CN108893433A - A kind of crop material waste leavening and its preparation method and application - Google Patents
A kind of crop material waste leavening and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a kind of crop material waste leavenings and its preparation method and application, belong to agricultural technology field, the leavening is made of Brevibacillus laterosporus bacterium powder, S. cervisiae powder, thermophilic sporotrichum bacterium powder and Geotrichum powder, and the bacteria concentration of each bacterium is 10 in the leavening7‑109Cfu/g；The Geotrichum powder is made by following methods：It after geotrichum candidum conidia powder and yam flour are mixed, is inoculated on solid medium, the solid medium after inoculation is then subjected to koji plate fermentation, after the koji plate fermentation, obtain Geotrichum powder through drying and crushing.The leavening has the characteristics that composite construction property is stable, antagonism is small, enzymatic productivity is strong, functional flora effect is with specific directionality, the effect of it can give full play to the effect of the synergistic combinations between the unique effect of function bacterium and flora, be finally reached quickly thoroughly fermentation crop material waste.It is used for agricultural, the economic benefit and environmental benefit having had both.
Description
Technical field
The invention belongs to agricultural technology fields, and in particular to a kind of crop material waste leavening and preparation method thereof and
Using.
Background technique
Stalk is a kind of Biomass Energy Resources, is the fourth-largest energy that coal, oil and natural gas are only second in the world today
Source, and unique reproducible energy.But China's stalk resource is burnt up mostly with fuel forms, only a small part passes through
Cross abdomen returning to the field or direct returning to farmland.And stalk directly burns and not only pollutes environment, but also will cause the wasting of resources.
The directly edible stalk of animal is the excellent approach of fodder, but stalk is directly edible, can there is such as straw utilization rate
It is low, it is nondigestible, influence the problems such as growing.And utilize venomous injurant most of in microbial fermentation then degradable stalk
Matter will be not easy the ammonia that the complicated macromolecules degradation such as protein, cellulose, starch for being digested and assimilated by animal is absorption easy to digest
The small molecules such as base acid, polypeptide, organic acid, oligosaccharides, and the mycoprotein for being easy to be utilized is formed, to greatly improve the battalion of stalk
Support value and utilization efficiency.Meanwhile the palatability of feed can be improved by the aromatic substance that fermentation generates, promote feed intake,
The bioactive substances such as the beneficial microbe being largely proliferated and its produced organic acid, enzyme, unknown growth factor can promote animal
The growth of internal beneficial microbe seedling group and the digestibility and utilization of nutriment, inhibit harmful bacteria, to safeguard animal health, reduce
Epidemic disease occurs and drug uses
But currently, equally existing various particular problems and deficiency using microbial fermentation, such as technology is immature, efficiency is not
Height, strain are excellent etc..The technical principle of existing crop material waste leavening only according in crop material waste such as
Amylase, protease, cellulase, hemicellulose are produced in the ingredients such as starch, albumen, cellulose, hemicellulose, lignin, screening
The microbial strains of plain enzyme, lignoenzyme etc. carry out microorganism compounding, them are made effectively to be combined with each other.However, existing institute
In the leavening product of production, most of leavening products are deposited both for the microbial inoculum product of development & production under the conditions of banking up
In the disadvantage that product bacterium number content is low, enzyme activity quantity is too small, it is difficult to meet the actual needs of market application, while leavening produces
The strain and strain that product are selected assemble unreasonable, could not fully consider the phase between the enzymatic productivity and strain of strain itself
Mutually there is the influence of symbiosis, alternate, antagonism to product, especially fungal species are few, crop material waste in use
Ferment effect stability is poor.
Therefore, it in order to provide stalk digestibility, develops a circular economy, realizes sustainable development, it is necessary to develop a kind of efficient
Excellent microbial bacterial agent.
Summary of the invention
In view of this, one of the objects of the present invention is to provide a kind of crop material waste leavenings；The second purpose exists
In providing a kind of preparation method of crop material waste leavening；The third purpose is to provide crop material waste leavening
Preparing the application in biological feedstuff.
In order to achieve the above objectives, the present invention provides the following technical solutions：
1, a kind of crop material waste leavening, the leavening is by Brevibacillus laterosporus bacterium powder, S. cervisiae
Powder, thermophilic sporotrichum bacterium powder and Geotrichum powder form, and the bacteria concentration of each bacterium is 10 in the leavening7-109Cfu/g；Institute
Geotrichum powder is stated to be made by following methods：After geotrichum candidum conidia powder and yam flour are mixed, it is inoculated on solid medium, so
The solid medium after inoculation is subjected to koji plate fermentation afterwards, after the koji plate fermentation, obtains Geotrichum through drying and crushing
Powder.
Preferably, the weight ratio of the geotrichum candidum conidia powder, yam flour and solid medium is 0.01-0.02:1-2:1-
2。
Preferably, the granular size of the yam flour is 60-200 mesh.
Preferably, the solid medium is prepared by following methods：It is in mass ratio by wheat bran, bean cake powder and corn flour
50-70:15-20:Then the epsom salt, 2-3.5% for being equivalent to mixture weight 0.3-0.5% is added in 15-25 mixing
The potassium dihydrogen phosphate of ferric sulfate and 0.1-0.2%, finally adding water to biodiversity percentage is 50-60%.
2, the preparation method of a kind of crop material waste leavening, described method includes following steps：
(1) Brevibacillus laterosporus is seeded in the fermentor containing culture medium and carries out fermented and cultured, the fermentation training
After supporting, fermentation liquid is concentrated first, obtains concentrated broth, lightweight carbon is then added into the concentrated broth
Sour calcium is spray-dried after stirring evenly, and obtains Brevibacillus laterosporus bacterium powder；
(2) saccharomyces cerevisiae is seeded in the fermentor containing culture medium and carries out fermented and cultured, the fermented and cultured terminates
Afterwards, fermentation liquid is concentrated first, obtains concentrated broth, precipitated calcium carbonate then is added into the concentrated broth,
It is spray-dried after stirring evenly, obtains S. cervisiae powder；
(3) it after mixing thermophilic sporotrichum conidia powder and flour, is inoculated on solid medium, it then will be after inoculation
Solid medium carries out koji plate fermentation, after the koji plate fermentation, obtains thermophilic sporotrichum bacterium powder through drying and crushing；
(4) it after mixing geotrichum candidum conidia powder and yam flour, is inoculated on solid medium, then by the solid after inoculation
Culture medium carries out koji plate fermentation, after the koji plate fermentation, obtains Geotrichum powder through drying and crushing；
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
Preferably, in step (1), Brevibacillus laterosporus is seeded to the hair containing culture medium by the inoculum concentration of 1-5%
In fermentation tank.
Preferably, in step (1), the culture medium by mass percentage, is grouped as by following group：Glucose 0.01-
0.1%, beef extract 0.01-0.1%, peptone 0.01-0.1%, sodium chloride 0.01-0.1%, surplus are water, pH=6-7.5.
Preferably, in step (1), the culture medium by mass percentage, is grouped as by following group：Glucose
0.05%, beef extract 0.05%, peptone 0.05%, sodium chloride 0.05%, surplus is water, pH=6-7.5.
Preferably, in step (1), the fermented and cultured is passed through filtrated air specifically, under the conditions of 30-37 DEG C, culture
After 8 hours, start to sample microscopy, incubation time is taken a sample to check secondary, incubation time is 24 for every 4 hours within the scope of 8 to 24 hours
To within the scope of 36 hours, check within every 2 hours primary, it is ensured that without miscellaneous bacteria, when Brevibacillus laterosporus spore forming rate reaches 90-
When 95%, fermented and cultured is completed.
Preferably, in step (2), saccharomyces cerevisiae is seeded in the fermentor containing culture medium by the inoculum concentration of 1-5%.
Preferably, in step (2), the culture medium by mass percentage, is grouped as by following group：Glucose 0.01-
1.0%, yeast extract 0.01-1.0%, peptone 0.01-1.0%, bean cake powder 0.01-1.0%, potassium dihydrogen phosphate 0.01-
1.0%, surplus is water, pH=6-7.5.
Preferably, in step (2), the culture medium by mass percentage, is grouped as by following group：Glucose 0.5%,
Yeast extract 0.2%, peptone 0.1%, bean cake powder 0.5%, potassium dihydrogen phosphate 0.05%, surplus are water, pH=6-7.5.
Preferably, in step (2), the fermented and cultured is passed through filtrated air specifically, under the conditions of 30-37 DEG C, culture
After 8 hours, start to sample microscopy, incubation time is taken a sample to check secondary, incubation time is 24 for every 4 hours within the scope of 8 to 24 hours
It within the scope of 48 hours, checks within every 2 hours primary, it is ensured that without miscellaneous bacteria, after saccharomyces cerevisiae, which sprouts, to be stablized, fermented and cultured is completed.
Preferably, in step (1) and step (2), the concentration is specially 5-10 times of concentration.
Preferably, in step (1) and step (2), the additional amount of the precipitated calcium carbonate is the concentrated broth weight
20-40%.
Preferably, in step (1) and step (2), when the spray drying, intake air temperature is 195-200 DEG C, air outlet
Temperature is 75-80 DEG C, feed rate 5-20mL/min.
Preferably, in step (3), the weight ratio of the thermophilic sporotrichum conidia powder, flour and solid medium is 1:
10-50:10-50。
Preferably, in step (3) and step (4), the solid medium is prepared by following methods：By wheat bran, bean cake powder
It is in mass ratio 50-70 with corn flour:15-20:15-25 mixing, is then added and is equivalent to the seven of mixture weight 0.3-0.5%
The potassium dihydrogen phosphate of water magnesium sulfate, the ferric sulfate of 2-3.5% and 0.1-0.2%, finally adding water to biodiversity percentage is 50-
60%.
Preferably, in step (3) and step (4), the koji plate fermentation is specifically, the solid medium after inoculation is put into
It is that 6-10cm carries out paving disk by laying depth on the sterile indoor koji tray of fermentation, under conditions of ventilation, consolidating after keeping inoculation
The temperature of body culture medium is 30 DEG C, is fermented 35-50 hours.
3, a kind of crop material waste leavening is preparing the application in biological feedstuff.
The beneficial effects of the present invention are：The present invention provides a kind of crop material waste leavenings and preparation method thereof
And application, the leavening is by Brevibacillus laterosporus bacterium powder, S. cervisiae powder, thermophilic sporotrichum bacterium powder and Geotrichum powder
Composition, with composite construction property is stable, antagonism is small, enzymatic productivity is strong, functional flora effect has specific directionality
The features such as, which can give full play to the effect of the synergistic combinations between the unique effect of function bacterium and flora, be finally reached fast
The effect of fast thoroughly fermentation crop material waste.Leavening in the present invention is used for the fermentation of crop material waste, it can be with
Make time advance 49-54 days of stalk complete fermentation maturation, and crop material waste is after ferment-fermented in the present invention,
The feed intake and digestibility of the animals such as ox, sheep can be dramatically increased.In addition, due to N, P rich in crop material waste,
The inorganic nutrients such as K and organic matter abundant, by its through the invention in it is ferment-fermented after obtain biological feedstuff, the biological feedstuff
In contain a large amount of nutriment and probiotics, nutrients ratio more coordinates, and raises animal with the biological feedstuff, is not only able to
The usage amount of other feeds is reduced, animal product yield and quality can also be improved, and then increase farmers' income, use the present invention
The leavening economic benefit that can reach abridged edition, lower consumption, increase income.Finally, crop material waste through the invention in hair
Ferment agent is also avoided that in line caused environmental pollution while the biological feedstuff being converted to, and the leavening in the present invention can make
Agriculture " three wastes " are preferably recycled, and then play the role of improving ecological protection environment, are economized on resources, are effectively promoted
Into the sustainable development of agricultural.
In the leavening in the preparation process of Geotrichum powder, it is trained for the one of its nutrient media components with yam flour
It supports, can effectively enhance the fermentation and capacity of decomposition, the adaptability in animal and bird intestines and stability genetic breeding ability of the bacterium,
Wherein, the substances such as Chinese yam institute cellulase, starch multienzyme complex can be improved geotrichum candidum to stalk fibre during the fermentation
Fermentability, so that it is converted into starch, then under the action of amylase, be converted into glucose for animal use；In Chinese yam
Protein, starch, amylase, maltose etc. can be improved the flourish ability of geotrichum candidum and keeps its biology in crop material
Active ability；The substances such as choline contained by Chinese yam, mucus juice enzyme and Dioscin can prevent the non-of bacterium in geotrichum candidum fermentation
Hereditary variation makes it have stability genetic breeding ability, and can reduce the toxic side effect of geotrichum candidum metabolite.Phase
It answers, Chinese yam is after geotrichum candidum ferments, protein, carbohydrate, mucus juice, tannin, amino acid, polysaccharide in Chinese yam etc.
Substance can be converted into the acitve organic matter absorbed conducive to livestock and poultry, wherein after the fermentation of materials such as mucus juice, tannin, can make mountain
The adsorption capacity and increased activity of effective component in medicine, and the acidic materials that geotrichum candidum generates during the fermentation can change
The acid-base balance of kind crop material, can keep higher activity during the fermentation, play promotion fermentation, rapidly and efficiently
The effect of decomposition of cellulose.
Specific embodiment
Below by a preferred embodiment of the present invention will be described in detail.
Brevibacillus laterosporus (Brevibacillus laterosporu, ATCC in strain used in the present invention
10249), it is purchased from American Type Culture Collecti；Saccharomyces cerevisiae (Saccharomyces cerevisiae, ATCC 26603), purchase
In American Type Culture Collecti；Thermophilic sporotrichum (Sporotrichum thermophile, ACCC 30346), is purchased from China
Agriculture Organism Depositary；Geotrichum candidum (Cryytococcus neoformans, ATCC 10663) is purchased from U.S.'s strain guarantor
Hiding center.
Embodiment 1
Prepare a kind of crop material waste leavening
(1) Brevibacillus laterosporus bacterium powder is prepared
The Brevibacillus laterosporus strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature is controlled 30
DEG C, culture is transferred in the eggplant type bottle with culture medium again after 40 hours and cultivates 32 hours, and culture medium presses quality percentage in eggplant type bottle
Than meter, it is grouped as by following group：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.2.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 32 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.05%, beef extract 0.05%, peptone 0.05%, sodium chloride 0.05%,
Surplus is water, pH=7.2.
Brevibacillus laterosporus is seeded in the fermentor containing culture medium by 5% inoculum concentration, under the conditions of 32 DEG C,
It is passed through filtrated air, after culture 8 hours, starts to sample microscopy, incubation time is within the scope of 8 to 24 hours, the inspection of sampling in every 4 hours
It looks into time, incubation time checks primary, it is ensured that within the scope of 24 to 36 hours without miscellaneous bacteria, when Brevibacillus laterosporus bud for every 2 hours
When spore formation rate reaches 95%, fermented and cultured is completed.
Fermentation liquid is subjected to 8 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 32% is added in liquid, in intake air temperature is 196 DEG C after stirring evenly, air outlet temperature is
80 DEG C, feed rate is spray-dried under conditions of being 12mL/min, obtains Brevibacillus laterosporus bacterium powder, bacterium in the bacterium powder
Content is 109Cfu/g。
(1) S. cervisiae powder is prepared
The saccharomyces cerevisiae strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature controls the culture at 32 DEG C
Be transferred in the eggplant type bottle with culture medium and cultivate 34 hours again after 40 hours, in eggplant type bottle culture medium by mass percentage, by
Following group is grouped as：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.2.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 34 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.5%, yeast extract 0.2%, peptone 0.1%, bean cake powder 0.5%, phosphoric acid
Potassium dihydrogen 0.05%, surplus are water, pH=7.2.
Saccharomyces cerevisiae is seeded in the fermentor containing culture medium by 5% inoculum concentration, under the conditions of 35 DEG C, is passed through nothing
Bacterium air after culture 8 hours, starts to sample microscopy, and incubation time is taken a sample to check secondary for every 4 hours within the scope of 8 to 24 hours,
Incubation time checks primary for every 2 hours within the scope of 24 to 48 hours, it is ensured that without miscellaneous bacteria, after saccharomyces cerevisiae, which sprouts, to be stablized, hair
Ferment culture is completed.
Fermentation liquid is subjected to 8 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 32% is added in liquid, in intake air temperature is 196 DEG C after stirring evenly, air outlet temperature is
80 DEG C, feed rate is spray-dried under conditions of being 12mL/min, obtains S. cervisiae powder, and bacterial content is equal in the bacterium powder
It is 109Cfu/g。
(3) thermophilic sporotrichum bacterium powder is prepared
After thermophilic sporotrichum conidia powder and flour are mixed, it is inoculated on solid medium, at this point, thermophilic sporotrichum
The weight ratio of conidia powder, flour and solid medium is 1:50:30, the solid medium after inoculation is then put into sterile fermentation
It is that 6cm carries out paving disk by laying depth on the koji tray that chamber inner diameter is 1.2 meters, under conditions of ventilation, consolidating after keeping inoculation
The temperature of body culture medium be 30 DEG C, ferment 45 hours, to koji plate fermentation after, through drying and crushing obtain thermophilic sporotrichum bacterium
Powder, bacterial content is 10 in the bacterium powder9Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and jade
Rice flour is 60 in mass ratio:15:Then the epsom salt, 3.5% for being equivalent to mixture weight 0.4% is added in 25 mixing
Ferric sulfate and 0.15% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 60%.
(4) Geotrichum powder is prepared
After the yam flour of geotrichum candidum conidia powder and 100 mesh is mixed, it is inoculated on solid medium, at this point, geotrichum candidum spore
The weight ratio of sub- powder, yam flour and solid medium is 0.015:1.5:2, then the solid medium after inoculation is put into sterile
It is that 6cm carries out paving disk by laying depth, under conditions of ventilation, after keeping inoculation on the koji tray that chamber inner diameter of fermenting is 1.2 meters
Solid medium temperature be 30 DEG C, ferment 45 hours, to koji plate fermentation after, obtain Geotrichum through drying and crushing
Powder, bacterial content is 10 in the bacterium powder9Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and jade
Rice flour is 70 in mass ratio:15:Then the epsom salt, 3.5% for being equivalent to mixture weight 0.4% is added in 15 mixing
Ferric sulfate and 0.15% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 60%.
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
Embodiment 2
Prepare a kind of crop material waste leavening
(1) Brevibacillus laterosporus bacterium powder is prepared
The Brevibacillus laterosporus strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature is controlled 32
DEG C, culture is transferred in the eggplant type bottle with culture medium again after 36 hours and cultivates 34 hours, and culture medium presses quality percentage in eggplant type bottle
Than meter, it is grouped as by following group：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.5.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 35 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.01%, beef extract 0.07%, peptone 0.1%, sodium chloride 0.01% are remaining
Amount is water, pH=7.5.
Brevibacillus laterosporus is seeded in the fermentor containing culture medium by 4% inoculum concentration, under the conditions of 35 DEG C,
It is passed through filtrated air, after culture 8 hours, starts to sample microscopy, incubation time is within the scope of 8 to 24 hours, the inspection of sampling in every 4 hours
It looks into time, incubation time checks primary, it is ensured that within the scope of 24 to 36 hours without miscellaneous bacteria, when Brevibacillus laterosporus bud for every 2 hours
When spore formation rate reaches 90%, fermented and cultured is completed.
Fermentation liquid is subjected to 7 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 26% is added in liquid, in intake air temperature is 198 DEG C after stirring evenly, air outlet temperature is
75 DEG C, feed rate is spray-dried under conditions of being 20mL/min, obtains Brevibacillus laterosporus bacterium powder, bacterium in the bacterium powder
Content is 109Cfu/g。
(1) S. cervisiae powder is prepared
The saccharomyces cerevisiae strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature controls the culture at 35 DEG C
Be transferred in the eggplant type bottle with culture medium and cultivate 32 hours again after 45 hours, in eggplant type bottle culture medium by mass percentage, by
Following group is grouped as：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.5.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 33 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.01%, yeast extract 0.10%, peptone 0.5%, bean cake powder 0.01%, phosphorus
Acid dihydride potassium 0.5%, surplus are water, pH=7.5.
Saccharomyces cerevisiae is seeded in the fermentor containing culture medium by 3% inoculum concentration, under the conditions of 32 DEG C, is passed through nothing
Bacterium air after culture 8 hours, starts to sample microscopy, and incubation time is taken a sample to check secondary for every 4 hours within the scope of 8 to 24 hours,
Incubation time checks primary for every 2 hours within the scope of 24 to 48 hours, it is ensured that without miscellaneous bacteria, after saccharomyces cerevisiae, which sprouts, to be stablized, hair
Ferment culture is completed.
Fermentation liquid is subjected to 7 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 26% is added in liquid, in intake air temperature is 198 DEG C after stirring evenly, air outlet temperature is
75 DEG C, feed rate is spray-dried under conditions of being 20mL/min, obtains S. cervisiae powder, and bacterial content is equal in the bacterium powder
It is 107Cfu/g。
(3) thermophilic sporotrichum bacterium powder is prepared
After thermophilic sporotrichum conidia powder and flour are mixed, it is inoculated on solid medium, at this point, thermophilic sporotrichum
The weight ratio of conidia powder, flour and solid medium is 1:30:10, the solid medium after inoculation is then put into sterile fermentation
It is that 8cm carries out paving disk by laying depth on the koji tray that chamber inner diameter is 1.2 meters, under conditions of ventilation, consolidating after keeping inoculation
The temperature of body culture medium be 30 DEG C, ferment 35 hours, to koji plate fermentation after, through drying and crushing obtain thermophilic sporotrichum bacterium
Powder, bacterial content is 10 in the bacterium powder8Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and jade
Rice flour is 70 in mass ratio:15:Then the sulphur of the epsom salt, 2% that are equivalent to mixture weight 0.5% is added in 15 mixing
Sour iron and 0.2% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 50%.
(4) Geotrichum powder is prepared
After the yam flour of geotrichum candidum conidia powder and 150 mesh is mixed, it is inoculated on solid medium, at this point, geotrichum candidum spore
The weight ratio of sub- powder, yam flour and solid medium is 0.02:1:1, the solid medium after inoculation is then put into sterile hair
It is that 8cm carries out paving disk by laying depth, under conditions of ventilation, after keeping inoculation on the koji tray that ferment chamber inner diameter is 1.2 meters
The temperature of solid medium be 30 DEG C, ferment 35 hours, to koji plate fermentation after, through drying and crushing obtain Geotrichum powder,
Bacterial content is 10 in the bacterium powder7Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and corn
Powder is 60 in mass ratio:15:Then the sulfuric acid of the epsom salt, 2% that are equivalent to mixture weight 0.5% is added in 25 mixing
Iron and 0.2% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 50%.
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
Embodiment 3
Prepare a kind of crop material waste leavening
(1) Brevibacillus laterosporus bacterium powder is prepared
The Brevibacillus laterosporus strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature is controlled 37
DEG C, culture is transferred in the eggplant type bottle with culture medium again after 24 hours and cultivates 36 hours, and culture medium presses quality percentage in eggplant type bottle
Than meter, it is grouped as by following group：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 30 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.07%, beef extract 0.01%, peptone 0.01%, sodium chloride 0.1% are remaining
Amount is water, pH=7.
Brevibacillus laterosporus is seeded in the fermentor containing culture medium by 3% inoculum concentration, under the conditions of 30 DEG C,
It is passed through filtrated air, after culture 8 hours, starts to sample microscopy, incubation time is within the scope of 8 to 24 hours, the inspection of sampling in every 4 hours
It looks into time, incubation time checks primary, it is ensured that within the scope of 24 to 36 hours without miscellaneous bacteria, when Brevibacillus laterosporus bud for every 2 hours
When spore formation rate reaches 92%, fermented and cultured is completed.
Fermentation liquid is subjected to 5 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 20% is added in liquid, in intake air temperature is 200 DEG C after stirring evenly, air outlet temperature is
76 DEG C, feed rate is spray-dried under conditions of being 8mL/min, obtains Brevibacillus laterosporus bacterium powder, bacterium in the bacterium powder
Content is 108Cfu/g。
(1) S. cervisiae powder is prepared
The saccharomyces cerevisiae strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature controls the culture at 32 DEG C
Be transferred in the eggplant type bottle with culture medium and cultivate 36 hours again after 48 hours, in eggplant type bottle culture medium by mass percentage, by
Following group is grouped as：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=7.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 35 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.1%, yeast extract 1.0%, peptone 0.01%, bean cake powder 0.05%, phosphorus
Acid dihydride potassium 1.0%, surplus are water, pH=7.
Saccharomyces cerevisiae is seeded in the fermentor containing culture medium by 2% inoculum concentration, under the conditions of 30 DEG C, is passed through nothing
Bacterium air after culture 8 hours, starts to sample microscopy, and incubation time is taken a sample to check secondary for every 4 hours within the scope of 8 to 24 hours,
Incubation time checks primary for every 2 hours within the scope of 24 to 48 hours, it is ensured that without miscellaneous bacteria, after saccharomyces cerevisiae, which sprouts, to be stablized, hair
Ferment culture is completed.
Fermentation liquid is subjected to 5 times of concentrations by centrifugal separator first, concentrated broth is obtained, then ferments to the concentration
The precipitated calcium carbonate of concentrated broth weight 20% is added in liquid, in intake air temperature is 200 DEG C after stirring evenly, air outlet temperature is
76 DEG C, feed rate is spray-dried under conditions of being 8mL/min, obtains S. cervisiae powder, and bacterial content is equal in the bacterium powder
It is 108Cfu/g。
(3) thermophilic sporotrichum bacterium powder is prepared
After thermophilic sporotrichum conidia powder and flour are mixed, it is inoculated on solid medium, at this point, thermophilic sporotrichum
The weight ratio of conidia powder, flour and solid medium is 1:20:50, the solid medium after inoculation is then put into sterile fermentation
It is that 7cm carries out paving disk by laying depth on the koji tray that chamber inner diameter is 1.2 meters, under conditions of ventilation, consolidating after keeping inoculation
The temperature of body culture medium be 30 DEG C, ferment 50 hours, to koji plate fermentation after, through drying and crushing obtain thermophilic sporotrichum bacterium
Powder, bacterial content is 10 in the bacterium powder7Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and jade
Rice flour is 50 in mass ratio:18:Then the epsom salt, 2.5% for being equivalent to mixture weight 0.3% is added in 25 mixing
Ferric sulfate and 0.1% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 55%.
(4) Geotrichum powder is prepared
After the yam flour of geotrichum candidum conidia powder and 60 mesh is mixed, it is inoculated on solid medium, at this point, geotrichum candidum spore
The weight ratio of powder, yam flour and solid medium is 0.015:2:1.5, the solid medium after inoculation is then put into sterile hair
It is that 7cm carries out paving disk by laying depth, under conditions of ventilation, after keeping inoculation on the koji tray that ferment chamber inner diameter is 1.2 meters
The temperature of solid medium be 30 DEG C, ferment 50 hours, to koji plate fermentation after, through drying and crushing obtain Geotrichum powder,
Bacterial content is 10 in the bacterium powder8Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and corn
Powder is 50 in mass ratio:18:Then the sulphur of the epsom salt, 2.5% that are equivalent to mixture weight 0.3% is added in 25 mixing
Sour iron and 0.1% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 55%.
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
Embodiment 4
Prepare a kind of crop material waste leavening
(1) Brevibacillus laterosporus bacterium powder is prepared
The Brevibacillus laterosporus strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature is controlled 35
DEG C, culture is transferred in the eggplant type bottle with culture medium again after 30 hours and cultivates 30 hours, and culture medium presses quality percentage in eggplant type bottle
Than meter, it is grouped as by following group：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=6.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 30 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 0.1%, beef extract 0.1%, peptone 0.07%, sodium chloride 0.07% are remaining
Amount is water, pH=6.
Brevibacillus laterosporus is seeded in the fermentor containing culture medium by 1% inoculum concentration, under the conditions of 37 DEG C,
It is passed through filtrated air, after culture 8 hours, starts to sample microscopy, incubation time is within the scope of 8 to 24 hours, the inspection of sampling in every 4 hours
It looks into time, incubation time checks primary, it is ensured that within the scope of 24 to 36 hours without miscellaneous bacteria, when Brevibacillus laterosporus bud for every 2 hours
When spore formation rate reaches 94%, fermented and cultured is completed.
Fermentation liquid is subjected to 10 times of concentrations by centrifugal separator first, concentrated broth is obtained, is then sent out to the concentration
In zymotic fluid be added concentrated broth weight 40% precipitated calcium carbonate, after stirring evenly intake air temperature be 195 DEG C, air outlet temperature
It is 78 DEG C, feed rate is spray-dried under conditions of being 5mL/min, acquisition Brevibacillus laterosporus bacterium powder, in the bacterium powder
Bacterial content is 107Cfu/g。
(1) S. cervisiae powder is prepared
The saccharomyces cerevisiae strain of freeze-drying is transferred on test tube slant in gnotobasis, temperature controls the culture at 30 DEG C
Be transferred in the eggplant type bottle with culture medium and cultivate 30 hours again after 36 hours, in eggplant type bottle culture medium by mass percentage, by
Following group is grouped as：Tryptone 1%, yeast extract 0.5%, sodium chloride 0.5%, surplus are water, pH=6.
It is passed through steam into fermentor, opens stirring, 30 points are kept the temperature when culture medium temperature in fermentor reaches 121 DEG C
Clock closes steam after the completion of sterilizing, and opening cooling system makes the temperature of culture medium be down to 32 DEG C, and culture medium presses quality in fermentor
Percentages are grouped as by following group：Glucose 1.0%, yeast extract 0.01%, peptone 1.0%, bean cake powder 1.0%, phosphoric acid
Potassium dihydrogen 0.01%, surplus are water, pH=6.
Saccharomyces cerevisiae is seeded in the fermentor containing culture medium by 1% inoculum concentration, under the conditions of 37 DEG C, is passed through nothing
Bacterium air after culture 8 hours, starts to sample microscopy, and incubation time is taken a sample to check secondary for every 4 hours within the scope of 8 to 24 hours,
Incubation time checks primary for every 2 hours within the scope of 24 to 48 hours, it is ensured that without miscellaneous bacteria, after saccharomyces cerevisiae, which sprouts, to be stablized, hair
Ferment culture is completed.
Fermentation liquid is subjected to 10 times of concentrations by centrifugal separator first, concentrated broth is obtained, is then sent out to the concentration
In zymotic fluid be added concentrated broth weight 40% precipitated calcium carbonate, after stirring evenly intake air temperature be 195 DEG C, air outlet temperature
It is 78 DEG C, feed rate is spray-dried under conditions of being 5mL/min, obtains S. cervisiae powder, bacterial content in the bacterium powder
It is 107Cfu/g。
(3) thermophilic sporotrichum bacterium powder is prepared
After thermophilic sporotrichum conidia powder and flour are mixed, it is inoculated on solid medium, at this point, thermophilic sporotrichum
The weight ratio of conidia powder, flour and solid medium is 1:10:20, the solid medium after inoculation is then put into sterile fermentation
It is that 10cm carries out paving disk by laying depth, under conditions of ventilation, after keeping inoculation on the koji tray that chamber inner diameter is 1.2 meters
The temperature of solid medium be 30 DEG C, ferment 40 hours, to koji plate fermentation after, obtain thermophilic sporotrichum through drying and crushing
Bacterium powder, bacterial content is 10 in the bacterium powder8Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and
Corn flour is 50 in mass ratio:20:Then the epsom salt, 3% for being equivalent to mixture weight 0.3% is added in 25 mixing
Ferric sulfate and 0.1% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 55%.
(4) Geotrichum powder is prepared
After the yam flour of geotrichum candidum conidia powder and 200 mesh is mixed, it is inoculated on solid medium, at this point, geotrichum candidum spore
The weight ratio of sub- powder, yam flour and solid medium is 0.01:2:2, the solid medium after inoculation is then put into sterile hair
It is that 10cm carries out paving disk by laying depth, under conditions of ventilation, after keeping inoculation on the koji tray that ferment chamber inner diameter is 1.2 meters
Solid medium temperature be 30 DEG C, ferment 40 hours, to koji plate fermentation after, obtain Geotrichum through drying and crushing
Powder, bacterial content is 10 in the bacterium powder8Cfu/g.Wherein, solid medium is prepared by following methods：By wheat bran, bean cake powder and jade
Rice flour is 50 in mass ratio:20:Then the sulphur of the epsom salt, 3% that are equivalent to mixture weight 0.3% is added in 25 mixing
Sour iron and 0.1% potassium dihydrogen phosphate, finally add water to biodiversity percentage be 55%.
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
The leavening prepared in embodiment 1 to embodiment 4 is used for the fermentation of crop material waste, verifying above-mentioned 4 respectively
Middle leavening is to the ferment effect of crop material waste, wherein the results are shown in Table 1 leavening is not added as control.
As shown in Table 1, be not used the present invention in leavening control group begin with the wine flavour time be the 49th day, completely send out
The time of ferment maturation is the 69th day, and organic acid content is 7.21% after fermentation, and has used in the present invention and opened after leavening
There is wine flavour time advance in beginning 36-41 days, and the time advance of complete fermentation maturation 49-54 days, organic acid after fermentation
Content improves 4.45-10.02%, it is sufficient to prove that leavening there is excellent fermentation to imitate crop material waste in the present invention
Fruit is not only effectively shortened fermentation time, coordinates the nutrients ratio in finally obtained tunning more.
Finally, it is stated that preferred embodiment above is only used to illustrate the technical scheme of the present invention and not to limit it, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (10)
1. a kind of crop material waste leavening, which is characterized in that the leavening is by Brevibacillus laterosporus bacterium powder, wine brewing
Saccharomycete powder, thermophilic sporotrichum bacterium powder and Geotrichum powder form, and the bacteria concentration of each bacterium is 10 in the leavening7-
109Cfu/g；The Geotrichum powder is made by following methods：After geotrichum candidum conidia powder and yam flour are mixed, it is inoculated into solid
On culture medium, the solid medium after inoculation is then subjected to koji plate fermentation, after the koji plate fermentation, is obtained through drying and crushing
Obtain Geotrichum powder.
2. a kind of crop material waste leavening as described in claim 1, which is characterized in that the geotrichum candidum conidia powder,
The weight ratio of yam flour and solid medium is 0.01-0.02:1-2:1-2.
3. a kind of crop material waste leavening as claimed in claim 2, which is characterized in that the solid medium by with
The preparation of lower section method：It is in mass ratio 50-70 by wheat bran, bean cake powder and corn flour:15-20:Then 15-25 mixing is added suitable
In the epsom salt of mixture weight 0.3-0.5%, the potassium dihydrogen phosphate of the ferric sulfate of 2-3.5% and 0.1-0.2%, finally
Adding water to biodiversity percentage is 50-60%.
4. a kind of described in any item preparation methods of crop material waste leavening of claim 1-3, which is characterized in that institute
The method of stating includes the following steps：
(1) Brevibacillus laterosporus is seeded in the fermentor containing culture medium and carries out fermented and cultured, the fermented and cultured knot
Fermentation liquid is concentrated first by Shu Hou, obtains concentrated broth, and lightweight carbonic acid is then added into the concentrated broth
Calcium is spray-dried after stirring evenly, and obtains Brevibacillus laterosporus bacterium powder；
(2) saccharomyces cerevisiae is seeded in the fermentor containing culture medium and carries out fermented and cultured, it is first after the fermented and cultured
First fermentation liquid is concentrated, concentrated broth is obtained, precipitated calcium carbonate then is added into the concentrated broth, after stirring evenly
It is spray-dried, obtain S. cervisiae powder；
(3) it after mixing thermophilic sporotrichum conidia powder and flour, is inoculated on solid medium, then by the solid after inoculation
Culture medium carries out koji plate fermentation, after the koji plate fermentation, obtains thermophilic sporotrichum bacterium powder through drying and crushing；
(4) it after mixing geotrichum candidum conidia powder and yam flour, is inoculated on solid medium, then by the solid culture after inoculation
Base carries out koji plate fermentation, after the koji plate fermentation, obtains Geotrichum powder through drying and crushing；
(5) after mixing by bacterium powder each in step (1)-(4), crop material waste leavening is made.
5. method as claimed in claim 4, which is characterized in that in step (1) and step (2), the concentration is specially to be concentrated
5-10 times.
6. method as claimed in claim 4, which is characterized in that in step (1) and step (2), the precipitated calcium carbonate adds
Enter the 20-40% that amount is the concentrated broth weight.
7. method as claimed in claim 4, which is characterized in that in step (1) and step (2), when the spray drying, air inlet
Mouth temperature is 195-200 DEG C, and air outlet temperature is 75-80 DEG C, feed rate 5-20mL/min.
8. method as claimed in claim 4, which is characterized in that in step (3), the thermophilic sporotrichum conidia powder, flour
Weight ratio with solid medium is 1:10-50:10-50.
9. method as claimed in claim 4, which is characterized in that in step (3) and step (4), the solid medium by with
The preparation of lower section method：It is in mass ratio 50-70 by wheat bran, bean cake powder and corn flour:15-20:Then 15-25 mixing is added suitable
In the epsom salt of mixture weight 0.3-0.5%, the potassium dihydrogen phosphate of the ferric sulfate of 2-3.5% and 0.1-0.2%, finally
Adding water to biodiversity percentage is 50-60%.
10. a kind of described in any item crop material waste leavenings of claim 1-3 are preparing the application in biological feedstuff.
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