CN108504609A - A kind of method of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentation - Google Patents
A kind of method of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentation Download PDFInfo
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- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
Abstract
A kind of method of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentation, picking bacillus coagulans inclined-plane seed and bacillus coagulans inclined-plane seed in seed fluid nutrient mediums of saccharomycete respectively at being cultivated, solid fermentation culture medium after sterilizing, by 5% 10% inoculation bacillus coagulans seed liquors of solid fermentation culture medium weight, by 3% 5% inoculation saccharomyces cerevisiae seed liquors of solid fermentation culture medium weight, the material after bacterium will be connect to be placed in the semiclosed device of taper ampuliform made of gas impermeable material, 35 37 DEG C of cultivation temperature, 44 48h of incubation time；Fermentation ends, 55 DEG C of drying, crush, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.Advantage is：In mixed culture solid state fermentation, two plants of bacterium mutualisms, bacterium number, enzyme activity, production acid and the substrate utilization ratio aspect of mixed fermentation are significantly higher than the solid state fermentation of single strain, have the advantages that equipment investment is few, energy saving, technique is simplified, easy to operate.
Description
Technical field
The invention belongs to technical field of microbial fermentation, it is related to a kind of bacillus coagulans and saccharomyces cerevisiae mixes bacterium solid-state hair
The method of ferment.
Background technology
Saccharomyces cerevisiae（saccharomyces cerevisiae）It is the Ministry of Agriculture of China to allow to make in No. 318 bulletins for 2003
One of microorganism fungus kind is a kind of single celled eukaryotic microorganism, played a role by alimentary canal in the form of viable bacteria, can be notable
The work for improving the amylase, lipase and pepsin of animal intestinal tract improves feed report to improve the digestion power of animal
Reward has stronger immunoloregulation function, reduces cholesterol and antioxidation, and saccharomycete can be resistant to the anti-of doses
Raw element, can gradually help feed transition of the animal from antibiotic to antibiotic-free.
Bacillus coagulans（bacillus coagulans）, the new edition of the Ministry of Agriculture of China in December, 2013 announcement《Feed
Additive kind catalogue（2013）》It is formally included in, pig, chicken, aquatic livestock are used in, in addition to resistance to peracid, high heat resistance,
The characteristics of bacillus such as osmophilic strain, easy to maintain, lactic acid is also generated, it is dual excellent with lactic acid bacteria and bacillus
Characteristic, shelf-stable, non-easy in inactivation can promote growth of animal, improve animal product quality, adjust intestinal flora balance, stimulation
Immune etc. significant effect, is one of dominant bacteria of substitute antibiotics.
Currently, have been reported the mixed culture solid state fermentation method of bacillus coagulans, mixed strain is with lactobacillus plantarum, dry
Based on Lactobacillus paracasei and bacillus subtilis, and most is multi-cultur es step fermentation.Bacillus coagulans and saccharomyces cerevisiae mix bacterium
Process for solid state fermentation rare report.
Invention content
The technical problem to be solved in the present invention is to provide a kind of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentations
Method, in mixed culture solid state fermentation, two plants of bacterium mutualisms, bacterium number, enzyme activity, production acid and the substrate utilization ratio side of mixed fermentation
Face is significantly higher than the solid state fermentation of single strain, has the advantages that equipment investment is few, energy saving, technique is simplified, easy to operate.
Technical solution of the invention is：
A kind of method of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentation, comprises the concrete steps that：
（1）The culture of bacillus coagulans strain
Seed one ring in picking bacillus coagulans inclined-plane is cultivated in seed fluid nutrient mediums of saccharomycete, 35 DEG C -40 DEG C of cultivation temperature,
Rotating speed 160-200rpm, incubation time 18-24h；
（2）The culture of saccharomyces cerevisiae
Seed one ring in picking bacillus coagulans inclined-plane is cultivated in seed fluid nutrient mediums of saccharomycete, 30 DEG C -35 DEG C of cultivation temperature,
Rotating speed 160 rpm -200rpm, incubation time 16h-24h；
（3）Make solid fermentation culture medium
Solid-state fermentation culture medium weight ratio group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:(6-10): (2-4):(3-
4), moisture control is in 40%-65%, pH 6.8-7.2；
（4）Solid fermentation culture
Solid fermentation culture medium after sterilizing is inoculated with bacillus coagulans seed by the 5%-10% of solid fermentation culture medium weight
Liquid is inoculated with saccharomyces cerevisiae seed liquor by the 3%-5% of solid fermentation culture medium weight, will connect the material after bacterium and is placed in gas impermeable material
In the semiclosed device of manufactured taper ampuliform, 35-37 DEG C of cultivation temperature, incubation time 44-48h.Fermentation ends, 55 DEG C of bakings
It is dry, it crushes, sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.
Further, the culture medium of the seed of the bacillus coagulans and saccharomyces cerevisiae, g/L：Glucose 20-50, egg
White peptone 10-50, yeast extract 10-50, pH 6.8-7.2.
Further, when solid fermentation culture, culture apparatus contains the weight of culture medium and the volume ratio of culture apparatus is
1:5 g/mL -1:30 g/mL。
Further, the bacillus coagulans are bacillus coagulans LCCC10025 purchased in market, and saccharomyces cerevisiae is purchased in market
Saccharomyces cerevisiae LCCC20003, bacillus coagulans LCCC10025 are preserved in Liaoning Province's Culture Collection, and number is
LCCC10025, saccharomyces cerevisiae LCCC20003 are preserved in Liaoning Province's Culture Collection, number LCCC20003.
Further, Bacillus coagulans spore number >=2.0 in the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae
×109CFU/g, yeast count >=1.4 × 109CFU/g。
Beneficial effects of the present invention：
1. two plants of bacterium mutualisms in mixed fermentation, saccharomyces cerevisiae can provide anaerobic environment for the growth of bacillus coagulans, coagulate
Knot bacillus provides nutriment for the growth of saccharomyces cerevisiae, and Hybrid NC machine tool method can obtain higher bacillus coagulans
With the viable count of saccharomyces cerevisiae.Bacillus coagulans spore number >=2.0 × 10 in the composite bacteria agent that the fermentation process obtains9CFU/
G, yeast count >=1.4 × 109CFU/g。
2, semiclosed training method meets the demand of bacillus coagulans and saccharomyces cerevisiae to oxygen simultaneously.This method
Rice husk ingredient is introduced in solid medium, to adjust the ventilatory capacity needed for growth.
3, bacillus coagulans and saccharomyces cerevisiae mixed fermentation secrete higher protease and amylase in growth course,
The composite bacteria agent prepared by this method is remarkably improved the digestion power and the price of deed of animal；The composite bacteria agent of production was both
There is the ester fragrance of yeast, and have the tart flavour of bacillus coagulans, palatability is good.
4, using one-step method solid-state mixed fermentation bacillus coagulans and saccharomyces cerevisiae, simple, small investment, work with equipment
Skill simple and convenient operation flow, the advantage that low energy consumption.
Specific implementation mode
With reference to example, the present invention is further described, and following examples are illustrative, are not restrictive, cannot
Protection scope of the present invention is limited with following examples.
1, prepared by the seed liquor of strain
The culture medium of the seed of the bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 20-50, peptone 10-50, ferment
Female cream 10-50, pH 6.8-7.2；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；It is purchased in market, bacillus coagulans
LCCC10025 is in Liaoning Province's Culture Collection, and deposit number LCCC10025, saccharomyces cerevisiae LCCC20003 is in the Liao Dynasty
It is peaceful to save Culture Collection, deposit number LCCC20003；
The condition of culture of the strain is：
Bacillus coagulans：35-40 DEG C of cultivation temperature, rotating speed 160-200rpm, incubation time 18-24h；
Saccharomyces cerevisiae：30-35 DEG C of cultivation temperature, rotating speed 160-200rpm, incubation time 16-24h；
2, mixed culture solid state fermentation
The solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:(6-10): (2-
4):(3-4), moisture are controlled in 40%-60%, pH 6.8-7.2.
The solid state fermentation cultural method is：Culture medium after sterilizing is inoculated with bacillus coagulans by the 5%-10% of its weight
Seed liquor is inoculated with saccharomyces cerevisiae seed liquor by the 3%-5% of its weight, will connect the material after bacterium and is placed in made of gas impermeable material and bores
In the semiclosed device of shape ampuliform, 35 DEG C -37 DEG C of cultivation temperature, incubation time 44h-48h.Fermentation ends, 55 DEG C of drying, powder
Broken, sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.
The assay method of viable count of the embodiment of the present invention
Counting method is coated with using dilution plate, operating procedure is：The composite bacteria agent 10g prepared is weighed, the sterile of 90ml is placed in
In physiological saline triangular flask, in stirring 20min on magnetic stirring apparatus, the bacterium solution of 1ml is taken to be added to 9ml sterile physiologicals with liquid-transfering gun
In the test tube of brine, ten times of gradient dilutions are taken turns doing.It takes the dilution 0.1ml of suitable gradient in the culture medium solidified, applies
Cloth is uniform, does three Duplicate Samples, is placed in 37 DEG C of incubators and cultivates 48h.
Embodiment 1
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10,
pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：38 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:4:4, moisture
60%, pH 7；
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with by 5% inoculation bacillus coagulans seed liquor of culture medium weight by the 5% of culture medium weight
Saccharomyces cerevisiae seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, cultivates
The weight of base and the volume ratio of culture apparatus are 1:15（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55
DEG C drying, crush, sieving, obtain the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.Bacillus coagulans spore number 3.5
×109CFU/g, yeast count 6.2 × 109CFU/g。
Embodiment 2
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 20, peptone 10, yeast extract 12,
pH 6.8；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：35-40 DEG C of cultivation temperature, rotating speed 160-200rpm, incubation time 18-24h；
Saccharomyces cerevisiae：30 DEG C, rotating speed 160rpm, incubation time 16h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:4:4, moisture
60%, pH 7.
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 7% inoculation bacillus coagulans seed liquor of its weight by the 3% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:15（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.Bacillus coagulans spore number 9.5 ×
109CFU/g, yeast count 1.4 × 109CFU/g。
Embodiment 3
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 50, peptone 50, yeast extract 50,
pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：35 DEG C, rotating speed 200rpm, incubation time 18h of cultivation temperature；
Saccharomyces cerevisiae：35 DEG C of cultivation temperature, rotating speed 200rpm, incubation time is for 24 hours；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:4:4, moisture
60%, pH 7.2.
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 6% inoculation bacillus coagulans seed liquor of its weight by the 4% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:15（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h.Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.Bacillus coagulans spore number 6.5 ×
109CFU/g, yeast count 2.8 × 109CFU/g。
4 bacillus coagulans of embodiment and saccharomyces cerevisiae mixed solid fermentation method（Culture medium different proportion formula）
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10,
pH 7.2；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：38 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:9:2:3, moisture
60%, pH 7；
Solid state fermentation culture：Culture medium after sterilizing is inoculated with bacillus coagulans seed liquor by the 7% of its weight, by its weight
3% inoculation saccharomyces cerevisiae seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material
In, the weight of the culture medium filled and the volume ratio of culture apparatus are 1:5（g/mL）, 35 DEG C of cultivation temperature, incubation time
44h；Fermentation ends, 55 DEG C of drying, crush, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae；Compound bacteria
Agent kind Bacillus coagulans spore number 7.5 × 109CFU/g, yeast count 3.2 × 109CFU/g。
5 bacillus coagulans of embodiment and saccharomyces cerevisiae mixed solid fermentation method（Culture medium different proportion formula）
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10,
pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：38 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:6:3:4, moisture
60%, pH 6.8；
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 6% inoculation bacillus coagulans seed liquor of its weight by the 4% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:30（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae；Composite bacteria agent kind bacillus coagulans
Gemma number 12.6 × 109CFU/g, yeast count 2.9 × 109CFU/g。
6 bacillus coagulans of embodiment and saccharomyces cerevisiae mixed solid fermentation method（Culture medium different proportion formula）
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10,
pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：38 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:4:4, moisture
50%, pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 7% inoculation bacillus coagulans seed liquor of its weight by the 3% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:10（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h.Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.Composite bacteria agent kind bacillus coagulans
Gemma number 6.8 × 109CFU/g, yeast count 4.6 × 109CFU/g。
7 bacillus coagulans of embodiment and saccharomyces cerevisiae mixed solid fermentation method（Culture medium different proportion formula）
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10,
pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：
Bacillus coagulans：38 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:4:4, moisture
65%, pH 7；
Solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 7% inoculation bacillus coagulans seed liquor of its weight by the 3% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:10（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae；Composite bacteria agent kind bacillus coagulans
Gemma number 9.9 × 109CFU/g, yeast count 3.8 × 109CFU/g。
The solid state rheology of 1 single bacillus coagulans of comparative example
（1）The culture medium of bacillus coagulans seed（g/L）：Glucose 25, peptone 15, yeast extract 10, pH7；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：Bacillus coagulans：38 DEG C of cultivation temperature, rotating speed
180rpm, incubation time 20h；
（2）Solid-state fermentation culture medium ratio group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:1:1, moisture 50%, pH
7；
Strain selects：Bacillus coagulans LCCC10025；
Solid state fermentation culture：
Culture medium after sterilizing by its weight 5% inoculation bacillus coagulans seed liquor, will connect the material after bacterium be placed in it is airtight
In the semiclosed device of taper ampuliform made of material, the weight of the culture medium filled and the volume ratio of culture apparatus are 1:15
（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h.Fermentation ends, 55 DEG C of drying, crush, and sieving obtains bacillus coagulans
Microbial inoculum, Bacillus coagulans spore, 3.1 × 108CFU/g。
The solid state rheology of 2 single saccharomyces cerevisiae of comparative example.
（1）The culture medium of the seed of saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract 10, pH 7；
Strain selects：Saccharomyces cerevisiae LCCC20003；
The seed culture condition of saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 20h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:10:1:1, moisture 50%, pH
7；
Solid state fermentation culture：
Culture medium after sterilizing will be connect the material after bacterium and be placed in gas impermeable material by 5% inoculation saccharomyces cerevisiae seed liquor of its weight
In the semiclosed device of manufactured taper ampuliform, the weight of the culture medium filled and the volume ratio of culture apparatus are 1:10（g/
mL）, 30 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55 DEG C of drying, crush, and sieving obtains saccharomyces cerevisiae microbial inoculum.Ferment
Female bacterium number 8.7 × 108CFU/g。
3 bacillus coagulans of comparative example and saccharomyces cerevisiae mixed solid fermentation method（Rice husk is not added in culture medium prescription）.
（1）The culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae（g/L）：Glucose 25, peptone 15, yeast extract
10,pH 7；
Strain selects：Bacillus coagulans LCCC10025 and saccharomyces cerevisiae LCCC20003；
The seed culture condition of bacillus coagulans and saccharomyces cerevisiae：Bacillus coagulans：38 DEG C of cultivation temperature, rotating speed
180rpm, incubation time 20h；
Saccharomyces cerevisiae：32 DEG C, rotating speed 180rpm, incubation time 18h of cultivation temperature；
（2）Solid-state fermentation culture medium ratio（Weight）Group becomes：Maize flour：Wheat bran：Beancake powder=10:10:1, moisture 60%, pH
7。
The solid state fermentation culture：
Culture medium after sterilizing is inoculated with saccharomyces cerevisiae by 5% inoculation bacillus coagulans seed liquor of its weight by the 5% of its weight
Seed liquor will connect the material after bacterium and be placed in the semiclosed device of taper ampuliform made of gas impermeable material, the culture medium filled
Weight and culture apparatus volume ratio be 1:20（g/mL）, 35 DEG C of cultivation temperature, incubation time 44h；Fermentation ends, 55 DEG C
Drying crushes, and sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae；Bacillus coagulans spore number 7.5 ×
108CFU/g, yeast count 9.8 × 108CFU/g。
It can show that Hybrid NC machine tool method can obtain by 1- of embodiment of the present invention embodiments 7 and comparative example 1- comparative examples 2
The viable count of higher bacillus coagulans and saccharomyces cerevisiae, bacillus coagulans bud in the composite bacteria agent which obtains
Spore number >=2.0 × 109CFU/g, yeast count >=1.4 × 109CFU/g.Two plants of bacterium mutualisms, saccharomyces cerevisiae in mixed fermentation
Anaerobic environment can be provided for the growth of bacillus coagulans, bacillus coagulans provide nutriment for the growth of saccharomyces cerevisiae,
Thus, it is possible to obtain higher viable count.
It can be obtained by 1- of embodiment of the present invention embodiments 7 and comparative example 3, the addition of rice husk in solid medium, it can be with
The viable count of higher bacillus coagulans and saccharomyces cerevisiae is obtained, rice husk ingredient is used in solid medium, can adjust solidifying
Bacillus and saccharomyces cerevisiae growth are tied to the demand of oxygen, the ventilatory capacity needed for growth is adjusted, meets condensation gemma
The demand of bacillus and saccharomyces cerevisiae to oxygen.
Claims (5)
1. a kind of method of bacillus coagulans and saccharomyces cerevisiae mixed culture solid state fermentation, it is characterized in that：
It comprises the concrete steps that：
（1）The culture of bacillus coagulans strain
Seed one ring in picking bacillus coagulans inclined-plane is cultivated in seed fluid nutrient mediums of saccharomycete, 35 DEG C -40 DEG C of cultivation temperature,
Rotating speed 160-200rpm, incubation time 18-24h；
（2）The culture of saccharomyces cerevisiae
Seed one ring in picking bacillus coagulans inclined-plane is cultivated in seed fluid nutrient mediums of saccharomycete, 30 DEG C -35 DEG C of cultivation temperature,
Rotating speed 160 rpm -200rpm, incubation time 16h-24h；
（3）Make solid fermentation culture medium
Solid-state fermentation culture medium weight ratio group becomes：Maize flour：Wheat bran：Beancake powder：Rice husk=10:(6-10): (2-4):(3-
4), moisture control is in 40%-65%, pH 6.8-7.2；
（4）Solid fermentation culture
Solid fermentation culture medium after sterilizing is inoculated with bacillus coagulans seed by the 5%-10% of solid fermentation culture medium weight
Liquid is inoculated with saccharomyces cerevisiae seed liquor by the 3%-5% of solid fermentation culture medium weight, will connect the material after bacterium and is placed in gas impermeable material
In the semiclosed device of manufactured taper ampuliform, 35-37 DEG C of cultivation temperature, incubation time 44-48h；Fermentation ends, 55 DEG C of bakings
It is dry, it crushes, sieving obtains the composite bacteria agent containing bacillus coagulans and saccharomyces cerevisiae.
2. the method for bacillus coagulans according to claim 1 and saccharomyces cerevisiae mixed culture solid state fermentation, it is characterized in that：Institute
State the culture medium of the seed of bacillus coagulans and saccharomyces cerevisiae, g/L：Glucose 20-50, peptone 10-50, yeast extract 10-
50, pH 6.8-7.2.
3. the method for bacillus coagulans according to claim 1 and saccharomyces cerevisiae mixed culture solid state fermentation, it is characterized in that：Gu
When body fermented and cultured, it is 1 that culture apparatus, which contains the weight of culture medium and the volume ratio of culture apparatus,:5 g/mL -1:30 g/mL.
4. the method for bacillus coagulans according to claim 1 and saccharomyces cerevisiae mixed culture solid state fermentation, it is characterized in that：Institute
The bacillus coagulans stated are bacillus coagulans LCCC10025, and saccharomyces cerevisiae is saccharomyces cerevisiae LCCC20003.
5. the method for bacillus coagulans according to claim 1 and saccharomyces cerevisiae mixed culture solid state fermentation, it is characterized in that：Contain
Bacillus coagulans spore number >=2.0 × 10 in the composite bacteria agent of bacillus coagulans and saccharomyces cerevisiae9CFU/g, yeast count
≥1.4×109CFU/g。
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| CN112175835A (en) * | 2019-07-05 | 2021-01-05 | 中粮生物化学（安徽）股份有限公司 | Application of lactobacillus plantarum in storage of bacillus subtilis fermentation liquor |
| CN113826757A (en) * | 2021-08-12 | 2021-12-24 | 陕西杨凌富仕特生物科技有限公司 | Yeast culture rich in active probiotics, preparation method and special equipment thereof |
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| CN113826757B (en) * | 2021-08-12 | 2024-03-26 | 陕西杨凌富仕特生物科技有限公司 | Yeast culture rich in active probiotics, preparation method and special equipment thereof |
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