CN108823129A - A kind of cultural method of probiotics - Google Patents
A kind of cultural method of probiotics Download PDFInfo
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- CN108823129A CN108823129A CN201810722848.3A CN201810722848A CN108823129A CN 108823129 A CN108823129 A CN 108823129A CN 201810722848 A CN201810722848 A CN 201810722848A CN 108823129 A CN108823129 A CN 108823129A
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- probiotics
- bacterium powder
- solid beverage
- lactobacillus
- powder
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- 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
- A23V2200/00—Function of food ingredients
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/147—Helveticus
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- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/515—Animalis
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Abstract
The invention discloses a kind of cultural methods of probiotics.Porous material is added in the culture medium containing the somatomedin for promoting growth of probiotics breeding, and the volume ratio of culture medium and porous material is 1：0.001–1：1000, then be inoculated with probiotics in the medium and cultivated, realize probiotics High Density Cultivation；Probiotics obtained can be used for preparing the products such as medicine, feed or environmental protection treatment.The present invention, which obtains probiotics, has the performances such as good heat resistance, acid resistance and drug resistance, has broad application prospects in fields such as medicine, feed, environmental protection.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of cultural method of probiotics.
Background technique
Probiotics, English mono- word of probiotics are translated into " for life " from Greek " for live " derivation, are
Viable bacteria containing physiological activity after oral or other administration modes take in quantity appropriate, can be colonized in when by body
Host simultaneously improves host's microecological balance, to play beneficial effect.Recent studies indicate that the intracorporal bacterium of people is up to
Bacterium in more than 1000 kinds, especially gastrointestinal tract is the abundantest, accounts for whole body micro-ecological bacterial sum 80% or so, is human body cell
10 times, it is known as " the second genome " of the mankind.These bacteriums facilitate human consumption's food, generate vitamin to prevent food
The disease that middle bacterium is induced, while stimulating immune system.With going deep into for research, find enteric microorganism in many chronic diseases
In disease and symptom, such as inflammation, obesity, diabetes, also cancer also play key effect.The quantity and gene dosage of microorganism
It is surprising, the strong health for influencing the mankind.These are found to be probiotic products in food, health care product, medicine, cultivation, environment, health
The application in the fields such as articles brings huge motive force.Currently, probiotics is widely used in Yoghourt, sour milk beverage, prebiotic
In the products such as bacteria powder, animal feed, environmental improvement, relatively complete and mature probiotics industry is formd.
It is worth noting that, probiotics could generate in host when probiotics only reaches certain quantity levels
Corresponding effect.So needing to guarantee contained prebiotic bacterium number living by the food of functional component, health care product or drug of probiotics
Amount is maintained at certain level.This demand proposes corresponding requirement to the production and preparation of probiotics.In the training of probiotics
During supporting, realizes High Density Cultivation, not only can satisfy the functional requirement of subsequent probiotic products, probiotics can also be improved
Production efficiency in production link reduces volume of culture, shortens the production cycle, reduces production cost.Therefore, probiotics is highly dense
Degree culture is the key technical problem in current probiotics exploitation and production.
In liquid fermentation system, Chinese invention patent (publication number：CN 101230323A) alkalinity is added during the cultivation process
Suffered stress material acid or oxygen, make finally to produce when salt and deoxidier are to eliminate lactobacillus acidophilus or bifidobacterium bifidum fermentation
Number of viable improves 2-4 times in product.Chinese invention patent (publication number：CN 101418270A) in the training of Lactobacillus casei Zhang
During supporting, by the method for the addition neutralizers such as NaOH, KOH or ammonium hydroxide and logical nitrogen, 10 are obtained11CFU/mL or more viable count
Lactobacillus casei freeze-dried vaccine powder.Hayakawa K et al. (J Ferment Bioeng, 1990,70,404) uses membrane technology
It is coupled with fermentor, is constantly filtered to remove the stress ingredient such as lactic acid, realizes that highest cell density is up to 1011CFU/mL, this method
Mass propgation base need to be expended, and is only limitted to laboratory level, is difficult to be expanded to production application scale.
In semisolid or solid fermentation system, Chinese invention patent (publication number：CN 105132321A) using by wheat bran and
The culture medium of the material compositions such as dregs of beans, inoculation enterococcus faecium carry out solid state fermentation, and the viable bacteria content in dry products is up to 3.8
×1010CFU/g。
Similar also Chinese patent (publication number：CN 101555510B；CN 101555511B；CN 103421710B；
CN 102907563B；CN 103289937B；CN 1212391C) it is all made of dregs of beans, wheat bran, corncob, rice bran, peanut shell, shallow lake
The agricultural wastes such as powder, corn flour, turf or agricultural product solid medium as main component, microbe inoculation carry out highly dense
Degree ferments, and the viable count in product is essentially 1011CFU/g.This kind of product is mainly used for the metabolism such as animal feed or progress drug
Product production.
Probiotics is the active microorganism to be played a role by improving the balance of host intestine microbial flora, probiotics
Beneficial effect to health is that Russian scientists Mctchnikoff is proposed first, he thinks that enteron aisle lactic acid bacteria can be by anti-
Only the growth of spoilage organisms and play the role of extend the service life.Up to the present, it has been found that probiotics can improve enteron aisle
Bacterial community inhibits pathogen, eliminates carcinogen, improves immunity of organisms, reduces cholesterol.The health of probiotics and human body
There is close relationship, probiotics is added in milk can improve digestive function, promote enterogastric peristalsis, mitigate human body lactose not
It is resistant to disease, prevents the diarrhea and indigestion of infant, medically, probiotics can be used for abdomen caused by acute rotavirus
It rushes down, constipation, negative interaction caused by colon functional disturbance and radiation treatment can also be used to antitumor and anticancer.Currently, often
Probiotics strain has Bifidobacterium, Bacillus acidi lactici and some chain coccobacillus.Bifidobacterium, Bacillus acidi lactici etc. are beneficial in enteron aisle
Bacterium has the function of a variety of biological antagonists, such as Bacillus acidi lactici lactose fermenters in vivo, generates a large amount of lactic acid, acetic acid, can reduce intestines
Road local ph improves micro-ecological environment, Escherichia coli, detection of Salmonella, streptococcus etc. in antibacterial or kill enteron aisle.Meanwhile it is low
PH value is conducive to intestines peristalsis, maintains normal physiological function, prevents the field planting of pathogenic bacteria.The tissue acid that lactic acid bacteria generates is blue to leather
The effect of family name's negative bacterium becomes apparent from, and there are also the works for inhibiting pathogen duplication for lactic acid bacteria itself.Bifidobacterium can generate a kind of unknown
Broad-spectrum antiseptic substance, this substance can inhibit the activity of the pathogens such as salmonella, Shigella, comma bacillus.Probiotics
Product can play the role of preventing and treating disease while having additional nutrients.
Probiotics as a kind of new type functional adding ingredient, then health care product or field of medicaments play it is more and more important
Effect, however since thallus is very sensitive, it is easy to be affected by the ambient and lose activity.It is this that probiotics, which is stored at room temperature,
The big technical problem of industry one.Therefore, probiotic products gradually obtain public favor, and the too product such as probiotic yogurt is because of storage
Transportation cost is high, and the shelf-life is short etc. to limit its sales range, and solid kind probiotic products then store conveniently.
It is main former that solid beverage, which refers to sugared, cream and dairy products, egg or egg products, fruit juice or alimentary vegetable extracts etc.,
Material adds the solid articles that every 100 grams of moisture content of finished products made of suitable auxiliary material or food additives are not higher than 5 grams, is in powder
Shape, graininess or bulk etc..With the continuous improvement of people's living standards, consumption scaling trend has been more and more obvious, from market
From the point of view of, be to adapt to consumer to various needs such as instant solid beverage taste, nutrition and health care, start to follow natural nutrition,
The good direction of the thought of back to nature, solid beverage towards component nutrient laden, diversification of varieties, functional diversities develops.
Chinese patent 201710385573.4 discloses a kind of production method of probiotics solid beverage, and this method includes benefit
Raw bacterium culture, probiotics embedding, freeze-drying, allotment, secondary embedding, heated-air drying and packaging storage step.The inventive method
Using lactic acid bacteria powder and inulin as primary raw material, and add functional sugar and fruit vegetable powder is mixed and made into probiotics solid beverage.The hair
It is bright to use ultrasonic wave assisted fermentation technology, alternating temperature Freeze Drying Technique and secondary embedding techniques, it substantially increases lactic acid bacteria and is making
In the intracorporal survival rate of people, survival ability and bioactivity during work and after edible.
Chinese patent 20161038905306 disclose a kind of draft yeast probiotics solid beverage and preparation method thereof and
Using the draft yeast probiotics solid beverage according to the group of mass percent meter by being grouped as follows：Probiotic powder is lyophilized
20~40%, prebiotics 5~20%, water-soluble dietary fiber 0.2~20%, Chinese herbal medicine powder 5~20%, sweetener 0~8%.
The draft yeast probiotics solid beverage, which has, to be adjusted intestinal flora balance, improves the multiple efficacies such as immunity of organisms, can be made
For health care product, have the effect of tonifying spleen moistening lung, quench one's thirst promote the production of body fluid, regulating qi-flowing for harmonizing stomach, it is sub- strong that insomnia and dreamful sleep, fatiguability etc. can also be improved
Health symptom.
Chinese patent 201710466714.5 discloses a kind of probiotics solid beverage and preparation method thereof, including raw material,
Auxiliary material and probiotic powder, the raw material gram include by weight：1.2 grams and 0.6 gram of stachyose of oligofructose, the auxiliary material raw material is pressed
Weight in grams includes：0.1 gram of yeast powder and 0.1 gram of strawberry powder, the probiotic powder include：B. animais 2,000,000,000, newborn bifid
Bacillus 8,000,000,000 and Lactobacillus rhamnosus 8,000,000,000；This kind of probiotics solid beverage is able to maintain that the bacterium colony in enteron aisle balances, can either
Diarrhea is eliminated in help, and can prevent constipation, plays good dual regulation to enteron aisle.
Helicobacter pylori is a kind of gram-Negative bacillus, as long as being chronic active gastritis, peptic gastric ulcer
The cause of disease, and it is in close relations with stomach malignancy, and the World Health Organization and International Union Against Cancer are classified as I class carcinogenic substance, existing
The more probiotics solid beverage of technical report is mostly the bacterium colony balance that enteron aisle can be maintained to pinch, but is made to anti-helicobacter pylori
The research of probiotics solid beverage is less, it is therefore desirable to provide a kind of with the prebiotic of disinfecting helicobacter pylori effect
Bacterium solid beverage.
Summary of the invention
In order to solve the problems, such as background technique, the purpose of the present invention is to provide a kind of methods of probiotics.
The technical solution adopted by the present invention is as follows：
A kind of cultural method of probiotics：
Porous material is added in the culture medium containing the somatomedin for promoting growth of probiotics breeding, culture medium and more
The volume ratio of Porous materials is 1：0.001–1：1000, then be inoculated with probiotics in the medium and cultivated, realize that probiotics is high
Density culture.
The culture is Anaerobic culturel, and the probiotics being inoculated with is the microorganism of anaerobism or amphimicrobian.
Anaerobic culturel is to cultivate 12-48h at 30-42 DEG C.
After culture, the probiotics gathers the surface or inside or surface and inside of porous material, obtains
Total viable count is up to 10 in product after drying11-1015CFU/g or more.Viable count is influenced by factors such as drying means, freezing
Viable count is higher in dry product.
Preparation-obtained probiotics has the performances such as good heat resistance, acid resistance and drug resistance.
The culture medium is liquid, semisolid or solid-state, and cultivating system can be Liquid Culture, Semi-solid cell culture and consolidate
Any one of three kinds of training methods of body culture, the porous material are solid-state.
The culture medium is YPD (saccharomycete), Medium of Bifidobacterium (bifidobacterium adolescentis, animal bifidobacteria, cream
Bifidobacterium, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium longum), MRS culture medium (acidophilus cream bar
Bacterium, Lactobacillus casei, lactobacillus delbruockii subspecies bulgaricus, Rui Shi lactobacillus, Yue Shi lactobacillus, lactobacillus plantarum, Luo Yishi
The double second of lactobacillus, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus paracasei, Lactococcus lactis subsp. lactis, Lactococcus lactis
Acyl subspecies, lactobacillus gasseri, Pediococcus, enterococcus spp, Wei Si Bordetella, honeybee ball category, Lactobacillus crispatus), SIIM0012
Culture medium (lactobacillus fermenti, Lactobacillus rhamnosus), M17 culture medium (streptococcus thermophilus), lactic acid bacteria culturing medium (Lactococcus lactis
Subsp. cremoris), Leuconostoc mesenteroides culture medium (leuconostoc mesenteroides subsp mesenteroides), propionic acid bacteria culture medium (Propionibacterium
Belong to), wort agar (Debaryomyces, candida, Torulopsis), blood agar culture-medium (Bacteroides, shuttle bar
Pseudomonas), tomato juice culture medium (wine Coccus), PDA potato, glucose agar medium (Penicillium), SICC0070 butyric acid
Bacterium culture medium (fusobacterium), TSA tryptose soya agar (Aerococcus), broth bouillon (bacillus), LB culture
Base (Micrococcus) etc..
Contain the somatomedin for promoting growth of probiotics breeding in the culture medium.It is numerous containing promotion growth of probiotics
The somatomedin grown such as whey fermentation liquid, collagen fermentation liquid, potato fermentation liquid, marine algae extract, mushroom fermentation
Liquid or extract, soya-bean milk, tomato juice, carrot juice, corn juice, Cordyceps extract, Ganodenna Lucidum P.E, dendrobium candidum extract
Object, angelica extract, ginkgo biloba p.e, malt extract, ginseng class extract, Astragalus Root P.E, Fructus Monordicae extract, chrysanthemum
Powder, yeast dextran, xylo-oligosaccharide, vitamin, amino acid, dopamine, phytic acid, fulvic acid, humic acid, magnesium salts, zinc salt, manganese
Salt, calcium salt, molysite, selenium salt etc. it is one or more.
The mean pore size of porous material in the culture medium is 1 nanometer -500 microns, and volumetric porosity is
5%-99.9% (v/v).The volume ratio that 5%-99.9% (v/v) refers to that the hole in porous material accounts for entire material is 5%-
99.9%.
The porous material is gel rubber material, aerogel material, foamed material, fibrous material, poromerics, mesoporous material
Material or large pore material etc..
The gel rubber material, aerogel material, foamed material and fibrous material average pore size from nano-grade size to
Micron order size differs；The average pore size of the poromerics less than 2 nanometers, receive for 2-50 by the average pore size of mesoporous material
The average pore size of rice, large pore material is greater than 50 nanometers.
The porous material is natural polysaecharides and its derivative, native protein class and its derivative, synthesis macromolecule
Homopolymer or one of materials such as copolymer, inorganic material or a variety of.
Natural polysaecharides such as chitin, chitosan, cellulose, lignin, glucan, agarose, oligofructose, soybean is low
Glycan, konjaku glucomannan, oligoisomaltose, xylo-oligosaccharide, oligomeric maltose, starch, dextrin, inulin, alginic acid and its
Salt, dextran, hyaluronic acid, algal polysaccharides, algal polysaccharide, spirulina polysaccharide, lichenin, lentinan, ganoderma lucidum polysaccharide,
Tremella polysaccharides, Arabic gum, pectin, guar gum, xanthan gum carrageenan etc. are from plant, animal, seaweed and microorganism
More carbohydrates and their derivatives.
Native protein class such as soybean protein, mung bean protein, wheat gluten, zein, glutelin, fibroin, spider
Silk-fibroin, collagen, lactalbumin, gelatin, casein sodium, fibrin, keratin etc. are from plant, animal and micro-
The albumen and its derivative of biology.
Synthesize high molecular homopolymer and the copolymer such as poly- hydroxyl of polyaminoacid, polyethylene glycol, polylactic acid, polylactic acid-
Acetate multipolymer, polyvinylpyrrolidone, polycaprolactone, poly beta-hydroxybutyrate, poly- β hydroxypentanoic acid etc..
Inorganic material such as perlite, montmorillonite, carbon black, white carbon black, ceramics etc..
The porous material is the one or more of material described above, but is not limited to the above material.
Probiotics used is the microorganism to people and (or) animal health, such as Bifidobacterium, bifidobacterium adolescentis, is moved
Object Bifidobacterium, bifidobacterium lactis, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium longum, lactobacillus
Category, lactobacillus acidophilus, Lactobacillus casei, Lactobacillus crispatus, lactobacillus delbruockii subspecies bulgaricus (lactobacillus bulgaricus), moral
Family name's Lactobacillus sp, lactobacillus fermenti, lactobacillus gasseri, Lactobacillus helveticus, Yue Shi lactobacillus, Lactobacillus paracasei, plant cream
Bacillus, lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius, streptococcus, streptococcus thermophilus, lactococcus, lactic acid ball
Bacterium lactic acid subspecies, lactococcus lactis subsp, Lactococcus lactis biacetyl subspecies, leuconostoc mesenteroides subsp mesenteroides, propionic acid
Bacillus, saccharomyces, Dbaly yeast number, candida, Torulopsis, Bacteroides, Fusobacterium, wine Coccus,
Peptostreptococcus, Penicillium, hammer category, fusobacterium, Aerococcus, Pediococcus, bacillus, pluton category, intestines
Coccus, Micrococcus, Wei Si Bordetella etc. it is one or more, but be not limited to the above strain.
In the culture medium, the probiotics bacterial liquid measure being inoculated on porous material is 0.1%-50% (v/v).0.1%-50%
(v/v) volume for referring to probiotics bacterial liquid addition is the 0.1%-50% of entire culture volume after being added.
The concentration of bacterium solution in the probiotics inoculation bacterium solution is 108-109CFU/mL。
Cultural method culture according to the present invention, strain is in the medium under multi-component promotion, growth and breeding rate
It is substantially improved, after culture, drying can be prepared by probiotic products.Total viable count in product is up to 1011-1015CFU/g
More than, it can be used as probiotics leaven or probiotics preparation.This probiotics has good heat resistance, resistance to acid and alkali and drug resistance
The performances such as property.
The present invention can be summarized as：
The object of the present invention is to provide a kind of high-activity probiotics solid beverages, by weight include：Resistance paste
40-60 parts smart, 5-15 parts of inulin, 5-15 parts of oligofructose, 2-8 parts of Lactobacillus helveticus bacterium powder, Lactobacillus rhamnosus bacterium powder 2-8
Part, 2-8 parts of B. animais bacterium powder, newborn 2-8 parts of Bifidobacterium Bifidum bacterium powder, 2-8 parts of lactobacillus acidophilus bacterium powder and not tally double qi bars
2-8 parts of bacterium bacterium powder.The each component interaction of high-activity probiotics solid beverage provided by the invention, can promote stomach compacted
It is dynamic, promote the balance of stomach flora, increase the single effect of each component as a result, helicobacter pylori can be removed preferably,
Increase the content of beneficial bacterium.The invention belongs to food technology fields, and in particular to a kind of solid beverage and preparation method thereof, more
Body it is related to a kind of high-activity probiotics solid beverage and preparation method thereof.
In view of the deficiencies of the prior art, the present invention provides a kind of high-activity probiotics solid beverages and preparation method thereof.
On the one hand, the present invention provides a kind of high-activity probiotics solid beverages, by weight include：Resistance paste
40-60 parts smart, 5-15 parts of inulin, 5-15 parts of oligofructose, 2-8 parts of Lactobacillus helveticus bacterium powder, Lactobacillus rhamnosus bacterium powder 2-8
Part, 2-8 parts of B. animais bacterium powder, newborn 2-8 parts of Bifidobacterium Bifidum bacterium powder, 2-8 parts of lactobacillus acidophilus bacterium powder and not tally double qi bars
2-8 parts of bacterium bacterium powder.
Preferably, the mass ratio of the resistant dextrin, Lactobacillus rhamnosus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder is 8：
1：1；
Preferably, the mass ratio of the oligofructose, Switzerland's cream bar bacillus bacterium powder and B. animais bacterium powder is 3：
1：2；
Preferably, the inulin, newborn Bifidobacterium Bifidum bacterium powder, lactobacillus acidophilus bacterium powder mass ratio be 2：1：1.
In some embodiments for more having choosing, the high-activity probiotics solid beverage further includes kudzu-vine root powder and black
Bean powder, pueraria lobata include 12% flavone compound, such as Puerarin, daizeol glycosides, Peanut Squares nutritional ingredient, also contain egg
The minerals such as white matter, amino acid, sugar and the necessary iron of human body, calcium, copper, selenium, are rare tonics suitable for people of all ages, there is " people in thousand
The good reputation of ginseng ".Semen sojae atricolor also known as black beans, it is sweet in flavor mild-natured, there is high protein, characteristic low in calories, have it is anti-oxidant, anti-aging
Effect.
Therefore, as a kind of preferred implementation method, above-mentioned high-activity probiotics solid beverage, by weight
Further include：5 parts of kudzu-vine root powder, 8 parts of black bean powder.
In embodiment in the present invention, the Lactobacillus helveticus bacterium powder is Switzerland bacillus R52 bacterium powder；The sandlwood
Sugared lactobacillus bacterium powder is Lactobacillus rhamnosus R1l bacterium powder；The B. animais bacterium powder is B. animais BB-12 bacterium
Powder；The newborn Bifidobacterium Bifidum bacterium powder is newborn Bifidobacterium Bifidum Bi-07 bacterium powder；The lactobacillus acidophilus bacterium powder is lactobacillus acidophilus
NCFM bacterium powder；The not tally Bifidobacterium Bifidum bacterium powder is bifidobacterium bifidum Bb-06 bacterium powder.
On the other hand, it the present invention provides a kind of preparation method of high-activity probiotics solid beverage, specifically includes following
Step：
(1) the Lactobacillus helveticus bacterium powder of formulation dosage, Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder, cream is double
Qi bacillus bacterium powder, lactobacillus acidophilus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder after mixing evenly, obtain mixture A；
(2) inulin, resistant dextrin and mixture A are mixed, after mixing evenly, obtains mixture B；
(3) obtained mixture B is mixed with remaining raw material in formula, after mixing evenly, obtains mixture D；
(4) mixture D for obtaining step (3) is freeze-dried to get the probiotics solid beverage is arrived.
The revolving speed stirred in the step (1) is 5-15r/min, and the time of stirring is 30-50min.
The revolving speed stirred in the step (2) is 20-30r/min, and the time of stirring is 20-40min.
The revolving speed stirred in the step (3) is 35-45r/min, and the time of stirring is 10-30min.
The condition of freeze-drying is in the step (4)：Vacuum degree 0.095-0.01Mpa keeps 3-4h at 5-10 DEG C
Afterwards, after keeping 2-3h at 0-5 DEG C, after keeping 1-2h at -5-0 DEG C, after keeping 1-2h under -10-0 °, at -20--10 DEG C
After lower holding 1-2h, 1-2h is kept at -30--20 DEG C.
Lactobacillus helveticus bacterium powder, Lactobacillus rhamnosus bacterium powder, B. animais bacterium without special instruction, in the present invention
Viable count in powder, newborn Bifidobacterium Bifidum bacterium powder, lactobacillus acidophilus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder is 2x1011cfu/g。
Beneficial effects of the present invention are：
(1) high-activity probiotics solid beverage provided by the invention, including：Resistant dextrin, inulin, oligofructose, Switzerland
Lactobacillus bacterium powder, Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder, newborn Bifidobacterium Bifidum bacterium powder, lactobacillus acidophilus bacterium powder and
Not tally Bifidobacterium Bifidum bacterium powder, interacts each other, can promote enterogastric peristalsis, promotes the balance of stomach flora, increases
The result of the single effect of each component.
(2) high-activity probiotics solid beverage provided by the invention can preferably remove helicobacter pylori, promote intestines
The increase of organic acid and short chain fatty acids in road, reduces the pH value of stomach, and acidity is caused to reduce and inhibit helicobacter pylori, increases
The content of beneficial bacterium.
(3) high-activity probiotics solid beverage provided by the invention can mitigate inflammation, promote digestion, strengthen immunity.
Specific embodiment
Explanation："-" in all tables of the present invention indicates that this content is not present.
A kind of high-activity probiotics solid beverage, by weight include：40-60 parts of resistant dextrin, inulin 5-15
Part, 5-15 parts of oligofructose, 2-8 parts of Lactobacillus helveticus bacterium powder, 2-8 parts of Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder
2-8 parts of 2-8 parts, newborn 2-8 parts of Bifidobacterium Bifidum bacterium powder, 2-8 parts of lactobacillus acidophilus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder.
In embodiment in the present invention, Lactobacillus helveticus bacterium powder is Switzerland bacillus R52 bacterium powder；The rhamnose cream bar
Bacterium bacterium powder is Lactobacillus rhamnosus R1l bacterium powder；The B. animais bacterium powder is B. animais BB-12 bacterium powder；Institute
The newborn Bifidobacterium Bifidum bacterium powder stated is newborn Bifidobacterium Bifidum Bi-07 bacterium powder；The lactobacillus acidophilus bacterium powder is lactobacillus acidophilus NCFM
Bacterium powder；The not tally Bifidobacterium Bifidum bacterium powder is bifidobacterium bifidum Bb-06 bacterium powder.
In the present invention in some embodiments, the high probiotics solid beverage of above-mentioned high activity further includes kudzu-vine root powder and semen sojae atricolor
Powder.
The preparation method of the high-activity probiotics solid beverage is：
(1) the Lactobacillus helveticus bacterium powder of formulation dosage, Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder, cream is double
Qi bacillus bacterium powder, lactobacillus acidophilus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder after mixing evenly, obtain mixture A；
(2) inulin, resistant dextrin and mixture A are mixed, after mixing evenly, obtains mixture B；
(3) obtained mixture B is mixed with remaining raw material in formula, after mixing evenly, obtains mixture D；
(4) mixture for obtaining step (3) is freeze-dried to get the probiotics solid beverage is arrived.
The revolving speed stirred in step (1) is 5-15r/min, and the time of stirring is 30-50min.
The revolving speed stirred in step (2) is 20-30r/min, and the time of stirring is 20-40min.
The revolving speed stirred in step (3) is 35-45r/min, and the time of stirring is 10-30min.
The condition of freeze-drying is in step (4)：Vacuum degree 0.095-0.1Mpa, after keeping 3-4h at 5-10 DEG C,
After keeping 2-3h at 0-5 DEG C, after keeping 1-2h at -5-0 DEG C, after keeping 1-2h under -10-0 °, kept at -20--10 DEG C
After 1-2h, 1-2h is kept at -30--20 DEG C.
By adjusting the specific experiment parameter in basic embodiment, embodiment 1-10 as shown in Table 1 has been obtained.
A kind of high-activity probiotics solid beverage of comparative example 1 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that without Lactobacillus helveticus R52 bacterium powder, preparation in formula
Method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 2 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that without Lactobacillus rhamnosus R1l bacterium powder, system in formula
Preparation Method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 3 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that be free of B. animais BB-12 bacterium powder in formula,
Preparation method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 4 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that without newborn Bifidobacterium Bifidum Bi-07 bacterium powder, system in formula
Preparation Method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 5 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that without lactobacillus acidophilus NCFM bacterium powder, system in formula
Preparation Method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 6 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that be free of bifidobacterium bifidum Bb-06 bacterium powder in formula,
Preparation method is the same as embodiment 3.
A kind of high-activity probiotics solid beverage of comparative example 7 and preparation method thereof
Formula is substantially with embodiment 3, with embodiment 3 the difference is that progress primary freeze drying, freezing in step (4)
The dry time is -25 DEG C, and cooling time is 11 hours.
Experimental example 1
The high-activity probiotics solid beverage of embodiment 1-10 and comparative example 1-7 preparation is subjected to external inhibition pylorus spiral
Bacillus test, specific step is as follows：
1, material
Bacterial strain H.CagA (+) pylori is international standard bacterial strain NCTC11637, is CagA (+) and VacA (+).
2, method
In an aseptic environment, the 5g probiotics solid beverage for weighing embodiment 1-10 and comparative example 1-7, is dissolved in respectively
It is 2.0 with hydrochloric acid conditioning solution pH in 200ml sterile water, and 40 DEG C of constant temperature stir 2h, filtering, filtrate rotary evaporation to volume
Less than 50ml, 100ml then is settled to sterile water.It then is 1 by doubling dilution:40.It is broken into filter paper punch several straight
Diameter is the circular filter paper piece of 6mm, 120 DEG C of high pressure steam sterilizations.Under gnotobasis, the filter paper of sterilizing is clamped respectively with aseptic nipper
Piece, it is spare after moisture evaporation on every filter paper plus 10 μ l composition solutions.
(2) jejunum campylobacter agar medium 450g is weighed, distilled water 1000ml, heating stirring dissolution, high temperature and pressure is added
Sterilizing, is cooled to 50-60 DEG C.100ml is added and takes off fiber sheep blood, stirs evenly, topples over sterile petri dish.By H.pylori in three
3d is cultivated in gas incubator, is eluted with brucella broth and is configured to suspension.H. pylori bacteria concentration is detected by microplate reader,
It is prepared as 108The brucella broth bacterium solution of cfu/ml, is applied on plate.
(3) film-making diffusion method is used, vernier caliper measurement antibacterial circle diameter is passed through.
2, experimental result is as shown in table 2
From Table 2, it can be seen that the probiotic composition provided in 1-10 of the embodiment of the present invention has helicobacter pylori
Good fungistatic effect, much higher than the probiotic composition provided in comparative example 1-10.Wherein by adjusting resistance in embodiment 4
The mass ratio of dextrin, Lactobacillus rhamnosus R1l bacterium powder and bifidobacterium bifidum Bb-06 bacterium powder is 8:1:1, probiotics can be improved
Bacteriostatic activity of the composition to helicobacter pylori.
Experimental example 2
1, material:Male mice is weighed, every group 10, the high-activity probiotics provided according to 1-10 of the embodiment of the present invention
Solid beverage weighs to mouse, practices blood stomach-filling 10 days to mouse according to 1.67g/kg.bw respectively, blank group sterile diluent
Tank stomach.Before giving tested material, take stool in mice 0.1g, add 10 times of sterile saline dilutions, calculate every gram it is wet just in
Bacterium number takes the laggard line number of logarithm according to statistics, sterile to take excrement after giving tested material 24 for the last time, detects intestinal flora,
Method is same as above, as a result and shown in table 3：
From table 3 it is observed that the high-activity probiotics solid beverage that 1-10 of the embodiment of the present invention is provided, through mouse gavaging
Afterwards, the content of Bifidobacterium Bifidum and Bacillus acidi lactici is higher than gavage before, and the high-activity probiotics solid beverage provided in comparative example 1-7
In Bifidobacterium Bifidum and the quantity of Bacillus acidi lactici do not obviously increase, or even reduce, illustrate that the probiotics that this formula provides is consolidated
Body beverage has the function of improving intestinal colony.Wherein embodiment 5 and embodiment 3 the difference is that, have adjusted oligofructose, auspicious
The mass ratio of scholar's lactobacillus R52 bacterium powder and B. animais BB-12 bacterium powder is 3:1:2, mouse intestinal can be significantly increased
The content of middle Bacillus acidi lactici and Bifidobacterium has statistical difference (P<0.05).The wherein difference of embodiment 6 and embodiment 3
The mass ratio for being to adjust inulin, newborn Bifidobacterium Bifidum Bi-07 bacterium powder and lactobacillus acidophilus NCFM bacterium powder is 2:1:1, embodiment 6 mentions
The enterococcal content of mouse intestinal can be significantly reduced in the high-activity probiotics solid beverage of confession, has statistical difference (P<
0.05).For embodiment 10 with embodiment 7 the difference is that containing black bean powder and kudzu-vine root powder, the high activity that embodiment 10 provides is prebiotic
The content of the C.perfringens of mouse intestinal can be significantly reduced in bacterium solid beverage, has statistical difference (P<0.05).
Only as described above, is only a preferred embodiment of the present invention, when the model that cannot limit implementation of the invention with this
It encloses, i.e., generally according to simple equivalent changes and modifications made by content documented by the claims in the present invention and description of the invention,
All it is still covered by the claims of the invention.It is searched in addition, abstract part and title are intended merely to auxiliary patent document
It seeks and being used, the interest field being not intended to limit the invention.
Claims (10)
1. a kind of cultural method of probiotics, it is characterised in that：Containing the somatomedin for promoting growth of probiotics breeding
Add porous material in culture medium, the volume ratio of culture medium and porous material is 1：0.001–1：1000, then connect in the medium
Kind probiotics is simultaneously cultivated, and realizes probiotics High Density Cultivation；The culture is Anaerobic culturel, and the probiotics being inoculated with is to detest
The microorganism of oxygen or amphimicrobian；After culture, the probiotics gathers surface or inside or the table of porous material
Face and inside, total viable count is up to 5 × 10 in the product after being dried11-3×1015CFU/g or more；The culture medium is
Liquid, semisolid or solid-state, the porous material are solid-state.
2. the cultural method of probiotics as described in claim 1, it is characterised in that：In the culture medium, connect on porous material
The probiotics bacterial liquid measure of kind is 0.1%-50%；The pore size of the porous material is 1 nanometer -500 microns, volume hole
Rate is 5%-99.9%；It is characterized in that：The porous material is gel rubber material, aerogel material, foamed material, undulation degree
One of materials such as material, poromerics, mesoporous material or large pore material are a variety of；The porous material is natural polysaccharide
The materials such as class and its derivative, native protein class and its derivative, the high molecular homopolymer of synthesis or copolymer, inorganic material
One of or it is a variety of.
3. a kind of high-activity probiotics solid beverage, by weight include：40-60 parts of resistant dextrin, 5-15 parts of inulin,
5-15 parts of oligofructose, 2-8 parts of Lactobacillus helveticus bacterium powder, 2-8 parts of Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder 2-8
2-8 parts of part, newborn 2-8 parts of Bifidobacterium Bifidum bacterium powder, 2-8 parts of lactobacillus acidophilus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder.
4. high-activity probiotics solid beverage as claimed in claim 3, it is characterised in that：The resistant dextrin, rhamnose
The mass ratio of lactobacillus bacterium powder and not tally Bifidobacterium Bifidum bacterium powder is 8：1：1.
5. high-activity probiotics solid beverage as claimed in claim 3, it is characterised in that：The oligofructose, Switzerland's cream
The mass ratio of bar bacillus bacterium powder and B. animais bacterium powder is 3：1：2.
6. high-activity probiotics solid beverage as claimed in claim 3, it is characterised in that：The inulin, newborn Bifidobacterium Bifidum
The mass ratio of bacterium powder and lactobacillus acidophilus bacterium powder is 2：1：1.
7. high-activity probiotics solid beverage as claimed in claim 3, it is characterised in that：The high-activity probiotics solid
Beverage further includes kudzu-vine root powder and black bean powder.
8. the preparation method of the high-activity probiotics solid beverage as described in claim 3-7 any one, it is characterised in that：Tool
Body includes the following steps：
(1) by the Lactobacillus helveticus bacterium powder of formulation dosage, Lactobacillus rhamnosus bacterium powder, B. animais bacterium powder, the double qi bars of cream
Bacterium bacterium powder, lactobacillus acidophilus bacterium powder, not tally Bifidobacterium Bifidum bacterium powder after mixing evenly, obtain mixture A；
(2) by inulin, resistant dextrin and mixture A mixing, after mixing evenly, mixture B is obtained；
(3) obtained mixture B is mixed with remaining raw material in formula, after mixing evenly, obtains mixture D；
(4) mixture for obtaining step (3) is freeze-dried to get the probiotics solid beverage is arrived.
9. the preparation method of high-activity probiotics solid beverage as claimed in claim 8, it is characterised in that：The step (1)
The revolving speed of middle stirring is 5-15r/min, and the time of stirring is 30-50min；The revolving speed stirred in the step (2) is 20-30r/
Min, the time of stirring are 20-40min；The revolving speed stirred in the step (3) is 35-45r/min, and the time of stirring is 10-
30min。
10. the preparation method of high-activity probiotics solid beverage as claimed in claim 8, it is characterised in that：The step (4)
The condition of middle freeze-drying is：Vacuum degree 0.095-0.01Mpa keeps 2-3h at 0-5 DEG C after keeping 3-4h at 5-10 DEG C
Afterwards, after keeping 1-2h at -5-0 DEG C, after keeping 1-2h under -10-0 °, after keeping 1-2h at -20--10 DEG C, in -30--
1-2h is kept at 20 DEG C.
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| CN201810722848.3A CN108823129B (en) | 2018-07-04 | 2018-07-04 | A kind of high-activity probiotics solid beverage with disinfecting helicobacter pylori effect |
| PCT/CN2019/094325 WO2020007276A1 (en) | 2018-07-04 | 2019-07-02 | Highly active probiotic solid beverage having resistance to helicobacter pylori |
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| CN111616347A (en) * | 2020-06-12 | 2020-09-04 | 四川省食品发酵工业研究设计院 | Method and container for making self-fermented Sichuan pickle-flavored sauce |
| CN112353907A (en) * | 2020-11-23 | 2021-02-12 | 苏州微克生活科技有限公司 | Probiotic preparation with helicobacter pylori antagonism function and application thereof |
| WO2022095287A1 (en) * | 2020-11-06 | 2022-05-12 | 拉格比(广东)健康科技有限公司 | Probiotic drops for improving allergic constitution |
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