the present invention, And can be implemented in accordance with the contents of the specification, it is described in detail as after with presently preferred embodiments of the present invention below.
Specific implementation mode
With reference to embodiment, the embodiment of the present invention is furthur described in detail.Following embodiment is used for Illustrate the present invention, but is not limited to the scope of the present invention.
Embodiment one
The present embodiment provides a kind of preparation method of the probiotic granulate of acid and bile salt tolerance by taking lactobacillus acidophilus as an example, and It detects that different protective agents are acidproof for probiotic granulate, influence of bile tolerance performance, includes the following steps：
S1, configuration protection agent solution, sterilizing postcooling is for use to room temperature, and substance used meets Food Inspection standard； The sodium alginate colloidal solution of configuration quality a concentration of 5%, sterilizing postcooling are for use to room temperature；
S2, by lactobacillus acidophilus bacterium mud with protection agent solution with 1：1 ratio is uniformly mixed, and is slowly stirred 30min, is formed Emulsion；
S3, with 2:Emulsion, is added the colloidal solution configured by 1 ratio, and stirring makes it be uniformly mixed, is slowly stirred 5min forms bacterium glue mixed liquor；
S4, bacterium glue mixed liquor is slowly injected into its quality 0.25 times of mass concentration be 2% calcium chloride solution in into Row cross-linking reaction carries out high shear agitation 5min after the injection of whole mixed liquors, is then allowed to stand 10min, obtains Mixed Microbes slurry；
S5, by Mixed Microbes slurry by freeze-drying obtain water content be less than 5% solid, using crushing and pelletizing machine by its into Row crushing and pelletizing obtains the probiotic granulate of 20-50 mesh.
It is specific as shown in table 1 the present embodiment provides three kinds of experiment group of formula for protective agent：
The different protectant formulas of table 1
Correspondingly, using formula 1, being formulated 2 and being formulated 3 and be pulled through the probiotic granulate that above-mentioned test method obtains, respectively For particle 1, particle 2 and particle 3.
The present embodiment also provides a kind of probiotic granulate of control group, and preparation method includes the following steps：
S1, configuration protective agent solution sterilization are for use, and protective agent solution formula is：It is 20% trehalose, 2% sucrose, 2% sweet Oil, 2% converted starch.
S2, by lactobacillus acidophilus bacterium mud and the protection agent solution of configuration with 1：1 ratio is uniformly mixed and obtains emulsion, will Emulsion obtains the solid that water content is less than 5% by freeze-drying, and solid fungus block is crushed to obtain pair by pulverizer According to a group Freeze-dry Powder of Probioctics.It should be noted that control group freeze-dried powder, due to being lyophilized again without after high speed shear, grain size is difficult To reach the level of 20-50 mesh, there is a situation where that a large amount of particles are more than 50 mesh.
By aforementioned particles 1, particle 2, particle 3 and control group freeze-dried powder, acidproof and bile tolerance performance evaluation is carried out, specifically such as Under：
1) acid resistance is studied
Simulated gastric fluid (dilute hydrochloric acid solution of pH=3) is prepared, four kinds of probiotic granulates or freeze-dried powder are placed in, constant temperature is put into Shaking table, 180r/min vibrates 1h under the conditions of 37 DEG C, and per 20min, sampling weighs dissolving Mass Calculation rate dissolution rate, and calculates bacterium Body survival rate.
About particle or freeze-dried powder dissolution rate, the results are shown in Table 2, it is seen that particle 1, particle 2 and particle 3 have good Acid-fast ability does not almost dissolve, which can be effectively protected thalline, is obviously improved to acid-resisting ability.
Dissolution rate comparing result table of the different probiotic granulates of table 2 in simulated gastric fluid
About thalline survival rate, the results are shown in Table 3, it is seen that particle 1, particle 2 and particle 3 even across simulated gastric fluid at Reason, can also be effectively ensured the survival rate of wherein probiotics.
Thalline survival rate comparing result table after the different probiotic granulates of table 3 are handled in simulated gastric fluid
2) enteric performance study
Simulated intestinal fluid is prepared, 4 kinds of probiotic granulates are placed in intestinal juice, are put into constant-temperature table, 180r/min under the conditions of 37 DEG C 1h is vibrated, sampling calculates thalline survival rate per 20min, and the results are shown in Table 4, it is seen that particle 1, particle 2 and particle 3 are by artificial Intestinal juice processing, can be effectively ensured the survival rate of wherein probiotics.
Thalline survival rate comparing result table after the different probiotic granulates of table 4 are handled in simulated intestinal fluid
The above is only a preferred embodiment of the present invention, it is not intended to restrict the invention, it is noted that for this skill For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is several improvement and Modification, these improvements and modifications also should be regarded as protection scope of the present invention.

Claims (7)

1. a kind of preparation method of the probiotic granulate of acid and bile salt tolerance, it is characterised in that：The present invention provides a kind of aforementioned acidproof The preparation method of the probiotic granulate of bile tolerance, includes the following steps：
S1, by probiotics bacterium mud and protective agent with 1:4 to 4:1 ratio is uniformly mixed, and is slowly stirred 10-40min and is formed emulsification Liquid；
S2, by emulsion and colloidal solution with 1:2 to 8:The colloidal solution configured is added in 1 ratio, and stirring makes it be uniformly mixed, and delays Slow stirring 5-10min, forms mixed liquor；
S3, mixed liquor is slowly injected into its quality 0.25 to 1 times of mass concentration be 0.2%-5% calcium chloride solution in into Row cross-linking reaction carries out high shear agitation 2-10min after the injection of whole mixed liquors, is then allowed to stand 5-20min, is mixed Bacterium is starched；
S4, Mixed Microbes slurry is obtained into the solid that water content is less than 5% by freeze-drying, powder is carried out using crushing and pelletizing machine Broken whole grain obtains the probiotic granulate of 20-50 mesh；
Wherein, the protective agent includes trehalose, glycerine and converted starch, and the colloidal solution includes sodium alginate.
2. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：In step S1 The protection agent solution includes trehalose 20-50%, glycerine 2-10%, sucrose 2-10%, converted starch 2-10%, vitamin 0.1-2%, amino acid 1-8%, skimmed milk powder 2-20%.
3. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：The colloid Liquid includes 5% sodium alginate.
4. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：The dimension life Element includes one or more in vitamin B, vitamin C, vitamin D, vitamin E.
5. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：The degreasing Milk powder includes one or more in ox skimmed milk powder, sheep skimmed milk powder, horse skimmed milk powder.
6. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：The amino Acid includes methionine, tyrosine, arginine, cysteine, valine, histidine, leucine, glycine, alanine, paddy ammonia It is one or more in acid, lysine.
7. the preparation method of the probiotic granulate of acid and bile salt tolerance according to claim 1, it is characterised in that：In step S1 The probiotics includes lactobacillus plantarum, Lactobacillus casei, lactobacillus acidophilus, Lactobacillus rhamnosus, lactobacillus reuteri, pair Lactobacillus casei, lactobacillus fermenti, lactobacillus gasseri, Yue Shi lactobacillus, Lactobacillus helveticus, Lactobacillus salivarius, bulgarian milk In bacillus, bifidobacterium longum, bifidobacterium breve, bifidobacterium lactis, bifidobacterium adolescentis, bifidobacterium bifidum, streptococcus thermophilus It is one or more.
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Application publication date: 20180914