CN110521785A - Probiotics fermention functional food and its preparation - Google Patents
Probiotics fermention functional food and its preparation Download PDFInfo
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- CN110521785A CN110521785A CN201910931518.XA CN201910931518A CN110521785A CN 110521785 A CN110521785 A CN 110521785A CN 201910931518 A CN201910931518 A CN 201910931518A CN 110521785 A CN110521785 A CN 110521785A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C20/00—Cheese substitutes
- A23C20/02—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates
- A23C20/025—Cheese substitutes containing neither milk components, nor caseinate, nor lactose, as sources of fats, proteins or carbohydrates mainly containing proteins from pulses or oilseeds
-
- 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
- A23V2400/41—Pediococcus
- A23V2400/413—Acidilactici
-
- 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
- A23V2400/515—Animalis
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to microbial fermentation application field, it is related to a kind of probiotics fermention food and its preparation.It is using probiotics to soybean, almond, fructus cannabis, brush-cherry seed carries out what mixed bacterium stepwise fermentation obtained, with the health-care efficacies such as dry, ease constipation, defaecation are dropped, the probiotics is bacillus subtilis, animal bifidobacteria, bifidobacterium adolescentis, bifidobacterium longum, lactobacillus plantarum, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, Pediococcus acidilactici.
Description
Technical field:
The invention belongs to microbial fermentation application fields, relate to the use of probiotics fermention soybean, almond, fructus cannabis, Yu Li
The functional food of the productions such as benevolence and the preparation method and application of functional food.
Background technique:
In recent decades, science science and technology in China's is quickly grown, and the living standard of social style and features and the people all greatly improve, people
Have to balanced comprehensive intake nutrition and more and more clearly recognize, the demand to food already far beyond meet existence and
The concept of the scope of taste bud, health care is rooted in the hearts of the people, and becomes a kind of trend and habit gradually.Bean product is as a kind of
Time-honored high protein diet, vegetable protein contained therein are more suitable for compared to the animal protein in meat milk
In the enteron aisle of Chinese, conducive to absorbing, mild nourishing the stomach and nutrition is comprehensive.Bacillus acidi lactici and Bifidobacterium in healthy human body enteron aisle
Quantity accounting is all higher.Probiotics can stimulate intestines peristalsis, extend excrement in enteron aisle by the time, to be conducive to remove
Toxin in gastrointestinal tract maintains the normal function of enteron aisle.Stable probiotic dietary can alleviate intractable constipation within several weeks.
Series of studies shows that probiotics is treating a plurality of types of diarrhea, if adult is because of diarrhea, infections in infants abdomen caused by antibiotic
Rushing down etc. has good curative effect.This may be to enhance since after intake probiotics, stimulation intestinal tract immune system produces antibody
The barrier function of enteron aisle.
The nutritive value of soybean is very high, referred to as " king in beans ", is most pushed away by nutritionist in hundreds of natural foods
High food.Tempeh not only contains original rich nutrient in soybean, but also is acted on by microbial fermentation, again
Generating many kinds has in the production of functional object tempeh of health-care effect human body, because participating in main micro- life of fermentation
The type of object is different, and product respectively has special favor.Wherein bacterium is present in the environment such as air, soil, water, extensive to participate in passing
The fermentation of system food, by natural selection, specific bacterial strain becomes critical function microorganism, these microorganisms shallow lake rich in
The enzyme systems such as powder enzyme, protease and lipase make the various composition of soybean change during the fermentation, form shape, wind
The all more satisfactory product of taste, eating method and healthcare function.
The fermentation process of this patent design is that segmented mixed fungus fermentation can be made due to the mutual promoting action of multi-cultur es
Final product obtains higher probiotics quantity and microbial activity, the benefit materials generated also above the food that single bacterium is fermented,
And mixed fungus fermentation have be not easy to contaminate the high feature of miscellaneous bacteria, stability so that fermenting and producing is more simple and easy to do in technology controlling and process.
Summary of the invention:
The present invention is directed to the crowds for function of intestinal canal disorder to provide a kind of probiotics fermention functional food, and be correlation
The production of product provides Research Thinking.
To achieve the goals above, the present invention provides a kind of solid state fermentation food, is using probiotics to soybean, almond,
Fructus cannabis, brush-cherry seed carry out what mixed bacterium stepwise fermentation obtained, and specific preparation process is as follows:
(1) soy meal, almond powder, fructus cannabis powder, brush-cherry seed powder are chosen, the soy meal of every 6-10g adds almond powder, fire
Edestan powder, each 0-5g of brush-cherry seed powder, obtain mixed raw material, and mixture: water=1:1.25-2 (mass volume ratio) is mixed to prepare
Solid-state fermentation culture medium；
(2) it sterilizes to solid-state fermentation culture medium, condition is 121 DEG C, 20min；
(3) actication of culture: by bacillus subtilis, animal bifidobacteria, bifidobacterium adolescentis, bifidobacterium longum, plant
Lactobacillus, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, Pediococcus acidilactici glycerol tube press 1% (v/v respectively
The volume ratio of activation medium) it is inoculated in activation medium and is activated, pH=7.2-7.4, controlled at 37 DEG C；It lives respectively
After changing 20h, re-activation is carried out by the bacterium amount activation medium of transferring again that connects of 10% (v/v), 37 DEG C of reactivations for 24 hours afterwards must two
Grade seed liquor；
(4) mixed culture solid state fermentation: the first stage by the bacillus subtilis seed liquor in step (3) according to inoculum concentration 1 ×
105~5 × 107CFU/g accesses solid-state fermenter and carries out aerobic standing for fermentation, at 36-38 DEG C after solid state fermentation 12-24h, second
Stage is inoculated with bacillus coagulans, lactobacillus plantarum, animal bifidobacteria, lactobacillus paracasei, bifidobacterium adolescentis, Luo Yishi
Lactobacillus, bifidobacterium longum, eight kinds of strains of Pediococcus acidilactici Mixed Microbes (mixed proportion is equivalent, the access amount of each single bacterium
It is 106-107CFU/g it) accesses in fermentor, anaerobic stands solid state fermentation 12-24h up to probiotics fermention beans at 36-38 DEG C
Solid-state food；
Can directly pack after fermentation it is edible, can also vacuum freeze drying, mouthfeel more preferably health food is made.
The soybean, almond fiber crops, fructus cannabis, brush-cherry seed are commercially available；
Preferably, the mixture: water=1:1.5；
Preferably, the inoculum concentration of bacillus subtilis is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of bacillus coagulans is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of animal bifidobacteria is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of bifidobacterium adolescentis is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of bifidobacterium longum is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of lactobacillus plantarum is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of lactobacillus paracasei is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of Pediococcus acidilactici is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the inoculum concentration of Roy scholar's lactobacillus is 5 × 10 in the solid state fermentation7CFU/g；
Preferably, the solid state fermentation vacuum freeze drying condition vacuum freeze drying -26h for 24 hours, until water content is not more than
10%；
Preferably, the lactobacillus plantarum be lactobacillus plantarum (Lactobacillus plantarum) IOB602, in
On June 29th, 2018 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: Beijing's southern exposure
No. 3 Institute of Microorganism, Academia Sinica, institute of area North Star West Road 1, postcode 100101, deposit number are CGMCC No.16021；
Preferably, the animal bifidobacteria is animal bifidobacteria (Bifidobacterium animalis)
IOB402 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, ground on June 29th, 2018
Location: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode 100101, deposit number are
CGMCC No.16028；
Preferably, the lactobacillus paracasei is lactobacillus paracasei (Lactobacillus paracasei) IOB413,
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 29th, 2018, address: Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number CGMCC
No.16022；
Preferably, the bifidobacterium adolescentis is bifidobacterium adolescentis (Bifidobacterium adolescentis)
IOB506 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, ground on June 29th, 2018
Location: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number are
CGMCC No.16026；
Preferably, the bacillus coagulans be bacillus coagulans (Bacillus coagulans) IOB502, in
On June 29th, 2018 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are as follows: court, Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number are as follows: CGMCC
No.16025；
Preferably, the bacillus subtilis be bacillus subtilis (Bacillus subtilis) IOB430, in
On June 29th, 2018 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are as follows: court, Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number CGMCC
No.16024；
Preferably, the lactobacillus reuteri is lactobacillus reuteri (Lactobacillus reuteri) IOB423,
China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are preserved on June 29th, 2018 are as follows: Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number CGMCC
No.16023；
Preferably, the bifidobacterium longum be bifidobacterium longum (Bifidobacterium longum) IOB515, in
On June 29th, 2018 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are as follows: court, Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number CGMCC
No.16027；
Preferably, the Pediococcus acidilactici is Pediococcus acidilactici (pediococcus acidilactici) IOB701,
China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are preserved on July 10th, 2018 are as follows: Beijing
The institute 3 of Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number CGMCC
No.16077。
The utility model has the advantages that
With the migration of the change of dietary structure, the accelerating rhythm of life and social values, modern is come
It says, close trace it to its cause as FAQs of constipation may be summarized to be at 3 points: first is that water shortage；Second is that gastric dynamic dysfunction；Third is that gastrointestinal tract
In have obstruction.The present invention is exactly directed to three cases above, and problem above is solved in terms of three kinds different.
Firstly, we have selected the strain bacillus coagulans with water-retaining property as one of main fermentation strain, condensation
Bacillus S-layer proteins have water-retaining property, can attract hydrone by charge effect, lock moisture, relax bowel and defecation is only
One has the probiotics of water conservation purging and intestine lubricating action.In addition to this bacillus coagulans can secrete antibacterial coaguin, inhibit harmful bacteria
Breeding promotes beneficial bacterium collaboration proliferation, and fast quick-recovery intestinal flora balance eliminates enterotoxin, releases enteroparalysis, restores enterocinesia
Power.
Secondly, the present invention selects main material of the soybean as fermentation substrate, simultaneous selection, which has, drops dry, ease constipation, defaecation
Brush-cherry seed, fructus cannabis and almond as auxiliary material.The selection criteria of fermentation substrate is the substance using integration of drinking and medicinal herbs.
Finally, being segmented solid-state fermentation technology using the means of fermentation, and using autonomous original a variety of probiotics.
The present invention combine soybean, almond, fructus cannabis, brush-cherry seed each substance characteristics, produced after multi-cultur es stepwise fermentation
The substances such as raw benzoic acid, bacteriocin, and the ingredients such as organic acid, beta galactose glycosides, oligosaccharide dramatically increase, for inhibiting pod membrane
The growth of the harmful bacterias such as bacillus and Escherichia coli, promotes the wriggling of enteron aisle, and relax bowel and defecation prevents constipation from playing important function.
Specific embodiment:
The technical solution of the present invention will be described in detail below with reference to the embodiments and experimental examples, specific embodiment described herein
Be only used to explain the present invention, but the present invention is not limited to following specific embodiments and protection scope it is without being limited thereto.
The percentage sign " % " being related in embodiment refers to mass percent if not specified, and the percentage of solution refers to
Containing the grams of solute in 100mL, percentage between liquid refers to the volume ratio of the solution at 25 DEG C.
Following embodiment will be with lactobacillus plantarum (Lactobacillus plantarum) IOB602, deposit number
CGMCC No.16021；Animal bifidobacteria (Bifidobacterium animalis) IOB402, deposit number CGMCC
No.16028；Lactobacillus paracasei (Lactobacillus paracasei) IOB413, deposit number CGMCC
No.16022；Bifidobacterium adolescentis (Bifidobacterium adolescentis) IOB506, deposit number CGMCC
No.16026；Bacillus coagulans (Bacillus coagulans) IOB502, deposit number are as follows: CGMCC No.16025；It is withered
Careless bacillus (Bacillus subtilis) IOB430, deposit number are CGMCC No.16024；Lactobacillus reuteri
(Lactobacillus reuteri) IOB423, deposit number are CGMCC No.16023；Bifidobacterium longum
(Bifidobacterium longum) IOB515, deposit number are CGMCC No.16027；Pediococcus acidilactici
(pediococcus acidilactici) IOB1010, deposit number solve the present invention for being CGMCC No.16077
Release explanation.
Actication of culture mode used by following embodiment is if not otherwise specified, as follows:
(1) it prepares activation medium: adding 10g soy meal in every 100ml water, obtain activation medium；To activation medium
It sterilizes, condition is 121 DEG C, 25min；
(2) actication of culture: lactobacillus reuteri, bifidobacterium longum, Pediococcus acidilactici, lactobacillus plantarum, animal bifid bar
Bacterium, lactobacillus paracasei, bifidobacterium adolescentis, bacillus coagulans and bacillus subtilis glycerol tube press 1% (v/v respectively
The volume ratio of activation medium) it is inoculated in activation medium and is activated, pH is naturally, controlled at 37 DEG C；20h is activated respectively
Afterwards, by 10% (v/v) connect bacterium amount transfer again activation medium carry out re-activation, reactivation for 24 hours afterwards secondary seed solution.
1 nine kinds of probiotics bacterial mixed solid fermentation bean food techniques of embodiment
1, in raw material soybean and other ratio of adjuvant determination
(1) it dries after being respectively washed soybean, almond, fructus cannabis, brush-cherry seed, is sieved after milling；
(2) it prepares activation medium: adding 10g soy meal in every 100ml water；It sterilizes to activation medium, condition
For, 121 DEG C, 20min；
(3) fermentation medium: the soy meal of every 6-10g is prepared, addition almond powder, fructus cannabis powder, brush-cherry seed powder add 0-
5g；Material-water ratio is 1:1.5；
(4) actication of culture: bacillus subtilis, animal bifidobacteria, bifidobacterium adolescentis, bifidobacterium longum, plant cream
Bacillus, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, Pediococcus acidilactici glycerol tube be inoculated in respectively by 1%
Activation medium is activated, pH=7.2-7.4, controlled at 37 DEG C；After activating 20h respectively, bacterium is connect by 10% (v/v)
Switching activation medium carries out re-activation to amount again, and reactivation obtains secondary seed solution afterwards for 24 hours；
(5) probiotics solid state fermentation: by second level bacillus subtilis seed liquor according to 1 × 106The inoculum concentration of CFU/g accesses
Aerobic fermentation is carried out in 6T solid-state fermenter, after fermenting for 24 hours at 37 DEG C according to inoculum concentration be 1 × 106CFU/g is respectively connected to animal
Bifidobacterium, bifidobacterium adolescentis, bifidobacterium longum, lactobacillus plantarum, lactobacillus paracasei, lactobacillus reuteri, condensation bud
Spore bacillus, Pediococcus acidilactici second level seed mixture liquid carry out secondary inoculation, at 37 DEG C anaerobic fermentation for 24 hours, respectively to fermentation liquid
In number of live bacteria of probiotics detected, as a result such as table 1:
Strain label: No. 1 be bacillus subtilis single bacterium, No. 2 be animal bifidobacteria, No. 3 be bifidobacterium adolescentis, 4
Number be bifidobacterium longum, No. 5 be lactobacillus plantarum, No. 6 be lactobacillus paracasei, No. 7 be lactobacillus reuteri, No. 8 be condensation
Bacillus, No. 9 be Pediococcus acidilactici.
Each group is indicated with label:
Group unification: 8g soy meal+3g almond powder+1g brush-cherry seed powder；
Combine two: 8g soy meal+3g almond powder+5g fructus cannabis powder；
Combine three: 8g soy meal+5g fructus cannabis powder+1g brush-cherry seed powder；
Combine four: 8g soy meal+3g almond powder+5g fructus cannabis powder+1g brush-cherry seed powder.
Then optimal proportion assistant ingredients obtained carries out mixed fermentation, preferably out between soybean and three kinds of auxiliary materials
Optimal proportion.It the results are shown in Table 1.
1 soy meal of table and multiple auxiliary materials mixed fermentation (CFU/g)
As shown in Table 1, solid state fermentation soy meal and almond powder, fructus cannabis powder, proportion optimal between brush-cherry seed powder are big
Bean powder: almond powder: fructus cannabis powder: brush-cherry seed powder=8:3:5:1, the highest viable count obtained are 8.96 × 109CFU/g。
2, the determination of each strain inoculum concentration
The configuration of fermentation medium: soy meal: almond powder: fructus cannabis powder: brush-cherry seed powder=8:3:5:1, fermentation substrate: water
=1:1.5.
In addition to connecing bacterium amount difference, other conditions are the same as embodiment 1-1.
Bacterium amount is connect to be respectively as follows:
Group unification bacillus subtilis 1 × 106CFU/g, the mixed bacterium difference 1 × 10 of eight kinds of bacterium6CFU/g；
Combine two, bacillus subtilises 1 × 106CFU/g, the mixed bacterium difference 5 × 10 of eight kinds of bacterium6CFU/g；
Combine three, bacillus subtilises 5 × 106CFU/g, the mixed bacterium difference 1 × 10 of eight kinds of bacterium6CFU/g；
Combine four, bacillus subtilises 5 × 106CFU/g, the mixed bacterium difference 5 × 10 of eight kinds of bacterium6CFU/g；
Combine five, bacillus subtilises 1 × 107CFU/g, the mixed bacterium difference 1 × 10 of eight kinds of bacterium7CFU/g；
Combine six, bacillus subtilises 1 × 107CFU/g, the mixed bacterium difference 5 × 10 of eight kinds of bacterium7CFU/g；
Combine seven, bacillus subtilises 5 × 107CFU/g, the mixed bacterium difference 1 × 10 of eight kinds of bacterium7CFU/g；
Combine eight, bacillus subtilises 5 × 107CFU/g, the mixed bacterium difference 5 × 10 of eight kinds of bacterium7CFU/g；
After fermentation, the total viable count of probiotics bacterial in gained probiotics solid-state food is detected.
Viable count (CFU/g) under 2 different vaccination amount of table
| Bacillus subtilis viable count | Total viable count of the mixed bacterium of eight kinds of bacterium | Total bacteria count | |
| Group unification | 3.47×108 | 5.45×108 | 8.92×108 |
| Combination two | 3.47×108 | 6.67×108 | 1.01×109 |
| Combination three | 7.68×108 | 7.87×108 | 1.56×109 |
| Combination four | 7.68×108 | 8.79×108 | 1.65×109 |
| Combination five | 8.65×108 | 9.16×108 | 1.78×109 |
| Combination six | 8.65×108 | 9.54×108 | 1.82×109 |
| Combination seven | 9.89×108 | 1.02×109 | 2.0×109 |
| Combination eight | 9.89×108 | 2.12×109 | 3.11×109 |
The results are shown in Table 2, and in the solid-state food of soybean and the fermentation of its auxiliary material, several various combinations ferment, thallus
Inoculum concentration is bacillus subtilis 5 × 107The mixing inoculum concentration of CFU/g, eight kinds of bacterium are 5 × 107When CFU/g, viable count reaches
Highest, although and each combination viable count have by a small margin that difference has all reached higher 9 power, wherein combining eight viable count withered grass
Bacillus has reached 9.89 × 108CFU/g, the mixed bacterium 2.12 × 10 of eight kinds of bacterium9CFU/g。
3, the determination of material-water ratio
The configuration of culture medium in 6T fermentor: soy meal: almond powder: fructus cannabis powder: brush-cherry seed powder=8:3:5:1.
Thallus connects bacterium amount are as follows: bacillus subtilis 5 × 107CFU/g, the inoculum concentration of the mixed bacterium of eight kinds of bacterium respectively 5 ×
107CFU/g。
Soybean and its auxiliary material solid-state fermentation culture medium: the ratio (mass volume ratio) of soybean and its auxiliary material and water is respectively 1:
1.25,1:1.5,1:1.75,1:2.
In addition to conditions above, other conditions are the same as embodiment 1-1.
Number of live bacteria of probiotics in culture medium after fermentation is detected, as a result as seen in Table 3:
When the proportion of 3 soybean of table and its auxiliary material and water is 1:1.25-2, number of live bacteria of probiotics (CFU/g)
| Bacillus subtilis single bacterium viable count | Other eight kinds of total viable count of lactic acid bacteria | Total viable count | |
| 1:1.25 | 5.41×108 | 4.87×108 | 1.03×109 |
| 1:1.5 | 5.79×108 | 4.99×108 | 1.08×109 |
| 1:1.75 | 5.71×108 | 4.76×108 | 1.05×109 |
| 1:2 | 3.87×108 | 3.97×108 | 7.84×108 |
As shown in Table 3, when material-water ratio is 1:1.5, the viable count of bacillus subtilis is 5.79 × 108CFU/g, it is other
The viable count of eight kinds of mixed bacterium is 4.99 × 108CFU/g, total bacteria count have reached highest, and 1.08 × 109CFU/g。
4, the determination of incubation time
6T fermentation tank culture medium are as follows: soy meal: almond powder: fructus cannabis powder: brush-cherry seed powder=8:3:5:1；By mixture with
Water ratio is that 1:1.5 (mass volume ratio) is mixed, 121 DEG C of sterilizing 20min.
Bacillus subtilis and other eight kinds of mixed bacterium are respectively connected to activation medium by inoculum concentration 1% from glycerol tube
Activated, pH naturally, cultivation temperature be 37 DEG C, incubation time 20h, transferred again by 10% inoculum concentration into activation medium
Re-activation is carried out, activation time obtains secondary seed solution afterwards for 24 hours；
Second level bafillus natto seed liquor is pressed 5 × 107Aerobic fermentation is carried out in CFU/g access 6T solid-state fermentation tank,
After fermenting at 37 DEG C, by 5 × 107CFU/g accesses animal bifidobacteria, bifidobacterium adolescentis, bifidobacterium longum, plant cream bar
Bacterium, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, eight kinds of bacterium of Pediococcus acidilactici mixed bacterium (between eight kinds of mixed bacterium
Inoculum concentration be 5 × 107CFU/g secondary seed solution) enters solid-state fermenter, anaerobic fermentation at 37 DEG C.After fermentation,
Obtain viable type fermentation bean food.
Fermentation time is 1. bacillus subtilis 12h, the fermentation time 12h of other eight kinds of bacterium；
2. bacillus subtilis 12h, the fermentation time of other eight kinds of bacterium is for 24 hours；
3. bacillus subtilis is for 24 hours, the fermentation time 12h of other eight kinds of bacterium；
4. bacillus subtilis is for 24 hours, the fermentation time of other eight kinds of bacterium is for 24 hours.
After fermentation, count plate is carried out to the product after solid state fermentation respectively.
The viable count of lactic acid bacteria in 4 solid state fermentation bean food of table
| 1 | 2 | 3 | 4 | |
| Bacillus subtilis | 5.45×108 | 6.69×108 | 5.73×108 | 6.24×108 |
| Eight kinds of mixed bacterium | 4.49×108 | 4.1×108 | 4.36×108 | 5.24×108 |
| Total bacteria count | 5.16×109 | 6.26×109 | 5.54×109 | 5.01×109 |
Fermentation results show, when the other eight kinds of bacterium of bacillus subtilis 12h fermentation time for 24 hours when, fermentation results
Reach best, total bacteria count has reached 6.26 × 109CFU/g。
5, the measurement of ingredient after fermenting
Other conditions are the same as embodiment 1-1, bafillus natto, animal bifidobacteria, bifidobacterium adolescentis, long bifid bar
Inoculum concentration is when bacterium, lactobacillus plantarum, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, Pediococcus acidilactici fermentation
5×107CFU/g, by soy meal: almond powder: fructus cannabis powder: brush-cherry seed powder=8:3:5:1 configures fermentation medium, fermented and cultured
The material-water ratio of base is 1:1.5；Fermentation time is bacillus subtilis 12h, and the fermentation time of other eight kinds of bacterium is for 24 hours.
The detection method of lipase is using standard GB/T/T 5523-2008 grain and oil detection grain, the lipase activity of oil plant
The measurement of dynamic degree；The detection method of protease is using national current standard GB 5009.5-2010 national food safety standard food
The measurement of middle protein；The detection method of amylase is national current standard GB/T 24401-2009 alpha Amylase preparation；Fat
The measuring method of grease and benzoic acid is using the variation of gas chromatography-analytical reagent composition fermentation front and back；The measurement of chain fatty acid
Method is using oligopeptide (in food) in beverage, the measuring method of short chain fatty acids and γ-aminobutyric acid；The measuring method of lactic acid
It is examined using standard GB/T 4789.35-2010 national food safety standard food microbiological examination lactic acid bacteria；β-gala
The detection method of glycosidase is divided light using national current standard GB/T 33409-2016 betagalactosidase activity detection method
Degree method；The measuring method of organic acid uses the measurement of GB 5009.157-2016 national food safety standard food organic acid；Liu
The pretty chromatography method measurement mentioned in " detection of a strains of lactic acid bacteria bacteriocin and its research of the optimum condition generated "；It is oligomeric
The measuring method of sugar uses GB/T 22491-2008 soyabean oligosaccharides；The detection method of dietary fiber is using country mark GB
5009.88-2014 the measurement of dietary fiber is quasi- in national food safety standard food.
The active constituent variation of 5 soybean solid-state of table fermentation front and back
| Detect ingredient | Before fermentation | After fermentation |
| Amylase (U/g) | 124.6 | 424.3 |
| Protease (U/g) | 174.4 | 524.4 |
| Lipase (U/g) | 113.2 | 437.6 |
| Benzoic acid (%) | 0 | 0.073 |
| Fatty grease (%) | 42% | 48% |
| Short chain fatty acids (mg/L) | 153.23 | 205.12 |
| Lactic acid (mg/100g) | 52.32 | 145.23 |
| Beta galactosidase (U/mL) | 0.68 | 1.53 |
| Organic acid (mg/100g) | 163.31 | 465.92 |
| Bacteriocin (Da) | 0 | 2300 |
| Oligosaccharide (%) | 9% | 45% |
| Dietary fiber (g/100g) | 2.43 | 6.41 |
Caused by constipation is large intestine mechanical movement, shows as difficult defecation, discomfort, rectum and splash swollen and incomplete emptying, row
The empty time extends.The cause for influencing defecation process and constipation occurring is known as very much, most important of them first is that dietary factor,
The non-starch polysaccharide for increasing to large intestine, the normal flora for restoring enteron aisle facilitate relief of constipation, the cream after a large amount of intake fermentations
Sour bacterium can promote intestines peristalsis, relief of constipation.
The probiotics such as Bacillus acidi lactici and Bifidobacterium in enteron aisle can decompose organic acid (such as lactic acid generated in reference state
Deng), make to form low ph environment in enteron aisle, inhibit enteric pathogenic bacteria growth and breeding, promote beneficial bacterium growth and breeding, increase has
The quantity of beneficial bacteria makes beneficial bacterium advantage occupy-place, to establish good microbiota.In addition, organic acid can increase the compacted of intestines
Dynamic frequency, reinforces the secretion of enteron aisle, while promoting digestion and absorption of the enteron aisle to nutriment.Product after everfermentation, fat
The content of grease has micro raising, can effectively improve intestinal contents to the lubricating action of intestinal mucosa, while solid product exists
Benzoic acid is generated by the effect of enzyme in alimentary canal, the activity of pepsin can be inhibited, so product has relax bowel and defecation
Effect.
Lactic acid intolerance is to lack beta galactosidase due to mucous membrane of small intestine to cause lactic acid digestion insufficient, into large intestine
Lactic acid degraded by bacterial enzyme, cause gas in intestinal tube to increase, it is a series of based on flatulence, abdominal pain and osmotic diarrhea etc.
Clinical symptoms.Raw material in the present invention beta galactosidase after everfermentation dramatically increases, and flatulence etc. can be effectively relieved
Symptom.
In addition, above-mentioned data illustrate to ferment by multi-cultur es segmentation mix of the invention, the oligosaccharide content in product becomes
Change significant.Oligosaccharide is nutriment necessary to bifidobacterium growth, and Bifidobacterium generates acetic acid and lactic acid using oligosaccharide
Equal metabolites, can inhibit the growth of the harmful bacterias such as Bacillus perfringens and Escherichia coli, promote the wriggling of enteron aisle, prevent just
It is secret.Dietary fiber in human oral cavity, stomach, small intestine, be not digested absorption, to increase the volume of intestinal contents, maintain enteron aisle
Interior internal pressure makes fluffy of defecating, and enteron aisle is easier to push swill, effectively removes waste and rubbish in enteron aisle fold, will
Waste after nutrient absorption is removed, to play promotion, often band is wriggled.
It in summary it can be seen, the beans solid-state food after a variety of probiotic mixed fermentations combines the spy of various substances
Property.Product after prebiotic strain fermentation contains various digestive ferments, improves alimentary canal physiology, generates creatine, reduces intestinal pH etc..
Certain micro-organisms can also form the chelate for being easy to be absorbed and utilized with some mineral elements, promote its absorption, improve animal to micro-
The utilization efficiency of secondary element.
The research of 2 fermentation soybean of embodiment and auxiliary material to mouse relax bowel and defecation healthcare function
Prepare clean grade mouse 12, as blank control group；Mouse with Constipation Model 60, is randomly divided into 5 groups,
I.e. white group of constipation Model Space, Constipation Model control group, Constipation Model experimental group distinguish the high, medium and low dosage group of feeding soy food product.
After mouse fasting 12h, blank control group, Constipation Model group give isometric(al) physiological saline, and Constipation Model control group gives Mai Jin
Sharp board probiotic powder, once a day, one bag (1.5g) every time；Embodiment 1-5 preparation is given in stomach-filling to remaining groups of animals respectively
Tempeh (product for first obtaining embodiment 1-5 is through vacuum freeze drying to water content 12%) 15g/ times, 10g/ times,
5g/ times, twice daily.Mouse single cage is raised after stomach-filling, normal diet water inlet, when record observes every small mouse's head grain defecation
Between, defecation weight in defecation grain number and 6h in 6h.
Influence of 6 soy food product of table to mice with constipation
| Dosage | Number of animals/only | First just time (min) | Fecal grains (grain) | Stool weight (g) |
| 15g//times | 12 | 75±25.5 | 28.2±5.76 | 0.58±0.32 |
| 10g//times | 12 | 110±20 | 24.6±9.87 | 0.43±0.16 |
| 5g//times | 12 | 185±35 | 23.5±5.87 | 0.38±0.32 |
| Constipation Model control group | 12 | 204±32 | 20.8±12.2 | 0.32±0.156 |
| Constipation Model blank group | 12 | 265±19 | 15.2±8.37 | 0.25±0.21 |
| Blank control group | 12 | 42±38 | 32.8±3.87 | 0.62±0.26 |
As shown in Table 6, after to mice with constipation consumable products, the first just time is obviously shortened, and fecal grains dramatically increase, excrement
Just weight obviously increases.Low dose group just has more apparent symptom, and the product of middle and high dosage has a significant effect.If
Excrement residence time in enteron aisle is too long, and the moisture that will lead in excrement is excessively absorbed by enteron aisle, and excrement will become
In stiff, so this situation can be improved by shortening defecation time.All substances that can be improved excrement moisture holding capacity are pointed out in research
It is logical that the substance (various water insoluble dietary fibers) of (various water-soluble dietary fibers) or promotion intestines peristalsis can play ease constipation
Just effect.
3 fermented soybean product of embodiment treats anti-constipation effect analysis
Choosing 58 has constipation aspect Disease as research object.It is divided into two groups, every group each 29.It is right
According to daily 3 trimebutine maleate dispersion tablets are organized, each taking 1.Observation group uses the fermented soybean of embodiment 1-5
Product, 3 times a day, each 5g.The course for the treatment of is 4 weeks.
In the treatment of constipation type patient, western medicine is most commonly seen therapeutic scheme, Trimebutine Maleate dispersion
Piece can effectively inhibit and the gastrointestinal function of excited patient, thus will appear the effect of bidirectional modulation, improves patient's stomach and intestine
The micro-ecological environment in road, improvement related indication for patient is advantageously.MOT is a kind of gastrointestinal hormone of isolated polypeptide,
Facilitation is played to gastrointestinal tract decomposition electrolyte, Spinal Cord Oedema, accelerates the movement of gastrointestinal tract.VIP is then base polypeptide, can not only
Adjust neurons secrete, moreover it is possible to promote the peristaltic contraction of enteron aisle.SS can inhibit the synthesis of gastric acid and the movement of gastrointestinal tract, stomach and intestine
The secretion of road hormone, has the function that relief of constipation.From data it can also be seen that the therapeutic effect of observation group better than pair
According to group, VIP, SS index level are decreased obviously, and MOT index level obviously rises, and are compared between group, and difference has statistical significance.
See Table 7 for details.
7 two groups of patients blood plasma's gastrointestinal hormone levels of table compare (x ± s, pg/ml)
It can be seen that fermented soybean food is improved therapeutic effect for treatment constipation type patient, relief of constipation symptom is adjusted
It is horizontal to save blood plasma gastrointestinal hormone, therapeutic process is safe and effective, which can be adopted.
Claims (5)
1. a kind of probiotics fermention functional food, which is characterized in that be using probiotics to soybean, almond, fructus cannabis, Yu Li
Benevolence carries out what mixed bacterium stepwise fermentation obtained, and the probiotics is bacillus subtilis, animal bifidobacteria, youth bifid bar
Bacterium, bifidobacterium longum, lactobacillus plantarum, lactobacillus paracasei, lactobacillus reuteri, bacillus coagulans, Pediococcus acidilactici.
2. a kind of probiotics fermention functional food as described in claim 1, which is characterized in that the mixed bacterium stepwise fermentation tool
Body is as follows: the first stage is by bacillus subtilis seed liquor according to inoculum concentration 1 × 105~5 × 107CFU/g accesses solid state fermentation training
It supports base and carries out aerobic standing for fermentation, at 36-38 DEG C after solid state fermentation 12-24h, second stage is inoculated with bacillus coagulans, plant
Lactobacillus, animal bifidobacteria, lactobacillus paracasei, bifidobacterium adolescentis, lactobacillus reuteri, bifidobacterium longum, lactic acid sheet
The Mixed Microbes of eight kinds of strains of coccus, mixed proportion are equivalent, and the access amount of each single bacterium is 106-107Nothing at CFU/g, 36-38 DEG C
Oxygen stands solid state fermentation 12-24h up to probiotics fermention beans solid-state food.
3. a kind of probiotics fermention functional food as claimed in claim 2, which is characterized in that the solid-state fermentation culture medium
It is to add almond powder, fructus cannabis powder, each 0-5g of brush-cherry seed powder in the soy meal of every 6-10g, obtain mixed raw material, will mix
Material: water=1:1.25-2 is mixed to prepare solid-state fermentation culture medium；121 DEG C, 20min sterilizing obtains.
4. a kind of probiotics fermention functional food as claimed in claim 1 or 2, which is characterized in that
The lactobacillus plantarum is lactobacillus plantarum (Lactobacillus plantarum) IOB602, deposit number CGMCC
No.16021；
The animal bifidobacteria is animal bifidobacteria (Bifidobacterium animalis) IOB402, and deposit number is
CGMCC No.16028；
The lactobacillus paracasei is lactobacillus paracasei (Lactobacillus paracasei) IOB413, and deposit number is
CGMCC No.16022；
The bifidobacterium adolescentis is bifidobacterium adolescentis (Bifidobacterium adolescentis) IOB506, and preservation is compiled
Number be CGMCC No.16026；
The bacillus coagulans are bacillus coagulans (Bacillus coagulans) IOB502, deposit number are as follows: CGMCC
No.16025；
The bacillus subtilis is bacillus subtilis (Bacillus subtilis) IOB430, deposit number CGMCC
No.16024；
The lactobacillus reuteri is lactobacillus reuteri (Lactobacillus reuteri) IOB423, and deposit number is
CGMCC No.16023；
The bifidobacterium longum is bifidobacterium longum (Bifidobacterium longum) IOB515, deposit number CGMCC
No.16027；
The Pediococcus acidilactici is Pediococcus acidilactici (Pediococcus acidilactici) IOB701, and deposit number is
CGMCC No.16077。
5. a kind of probiotics fermention functional food as claimed in claim 1 or 2, which is characterized in that the preparation method is as follows:
(1) soy meal, almond powder, fructus cannabis powder, brush-cherry seed powder are chosen, soy meal: almond powder: fructus cannabis powder: brush-cherry seed powder=
8:3:5:1 obtains mixed raw material, and mixture: water=1:1.5 is mixed to prepare solid-state fermentation culture medium；121 DEG C, sterilizing
20min；
(2) mixed culture solid state fermentation: the first stage is by bacillus subtilis seed liquor according to inoculum concentration 5 × 107CFU/g accesses solid-state
Fermentor carries out aerobic standing for fermentation, and at 37 DEG C after solid state fermentation 12h, second stage is inoculated with bacillus coagulans, plant cream bar
Bacterium, animal bifidobacteria, lactobacillus paracasei, bifidobacterium adolescentis, lactobacillus reuteri, bifidobacterium longum, Pediococcus acidilactici
The Mixed Microbes of eight kinds of strains, the access amount of each single bacterium are 5 × 107CFU/g, anaerobic stands solid state fermentation for 24 hours at 37 DEG C to obtain the final product
Probiotics fermention beans solid-state food；
Edible or vacuum freeze drying is directly packed after fermentation, and health food is made.
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| CN111011493A (en) * | 2019-12-24 | 2020-04-17 | 广东燕塘乳业股份有限公司 | Preparation method of solidified fermented milk rich in animal bifidobacterium and fermented milk prepared by preparation method |
| CN114375977A (en) * | 2020-10-20 | 2022-04-22 | 汉义生物科技（北京）有限公司 | Fructus cannabis oligopeptide and preparation method thereof |
| CN115500493A (en) * | 2022-09-19 | 2022-12-23 | 湖南吻野现代农业科技有限公司 | Probiotic peanut and preparation method thereof |
| CN116731937A (en) * | 2023-08-15 | 2023-09-12 | 天津创源生物技术有限公司 | Preparation method and application of Wittman coagulans IOB502 zymocyte powder |
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| CN117511825A (en) * | 2023-12-29 | 2024-02-06 | 天津创源生物技术有限公司 | Application of segment fermentation inoculant of Wittman coagulans IOB502 in alcoholic liver injury |
| CN117511825B (en) * | 2023-12-29 | 2024-04-05 | 天津创源生物技术有限公司 | Application of segment fermentation inoculant of Wittman coagulans IOB502 in alcoholic liver injury |
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