CN105942216A - Probiotics fermented five bean food and preparation method thereof - Google Patents
Probiotics fermented five bean food and preparation method thereof Download PDFInfo
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- 244000046052 Phaseolus vulgaris Species 0.000 title claims abstract description 67
- 235000010627 Phaseolus vulgaris Nutrition 0.000 title claims abstract description 67
- 235000013305 food Nutrition 0.000 title claims abstract description 45
- 239000006041 probiotic Substances 0.000 title claims abstract description 40
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000000855 fermentation Methods 0.000 claims abstract description 92
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- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 46
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Classifications
-
- 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
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- 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
Abstract
The present invention provides a probiotics fermented five bean food and a preparation method thereof, belonging to the technical field of fermentation. The probiotics fermented five bean food is obtained by using five beans as fermentation substrates and using at least two of bacillus subtilis, lactobacillus plantarum and bifidobacterium animalis to conduct mixed bacterium liquid or solid fermentation. The obtained products are high in lactic acid bacterium contents, the contents of daidzein, genistein, total phenols, gamma-aminobutyric acid and other active ingredients are increased significantly, so that the probiotics fermented five bean food can promote digestion, regulate intestinal tracts, and improve human body immunity. Besides, the five beans are mixed to nourish five organs, and harmonize and regulate the body at the same time.
Description
Technical field:
The invention belongs to fermentable application, relate to a kind of probiotics fermention five elements' bean liquid beverage with solid
State food and preparation method thereof.
Background technology:
Along with social progress and the raising of living standards of the people. the demand of nutrition is met by people beyond simple
Existence or prevent the category of deficiency disease, and also serve as the important means of disease preventing and treating.Reasonably diet structure,
It is that the mankind improve life quality, extend the existence time limit, the effective ways of state of keeping fit.Bean food belongs to
The food that grain rice and aqueous phase melt, is suitable for four seasons, the safest but also nutrition.Described in the traditional Chinese medical science: " five colors enter the five internal organs,
Equal six abdomens of the five elements ", illustrate that beans has the effect of balance the five internal organs, i.e. Semen Phaseoli reinforcing the heart is dirty, and Semen Glycines spleen reinforcing is dirty, green
Bean tonifying liver is dirty, and Semen Vignae Cylindricae tonifying the lung is dirty, and Semen sojae atricolor the kidney invigorating is dirty.The vital organs of the human body have been taken into account by five elements' bean, the coolest the driest,
Rush down not imapirment of the spleen and stomach, mend and do not increase stasis, longevity of preserving youthful looks.
Probiotic bacteria is the active microorganism that a class is useful to host, is colonizated in host intestine, reproductive system, energy
Produce definite health efficacy thus improve host's microecological balance, the active beneficial microorganism of performance beneficial effect
General name.Probiotic bacteria application in food is as the frontier of biological study, because its goods can regulate human body intestinal
Road colony balance, has important function to healthy, thus is increasingly becoming the research focus of functional food.
Beverage that probiotics fermention five elements' bean produces and food is utilized to have good to human body intestinal canal microorganism species
Regulation effect, this product not only remains some active component of material bean apoplexy due to endogenous wind, as polysaccharide, dietary fiber,
Bioflavonoids etc. have the material of health-care effect to body, and the most also can produce special flavor thing
Matter, improves the taste of product on the basis of the original nutritional labeling of retained product so that it is mouthfeel more preferably, be more easy to by
Absorption of human body, and facilitating digestion, conditioning intestinal while nourish the five internal organs: i.e. Semen sojae atricolor the kidney invigorating, strengthen kidney it
Gas, keep healthy skin care；Semen Glycines enhances metabolism, and improves spleen effect；Semen Vignae Cylindricae lung moistening, QI invigorating in reason；Semen Phaseoli
Reinforcing the heart, purify the blood logical blood；Semen phaseoli radiati removing liver heat and toxic substances, the five internal organs are with mending, and harmony is nursed one's health.
Summary of the invention:
It is an object of the invention to utilize multiple probiotic bacteria to mix bacterium and five elements' bean is carried out a kind of probiotic bacteria of fermentation acquisition
Functional food.
A kind of probiotics fermention five elements' bean food, is as fermentation substrate using five elements' bean, use bacillus subtilis,
After at least two in Lactobacillus plantarum and animal bifidobacteria carries out mixed bacterium solution state or solid fermentation, preparation and
?.
Described five elements' bean is Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines；
Preferably, described Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillus plantarum) TK9, this bacterium
Strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms in December in 2015 on the 17th
The heart, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode
100101, deposit number is CGMCC No.11891；
Preferably, described animal bifidobacteria is animal bifidobacteria (Bifidobacterium animalis) 937,
This bacterial strain is preserved in the common micro-life of China Committee for Culture Collection of Microorganisms on the 17th in December in 2015
Thing center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postal
Compiling 100101, deposit number is CGMCC No.11892；
Preferably, described bacillus subtilis is bacillus subtilis (Bacillus subtilis) TK-1, preservation
Numbered CGMCC No.4731.
The preparation method of above-mentioned probiotics fermention five elements' bean food, specifically comprises the following steps that
(1) actication of culture: bacillus subtilis, Lactobacillus plantarum and animal bifidobacteria go out from glycerol pipe respectively
Sending out, access activation medium by inoculum concentration 1% and once activate, pH is natural, and cultivation temperature is 36-38 DEG C,
Incubation time 18-20h, as primary seed solution；Cultivation accesses identical activation training by 10% inoculum concentration after completing
Support in base, in 36-38 DEG C, again activate 20-24h under pH natural conditions, as secondary seed solution；
(2) liquid fermentation is cultivated: strain is seeded to from secondary seed solution liquid fermentation culture medium respectively and carries out
Any two bacterium or the mixed fermentation of three kinds of bacterium, 36-38 DEG C, mixed bacterium solution state standing for fermentation 12～24 under pH natural conditions
h；
After acid adjustment, perfumery, homogenizing, sterilization, five elements' bean liquid fermentation beverage it is after fermentation ends；
(3) solid state fermentation culture: the secondary seed solution in step (1) is accessed in solid-state fermentation culture medium and carries out
Any two bacterium and the mixed fermentation of three kinds of bacterium, at 36-38 DEG C, mixed culture solid state fermentation 24-48h i.e. obtains five elements' bean benefit
Raw bacterium solid fermentation food；After fermentation ends, bean or pea can directly be packed edible, it is possible to preserves after vacuum lyophilization；
Described activation medium is: after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, often
100mL water adds the bean or pea making beating homogenizing after 5g mixed in equal amounts, homogenizing temperature 65 DEG C, pressure 20.0MPa；
Described liquid fermentation culture medium is: after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned
Pulverizing, after crossing 100 mesh sieves, by the addition of 5-10% (mass volume ratio), mixs homogeneously with water, 121 DEG C
Sterilizing 20min；
In described liquid fermentation, the inoculum concentration of Lactobacillus plantarum and animal bifidobacteria is 106-107CFU/ml,
The inoculum concentration of bacillus subtilis is 104-105CFU/ml；
Preferably, the inoculum concentration of described Lactobacillus plantarum and animal bifidobacteria is 5 × 107CFU/ml, hay
The inoculum concentration of bacillus cereus is 5 × 104CFU/ml；
Preferably, the temperature 37 DEG C that described liquid fermentation is cultivated；
Preferably, described mixed culture liquid state fermentation time 24h；
Preferably, the processing condition after described liquid fermentation terminates is homogenizing temperature 60 DEG C-70 DEG C, pressure
15.0-20.0MPa；
Preferably, it is 90 DEG C that described liquid fermentation terminates rear sterilization conditions, 10min；
Preferably, described five elements' bean liquid fermentation beverage, water content is not more than 95%；
Described solid-state fermentation culture medium is: after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned,
Mix by material-water ratio 1:1-1.5 (mass volume ratio), soak 8-10h at 4 DEG C and prepare, 121 DEG C of sterilizings
20min；
In described solid fermentation, the inoculum concentration of Lactobacillus plantarum and animal bifidobacteria is 106-107CFU/g is withered
The inoculum concentration of grass bacillus cereus is 104-105CFU/g；
Preferably, in described solid fermentation, the inoculum concentration of Lactobacillus plantarum and animal bifidobacteria is
1×107CFU/g, the inoculum concentration of bacillus subtilis is 5 × 104CFU/g；
Preferably, the cultivation temperature of described solid fermentation is 37 DEG C；
Preferably, described solid-state mixed fungus fermentation time 48h；
Preferably, described solid fermentation vacuum lyophilization condition is done for carrying out vacuum freezing after-70 DEG C of pre-freeze 6h
Dry 24h, is not more than 5% to water content；
Preferably, described Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillus plantarum) TK9, preservation
Numbered CGMCC No.11891；
Preferably, described animal bifidobacteria is animal bifidobacteria (Bifidobacterium animalis) 937,
Deposit number is CGMCC No.11892；
Preferably, described bacillus subtilis is bacillus subtilis (Bacillus subtilis) TK-1, preservation
Numbered CGMCC No.4731.
Beneficial effect:
1, the present invention uses five kinds of bean or pea mixing making beating homogenizing carry out liquid fermentation and use granulate bean or pea to carry out admittedly
The state fermenting and producing probiotic bacteria beverage based on lactic acid bacteria and food.Lactic acid bacteria or lactic acid bacteria is used to combine hay bud
Spore bacillus is fermented, and can produce the tart flavour small-molecule substance such as lactic acid and acetic acid, and these small-molecule substances give
The improvement of the organoleptic features such as product distinctive acid fragrance, color and luster and mouthfeel and variation.
2, in five elements' bean liquid beverage after probiotics fermention and solid-state food, active component content significantly improves,
As daidzein, genistein etc. can prevent cardiovascular disease, improve osteoporosis and prophylaxis of cancer；Liquid is drunk
Material and solid-state food in total aldehydes matter respectively up to 8.59 (mg GAE/g) and 38.93 (mg GAE/g),
This material has antioxidation, strengthening blood vessel wall, promotion gastrointestinal disturbances, reduction blood fat, increases passive protective physical fitness
Act on and can prevent the formation of arteriosclerosis and thrombosis；γ-aminobutyric acid in liquid beverage and solid-state food
(GABA) respectively reach 2.517mg/g and 6.673mg/g, hypertension is had and alleviates and effect for the treatment of.
3, five kinds of bean or pea mixed fermentation, compares single bean or pea nourishing effects more powerful, combines various bean or pea
Advantage, will Semen Phaseoli blood pressure lowering reinforcing the heart, Semen phaseoli radiati removing toxic substances tonifying liver, Semen Glycines replenishing QI to invigorate the spleen, Semen Vignae Cylindricae help digestion lung moistening and Semen sojae atricolor
Effect of enriching yin and nourishing kidney combines, and the five elements coordinate to complete the conditioning of health natural harmony together.
Specific embodiments
Below in conjunction with detailed description of the invention, the invention will be further described, but the present invention is not limited to following tool
Body examples of implementation.
In following embodiment, activation medium is: by Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts
After cleaning, every 100mL water adds the making beating homogenizing of the bean or pea after 5g mixed in equal amounts, homogenizing temperature 65 DEG C,
Pressure 20.0MPa.
Embodiment 1, probiotic bacteria liquid fermentation researchonthe technology
(1) inoculating proportion is determined
Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 go out from glycerol pipe respectively
Sending out, access activation medium by inoculum concentration 1%, pH is natural, and cultivation temperature is 36 DEG C, and incubation time is 18h,
Once activate, as primary seed solution；From primary seed solution, access activation training by 10% inoculum concentration
Foster base carries out re-activation, and as secondary seed solution, cultivation temperature is 36 DEG C, and pH is natural, and incubation time is
20h。
From secondary seed solution by different the bacterium amount that connects inoculation Lactobacillus plantarum TK9, animal bifidobacteria 937
With any two bacterium in bacillus subtilis TK-1 and simultaneously inoculation three kinds of bacterium to liquid fermentation culture medium (liquid
Fermentation medium is: pulverizing after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, and crosses 100
After mesh sieve, by the addition of 5% (mass volume ratio), mix homogeneously with water, 121 DEG C of sterilizing 20min) in
Ferment.
TK9, the bacterium amount that connects of 937 are respectively: 5 × 106CFU/ml；1×107CFU/ml；5×107CFU/ml；
1×108CFU/ml；The inoculum concentration of bacillus subtilis TK-1 is 5 × 104CFU/ml, pH are natural, cultivate temperature
Spend 36 DEG C, quiescent culture 24h, enters the total viable count of lactic acid bacteria in gained probiotic bacteria liquid beverage after fermentation
Row detection.
The total viable count of lactic acid bacteria in table 1 liquid fermentation beverage
Result is as shown in table 1, Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis TK-1
During three kinds of bacterium mixed fermentation, the total viable count of lactic acid bacteria in fermentation liquid is the highest.The inoculum concentration of bacillus subtilis is
5×104During CFU/ml, the bacterium amount that most preferably connects of two strains of lactic acid bacteria is 5 × 107CFU/ml, i.e. records gained prebiotic
Total viable count in bacterium five elements' bean beverage is 6.55 × 109CFU/ml.Wherein, Lactobacillus plantarum viable count
4.15×109CFU/ml, animal bifidobacteria viable count is 2.40 × 109CFU/ml。
(2) determination of incubation time
Inoculating proportion have been determined as three kinds of bacterium inoculate simultaneously viable count the highest under conditions of, by Lactobacillus plantarum
The glycerol pipe of TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 is inoculated in activation training by 1% respectively
It is natural that foster base once activates 18h, pH, and cultivation temperature is 37 DEG C, transfers into phase by the bacterium amount that connects of 10%
With in re-activation culture medium, carrying out re-activation 20h, pH nature, cultivation temperature is 37 DEG C；In liquid
(after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, pulverizing, crosses 100 mesh to fermentation medium
After sieve, by the addition of 10% (mass volume ratio), mix homogeneously with water, 121 DEG C of sterilizing 20min) in
Three kinds of bacterial strains of inoculation carry out mixed fungus fermentation simultaneously.
The inoculum concentration of Lactobacillus plantarum TK9 and bacillus bifidus 937 is 5 × 107CFU/ml, bacillus subtilis
Inoculum concentration be 5 × 104CFU/ml, under the conditions of cultivation temperature 37 DEG C, pH is natural, respectively quiescent culture
Respectively fermentation liquid is carried out viable lactic acid bacteria counting after 12h, 24h, 36h, 48h.
The total viable count of lactic acid bacteria in table 2 liquid fermentation beverage
Result is: the probiotics fermention five elements' optimal incubation time of bean liquid beverage is 24h, records lactic acid bacteria and always lives
Bacterium number is 6.10 × 109, wherein Lactobacillus plantarum TK9 viable count is 3.80 × 109, animal bifidobacteria 937
Viable count is 2.30 × 109。
(3) active component change before and after fermentation
By the glycerol pipe of Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 respectively
Being inoculated in activation medium cultivation by 1% and once activate 18h, pH nature, cultivation temperature is 37 DEG C,
Transfer in identical re-activation culture medium by the bacterium amount that connects of 10%, carry out re-activation 20h, pH nature,
Cultivation temperature is 37 DEG C, in five elements' bean liquid fermentation culture medium (by Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines
Pulverizing after mixed in equal amounts cleaning, after crossing 100 mesh sieves, by the addition of 5% (mass volume ratio), mixes with water
Close uniformly, 121 DEG C of sterilizing 20min) in inoculate three kinds of bacterial strains simultaneously and carry out mixed fungus fermentation (and with any two kinds of bacterium
The combination of strain and single bacterium fermentation are as comparison).The inoculum concentration of Lactobacillus plantarum TK9 and bacillus bifidus 937 is
5×107CFU/ml, the inoculum concentration of bacillus subtilis TK-1 is 5 × 104CFU/ml, cultivation temperature 37 DEG C,
Under pH natural conditions after quiescent culture 24h, measuring the change of each active component content in fermentation liquid, result is such as
Table 3.
After fermentation ends, the water content of gained five elements bean liquid fermentation beverage is 90%.
Use same method, by Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis
TK-1 replaces with Lactobacillus plantarum CGMCC 1.6971, animal bifidobacteria CGMCC 1.3003 and respectively
Bacillus subtilis CGMCC 1.3358, repeats above-mentioned experiment, result such as table 4.
GB/T 23788-2009 method is utilized to measure isoflavone content；GB/T12143.2-89 method is utilized to survey
Determine the content of amino nitrogen；Utilize Syrups by HPLC GABA content；Utilize forint phenol reaction assay total
Phenols content；Oxygen index method (ORAC) of living is utilized to measure polyphenoils content.
The mensuration of active substance in table 3 liquid fermentation beverage
The mensuration of active substance in table 4 liquid fermentation beverage
2 one kinds of five elements' bean liquid fermentation beverages of embodiment and preparation method thereof
(1) actication of culture: bacillus subtilis TK-1, Lactobacillus plantarum TK9 and animal bifidobacteria 937
Respectively from glycerol pipe, accessing activation medium by inoculum concentration 1% and once activate, pH is natural, cultivates
Temperature is 37 DEG C, and incubation time 20h, as primary seed solution；Cultivation accesses phase by 10% inoculum concentration after completing
With in activation medium, in 37 DEG C, again activate 24h under pH natural conditions, as secondary seed solution；
(2) liquid fermentation is cultivated: strain is seeded to from secondary seed solution liquid fermentation culture medium respectively and carries out
The mixed fermentation of three kinds of bacterium, the inoculum concentration of Lactobacillus plantarum TK9 and bacillus bifidus 937 is 5 × 107CFU/ml,
The inoculum concentration of bacillus subtilis TK-1 is 5 × 104CFU/ml, 37 DEG C, mixed bacterium solution state under pH natural conditions
Standing for fermentation 24h；
Liquid fermentation culture medium is: pulverizing after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned,
After crossing 100 mesh sieves, by the addition of 5% (mass volume ratio), mix homogeneously with water, 121 DEG C of sterilizing 20min；
(3) (mass volume ratio) sucrose carries out acid adjustment perfumery to add 6% after fermentation ends, at homogenizing temperature 70
Degree Celsius, homogenization pressure be to carry out homogenizing under the conditions of 20Mpa, after 90 degrees Celsius of sterilization 10min, be five
Row bean liquid fermentation beverage, water content is 95%.
Embodiment 3 probiotic bacteria solid fermentation food technology
(1) inoculating proportion is determined
Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 go out from glycerol pipe respectively
Sending out, access activation medium by inoculum concentration 1% and once activate, as primary seed solution, cultivation temperature is
36 DEG C, incubation time is that 18h, pH are natural.From primary seed solution, carry out by 10% inoculum concentration inoculation
Re-activation, as secondary seed solution, cultivation temperature is 36 DEG C, and pH is natural, and incubation time is 20h.
From secondary seed solution by different the bacterium amount that connects inoculation Lactobacillus plantarum TK9, animal bifidobacteria 937
With any two bacterium of bacillus subtilis TK-1 and simultaneously three kinds of bacterium of inoculation to solid medium (by Semen Phaseoli, Semen phaseoli radiati,
After Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts are cleaned, mix by material-water ratio 1:1.5 (mass volume ratio), 4 DEG C
Lower immersion 8h, 121 DEG C of sterilizing 20min) in ferment.
TK9, the bacterium amount that connects of 937 are respectively: 5 × 106CFU/g；1×107CFU/g；5×107CFU/g；
1×108CFU/g；The inoculum concentration of bacillus subtilis TK-1 is 5 × 104CFU/g, pH are natural, cultivation temperature
37 DEG C, cultivate 48h, the total viable count of lactic acid bacteria in gained probiotic bacteria solid-state food after fermentation is detected.
The total viable count of lactic acid bacteria in table 5 solid fermentation food
Result is as shown in table 5, Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis mixing
During fermentation, the total viable count of lactic acid bacteria in five elements' bean solid fermentation food is the highest.The inoculum concentration of bacillus subtilis
It is 5 × 104During CFU/g, the bacterium amount that most preferably connects of two strains of lactic acid bacteria is 1 × 107CFU/g.I.e. record probiotic bacteria five
The total viable count of lactic acid bacteria in row bean solid fermentation food is 2.57 × 1010CFU/g.Wherein, Lactobacillus plantarum is lived
Bacterium number 1.87 × 1010CFU/g, the viable count of animal bifidobacteria is 7.0 × 109CFU/g。
(4) determination of incubation time
Inoculating proportion have been determined as three kinds of bacterium inoculate simultaneously viable count the highest under conditions of, by Lactobacillus plantarum
The glycerol pipe of TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 is inoculated in activation training by 1% respectively
Supporting base cultivation and once activate 18h, 37 DEG C, pH is natural, transfers into identical secondary by the bacterium amount that connects of 10%
In activation medium, carrying out re-activation 20h, 37 DEG C, pH is natural, in solid-state fermentation culture medium (by red
After bean, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts are cleaned, enter by material-water ratio 1:1 (mass volume ratio)
Row mixing, soaks 10h, 121 DEG C of sterilizing 20min at 4 DEG C) in inoculate three kinds of bacterial strains simultaneously and carry out mixed bacterium solid-state
Fermentation.The inoculum concentration of Lactobacillus plantarum TK9 and bacillus bifidus 937 is 1 × 107CFU/g, bacillus subtilis
Inoculum concentration be 5 × 104CFU/g, under 37 DEG C of .pH natural conditions of cultivation temperature, cultivate respectively 24h, 36h,
Respectively the culture medium after solid fermentation is carried out viable lactic acid bacteria counting after 48h, 60h.
The total viable count of lactic acid bacteria in table 6 solid fermentation food
Result is: probiotic bacteria five elements' bean optimal incubation time of solid fermentation food is 48h, records lactic acid bacteria and always lives
Bacterium number is 2.62 × 1010, wherein Lactobacillus plantarum TK9 viable count is 1.75 × 1010, animal bifidobacteria 937
Viable count is 8.70 × 109。
(5) active component change before and after fermentation
By the glycerol pipe of Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis TK-1 respectively
Being inoculated in activation medium by 1% to cultivate and once activate 18h, 37 DEG C, pH is natural, by 10% connect bacterium
Amount is transferred in identical re-activation culture medium, carries out re-activation 20h, and 37 DEG C, pH is natural, in solid-state
Fermentation medium is (after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, by material-water ratio 1:1.5
(mass volume ratio) mixes, at 4 DEG C soak 9h, 121 DEG C of sterilizing 20min) in inoculate three kinds of bacterium simultaneously
Strain carries out mixed fungus fermentation.
The inoculum concentration of Lactobacillus plantarum TK9 and bacillus bifidus 937 is 1 × 107CFU/g, bacillus subtilis
The inoculum concentration of TK-1 is 5 × 104CFU/g, cultivation temperature 37 DEG C, after cultivating 48h under pH natural conditions,
Measure the change of each active component content in solid fermentation thing.And by laggard for-70 DEG C of pre-freeze 6h of product by solid-state fermentation
Row vacuum lyophilization 24h, is not more than 5% to water content and obtains five elements' bean solid fermentation food.
Use same method, by Lactobacillus plantarum TK9, animal bifidobacteria 937 and bacillus subtilis
TK-1 replaces with Lactobacillus plantarum CGMCC 1.6971, animal bifidobacteria CGMCC 1.3003 and respectively
Bacillus subtilis CGMCC 1.3358, repeats above-mentioned experiment, result such as table 8.
GB/T 23788-2009 method is utilized to measure isoflavone content；GB/T 12143.2-89 method is utilized to survey
Determine the content of amino nitrogen；Utilize Syrups by HPLC GABA content；Utilize forint phenol reaction assay total
Phenols content；Oxygen index method (ORAC) of living is utilized to measure polyphenoils content.
The mensuration of active substance in table 7 solid fermentation food
Table 8
In summary it can be seen, the five elements after Lactobacillus plantarum, animal bifidobacteria and fermentation of bacillus subtilis
Viable count of lactobacillus in bean liquid beverage and solid-state food is higher than viable count measured during single bacterium fermentation, and
In tunning, active substance such as daidzein, genistein content dramatically increase, and this composition has prevention painstaking effort
Pipe disease, the effect of osteoporosis and prophylaxis of cancer improved；Total phenols and polyphenoils after mixed fungus fermentation contain
Amount is also obviously improved, this material can antioxidation, strengthening blood vessel wall, promote gastrointestinal disturbances, reduce blood fat and increasing
Add passive protective physical fitness, and the formation of arteriosclerosis and thrombosis can be prevented；And the γ-aminobutyric acid in tunning
(GABA) hypertension had alleviate and effect for the treatment of, it is seen that five elements' bean beverage after probiotics fermention and
Food has certain health-care effect and compares the fermented product of single beans, and five elements' bean fermented beverage and food can
Nourish the five internal organs simultaneously, will Semen Phaseoli blood pressure lowering reinforcing the heart, Semen phaseoli radiati removing toxic substances tonifying liver, Semen Glycines replenishing QI to invigorate the spleen, Semen Vignae Cylindricae help digestion profit
Effect of lung and Semen sojae atricolor enriching yin and nourishing kidney combines, nutrition more horn of plenty, functional the most extensive.Especially
It it is gained after the mixed fermentation of Lactobacillus plantarum 937, animal bifidobacteria TK9 and bacillus subtilis TK-1
In product, the content of active substance is more preferably.
Above-mentioned detailed with what food and preparation method thereof was carried out to this probiotics fermention five elements' bean beverage with reference to embodiment
Thin describe, be illustrative rather than determinate, can according to restriction scope list several embodiments,
Therefore changing and modifications under without departing from present general inventive concept, within should belonging to protection scope of the present invention.
Claims (12)
1. probiotics fermention five elements' bean food, it is characterised in that using five elements' bean as fermentation substrate, uses bacillus subtilis
At least two in bacterium, Lactobacillus plantarum and animal bifidobacteria is prepared after carrying out mixed bacterium solution state or solid fermentation.
2. a kind of probiotics fermention five elements' bean food as claimed in claim 1, it is characterised in that described Lactobacillus plantarum is for planting
Thing lactobacillus (Lactobacillus plantarum) TK9, deposit number is CGMCC No.11891；Described animal bifidobacteria is
Animal bifidobacteria (Bifidobacterium animalis) 937, deposit number is CGMCC No.11892；Described hay spore
Bacillus is bacillus subtilis (Bacillus subtilis) TK-1, and deposit number is CGMCCNo.4731.
3. a kind of probiotics fermention five elements' bean food as claimed in claim 1 or 2, it is characterised in that described probiotics fermention
The preparation method of five elements' bean food is as follows:
(1) actication of culture: bacillus subtilis, Lactobacillus plantarum and animal bifidobacteria are respectively from glycerol pipe, and pH is natural,
Cultivation temperature is 36-38 DEG C, incubation time 18-20h, and inoculum concentration 1% accesses activation medium and once activates, as one-level
Seed liquor；Cultivation accesses in identical activation medium by 10% inoculum concentration, in 36-38 DEG C, under pH natural conditions again after completing
Activation 20-24h, as secondary seed solution；
Described activation medium is: after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, in every 100mL water
Add the bean or pea making beating homogenizing after 5g mixed in equal amounts, homogenizing temperature 65 DEG C, pressure 20.0MPa；
(2) liquid fermentation is cultivated: bacillus subtilis, Lactobacillus plantarum and animal bifidobacteria are connect from secondary seed solution respectively
Plant and carry out any two bacterium or the mixed fermentation of three kinds of bacterium to liquid fermentation culture medium, 36-38 DEG C, under pH7 natural conditions, mix bacterium solution state
Standing for fermentation 12～24h；
(3) after acid adjustment, perfumery, homogenizing, sterilization, five elements' bean liquid fermentation beverage is obtained after fermentation ends.
4. a kind of probiotics fermention five elements' bean food, it is characterised in that described liquid fermentation culture medium is:
Pulverizing after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, after crossing 100 mesh sieves, by mass volume ratio 5-10%
Addition, mix homogeneously with water, 121 DEG C of sterilizing 20min.
5. a kind of probiotics fermention five elements' bean food, it is characterised in that described Lactobacillus plantarum and bifid
The inoculum concentration of bacillus is 5 × 107CFU/ml, the inoculum concentration of bacillus subtilis is 5 × 104CFU/ml。
6. as claimed in claim 3 a kind of probiotics fermention five elements' bean food, it is characterised in that the described mixed fungus fermentation time is
24h。
7. a kind of probiotics fermention five elements' bean food, it is characterised in that described five elements' bean liquid fermentation is drunk
The water content of material is not more than 95%.
8. a kind of probiotics fermention five elements' bean food as claimed in claim 1 or 2, it is characterised in that described probiotics fermention
The preparation method of five elements' bean food is as follows:
(1) actication of culture: bacillus subtilis, Lactobacillus plantarum and animal bifidobacteria are respectively from glycerol pipe, inoculum concentration
1% accesses activation medium once activates, and pH is natural, and cultivation temperature is 36-38 DEG C, incubation time 18-20h, as
Primary seed solution；Cultivation accesses in identical activation medium by 10% inoculum concentration, in 36-38 DEG C, under pH natural conditions after completing
Again activate 20-24h, as secondary seed solution；
Described activation medium is: after Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts being cleaned, in every 100mL water
Add the bean or pea making beating homogenizing after 5g mixed in equal amounts, homogenizing temperature 65 DEG C, pressure 20.0MPa；
(2) solid state fermentation culture: the secondary seed solution in step (1) is accessed in solid-state fermentation culture medium carry out any two bacterium and
The mixed fermentation of three kinds of bacterium, at 36-38 DEG C, mixed culture solid state fermentation 24-48h obtains five elements' bean probiotic bacteria solid fermentation food.
9. a kind of probiotics fermention five elements' bean food, it is characterised in that described solid-state fermentation culture medium is:
After Semen Phaseoli, Semen phaseoli radiati, Semen sojae atricolor, Semen Vignae Cylindricae, Semen Glycines mixed in equal amounts are cleaned, mix by material-water ratio 1:1-1.5, soak at 4 DEG C
8-10h prepares, 121 DEG C of sterilizing 20min.
10. a kind of probiotics fermention five elements' bean food, it is characterised in that plant in described solid fermentation
The inoculum concentration of lactobacillus and bacillus bifidus is 1 × 107CFU/g, the inoculum concentration of bacillus subtilis is 5 × 104CFU/g。
11. a kind of probiotics fermention five elements' bean food, it is characterised in that described mixed fungus fermentation time 48h.
12. a kind of probiotics fermention five elements' bean food, it is characterised in that after described solid fermentation terminates
Carrying out vacuum lyophilization, condition is-70 DEG C of pre-freeze 6h final vacuum lyophilization 24h, is not more than 5% to water content, obtains the five elements
Bean probiotic bacteria solid fermentation food.
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