CN110384175A - The method of yeast culture and the application of yeast culture are prepared using vinasse - Google Patents
The method of yeast culture and the application of yeast culture are prepared using vinasse Download PDFInfo
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Abstract
The invention discloses a kind of applications of method and yeast culture that yeast culture is prepared using vinasse, method is the following steps are included: S1: the supplementary carbon source into vinasse, obtain pretreatment of distilled grain, it is inoculated with mixed bacteria liquid into gained pretreatment of distilled grain and mixed enzyme solution is added, fermentation base-material, fermented and cultured 96-120h is made；S2: being inoculated with bacillus subtilis after fermentation, and after continuing fermented and cultured for 24 hours, drying pulverizes and sieves, and obtains yeast culture；It wherein, include 3 saccharomyces cerevisiae, lactic acid bacteria and aspergillus niger strains in the mixed bacteria liquid；It include α-amylase, carbohydrase, phytase and protease in the mixed enzyme solution.The present invention, by the addition heap fermentation of Mixed Microbes and mixed enzyme solution, obtains yeast culture, vinasse is turned waste into wealth using vinasse as matrix, and provides the feed of rich in nutrition content for animal, conducive to the immunity for improving animal.
Description
Technical field
The present invention relates to technical field of animal, in particular to it is a kind of using vinasse prepare yeast culture method and
The application of yeast culture.
Background technique
Yeast culture refers to that saccharomycete is obtained in defined medium by depth fermentation under certain process conditions
Complicated tunning, it includes products of cellular metabolism (containing polypeptide, organic acid, amino acid, nucleic acid, enzyme, unknown growth factor
Deng), denaturation culture medium (containing oligosaccharides, polypeptide etc.) and yeast cells itself (protein, amino acid, nucleic acid, cell wall polysaccharides etc.),
It is full of nutrition, it is a kind of novel green probiotics.Yeast culture can promote the growth of animal gastrointestinal microbiological, improvement to disappear
Change ability improves immunity, detoxification, therefore is conducive to the healthy growth of animal, has a extensive future in Production of Livestock and Poultry.
Vinasse, alias red wine dregs, fermented grain grain, dregs of rice etc. are remaining residues after the wine brewing such as rice, wheat, jowar.Crude protein content
It can reach 25% or so, it be one as the feed of ox or other animals and is selected well.But ox directly is fed with vinasse
Or not only nutritive value is not fully utilized other animals, and mouthfeel is also poor.In addition, also containing a large amount of fiber in vinasse
Element, fragrance matter etc., currently, vinasse main application is directly as feed, production flavouring and culture edible mushroom etc., added value
It is low.Therefore, how the vinasse that pile up like a mountain to be turned waste into wealth by microorganism channel, to be prepared into the feed of suitable feeding animals
It is urgently to be resolved.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide the methods for preparing yeast culture using vinasse, using vinasse as matrix, lead to
The addition heap fermentation for crossing Mixed Microbes and mixed enzyme solution sufficiently decomposes macromolecular substances in vinasse, adds bacillus subtilis
Bacterium, while increasing probiotics, moreover it is possible to yeast cells self-dissolving be promoted by its metabolite, conducive to more yeast trainings are obtained
Support object.
It is a still further object of the present invention to provide a kind of yeast cultures prepared using the method in animal
It applies, vinasse can be turned waste into wealth in feed, and provide the feed of rich in nutrition content for animal, to improve the immune of animal
Power.
In order to realize object of the present invention and further advantage, provides and a kind of prepare yeast culture using vinasse
Method, comprising the following steps:
S1: the supplementary carbon source into vinasse obtains pretreatment of distilled grain, and mixed bacteria liquid is inoculated with into gained pretreatment of distilled grain and is added
Enter mixed enzyme solution, fermentation base-material, fermented and cultured 96-120h is made；
S2: being inoculated with bacillus subtilis after fermentation, and after continuing fermented and cultured for 24 hours, drying pulverizes and sieves, and obtains
Yeast culture；
It wherein, include 3 saccharomyces cerevisiae, lactic acid bacteria and aspergillus niger strains in the mixed bacteria liquid；The mixed enzyme solution
In include α-amylase, carbohydrase, phytase and protease.
Preferably, the carbon source is corn flour；
It is mixed with the corn flour by weight 1:0.1-0.6 after the vinasse are dry.
Preferably, after each strain in the Mixed Microbes is fermented respectively, saccharomyces cerevisiae bacterium solution, lactic acid are respectively obtained
Bacterium bacterium solution and aspergillus niger bacterium solution, then by the saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution by volume 1:
1-3:1-3 mixing, obtains mixed bacteria liquid；
By pretreatment of distilled grain total weight, the mixed bacteria liquid inoculum concentration is 5-20%；The viable count of the mixed bacteria liquid
It is 1.0 × 108-1.0×109cfu/mL。
Preferably, the mixed enzyme solution is by liquefaction enzyme solutions, Glucoamylase Solution, phytase solution, protein enzyme solution
1:0.4-0.8:0.1-0.2:0.2-1 is mixed by volume, wherein liquefaction enzyme solutions, Glucoamylase Solution, phytase are molten
Liquid, protein enzyme solution enzyme activity be 10000U/mL；
By the pretreatment of distilled grain total weight, the mixed enzyme solution adds 1-3%.
Preferably, fermentation condition in S1 are as follows: the fermentation base-material tiles, at 30-35 DEG C of temperature, every 5-6h stirring
Once, primary every 2-3h stirring when temperature rises to 40 DEG C or more；
The thickness of the fermentation base-material tiling specifically: when fermentation temperature is lower than 40 DEG C, tile with a thickness of 40-60cm；
After fermentation temperature rises to 40 DEG C and 40 DEG C, tile with a thickness of 20-40cm.
Preferably, S1 is further comprising the steps of:
When the base-material that ferments ferments to 72-96h, a yeast powder is supplemented；It is described by the pretreatment of distilled grain total weight
Yeast powder supplements 4-10%.
Preferably, by the pretreatment of distilled grain total weight, the bacillus subtilis inoculum concentration is 1.0 ×
106cfu/g。
Preferably, further includes:
The fermentation medium of the saccharomyces cerevisiae comprises the following components in parts by weight: 1-3 parts of yeast extract, 2-5 parts of peptone,
2-5 parts of glucose and distilled water 1000mL；Fermentation temperature: 27-30 DEG C, fermentation condition: 180-200r/min ferments 3-5 days；
The fermentation medium of the aspergillus niger comprises the following components in parts by weight: 1-3 parts of yeast extract, 2-5 parts of peptone, Portugal
2-5 parts of grape sugar and distilled water 1000mL；Fermentation temperature: 27-30 DEG C, fermentation condition: 180-200r/min ferments 3-5 days；
The fermentation medium of the lactic acid bacteria comprises the following components in parts by weight: 10-13 parts of peptone, beef extract 10-12
Part, 5-8 parts of yeast extract, 2-3 parts of diammonium hydrogen citrate, 20-25 parts of glucose, 0.5-1 parts of Tween 80,5-8 parts of sodium acetate, phosphorus
2-5 parts of sour hydrogen dipotassium, 0.5-1 parts of magnesium sulfate, 0.1-0.3 parts of manganese sulfate and distilled water 1000mL, pH6.2-6.6；Fermentation temperature
Degree: 35-37 DEG C；Fermentation condition: stationary culture 3-5 days；
The fermentation medium of the bacillus subtilis comprises the following components in parts by weight: 15-20 parts of glucose, dregs of beans
30-35 parts, 10-15 parts of starch, 0.1-0.3 parts of manganese sulfate, 3-5 parts of dipotassium hydrogen phosphate, 2-3 parts of potassium dihydrogen phosphate, magnesium sulfate
0.05-0.1 parts, 2-5 parts of yeast extract, 1-2 parts of iron chloride, 1-3 parts of calcium carbonate and distilled water 1000mL, PH7.0-7.2；Fermentation
Temperature: 35-37 DEG C；Fermentation condition: 200-280r/min ferments 3-5 days.
Preferably, it after adding bacillus subtilis in S2, tiles and ferments for the thickness of 100cm, after obtaining self-dissolving
Material；
Material after self-dissolving is dried to water content 10% at 85-90 DEG C hereinafter, crushing and crossing 40 meshes, obtains the yeast
Culture.
The present invention also provides a kind of yeast cultures prepared using the method to apply in animal feed.
The present invention is include at least the following beneficial effects:
The present invention using vinasse as matrix, due to vinasse be by grain after everfermentation remaining residue, carbon in residue
Source content is relatively low, if directly as fermentation base-material using the existence that will be unfavorable for strain, therefore the present invention passes through elder generation
After supplementing certain carbon source into vinasse, pretreatment of distilled grain is obtained, then using the various nutriments in pretreatment of distilled grain, is led to
Cross addition the mixed bacteria liquid containing saccharomyces cerevisiae, lactic acid bacteria and aspergillus niger, and containing α-amylase, carbohydrase, phytase with
And the mixed bacteria liquid of protease, fully degraded is carried out, specifically, due to the starch, protein, rice husk etc. that are participated in vinasse, it is black
Aspergillus contains the almost a full set of catabolic enzyme for botany raw material, including amylase, lipase, protease, cellulase, wood
Dextranase, dextranase, mannase etc. is able to carry out the decomposition of total synergy to vegetable raw material, and yeast, lactic acid bacteria
Etc. shortages using the ability of starch, macro-molecular protein, xylan, cellulose, glucan etc., the raw material containing mentioned component is not
Saccharomycete and lactic acid bacteria can be efficiently produced, and after the serial enzymatic treatment that aspergillus niger generates, starch is broken down into fermentable
Glucose, protein are broken down into the peptide or amino acid of small molecule, and xylan is broken down into xylose, the quilts such as glucan, cellulose
It is decomposed into glucose monomer, becomes saccharomycete and lactic acid bacteria is easy the nutrition of sorption enhanced, carry out in conjunction with mixed enzyme agent to wine
After poor raw material carries out preliminary decomposition, the coordination process that would be even more beneficial to above-mentioned strain is carried out, the yeast culture prepared with this
It is used directly as animal feed, the trophic level and digestibility of fermented feed can be greatly improved.
It is preferably to be used directly as animal feed in order to guarantee tunning small molecule nutritional ingredient rich in,
Through mixed fermentation for a period of time after, then add a certain amount of bacillus subtilis, on the one hand increase probiotics, on the other hand
It can also promote the self-dissolving of yeast cells by the metabolite of producing bacillus subtilis life, such as enzyme, generate more yeast
Culture.The present invention changes the conventional yeast culture for preparing and consumes a large amount of molasses, glucose, dregs of beans, urea, bicarbonate
The problem of carbon sources such as ammonium, nitrogen source, and a small amount of carbon source is only required supplementation with, it economizes on resources, also greatly reduces production cost.
In addition, the application the invention also discloses prepared yeast culture in animal feed, mainly passes through this
The yeast product nutritional ingredient rich in of the method preparation of invention, through strain degradation is suitable by the nutriment in vinasse
The directly edible small-molecule substance of suitable animal, changes palatability of the vinasse as feed, improves the utilization rate of nutriment,
A large amount of vinasse can also be turned waste into wealth；Yeast culture of the invention contains a large amount of saccharomyces cerevisiae, it can reach 2.05 ×
109Cfu/g can improve the immunity of animal directly as animal feed feeding animals.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Specific embodiment
The present invention is described in further detail below, to enable those skilled in the art's refer to the instruction text being capable of evidence
To implement.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded one or more
The presence or addition of a other elements or combinations thereof.
Saccharomyces cerevisiae used in the present invention is bought in Angel Yeast company；Lactic acid bacteria, black-koji mould and bacillus subtilis
Bacterium is provided by Hua Zhong Agriculture University's Fermentation Engineering room.
Embodiment 1
The preparation of distiller's dried grain: the fresh spirit stillage transported from brewery is dried to moisture content at 115 DEG C with dryer
Lower than 15%, distiller's dried grain is obtained.
The preparation of saccharomyces cerevisiae bacterium solution: 30 DEG C of activation 20h of YPD slant medium are first used, then with YPD fermentation medium 30
DEG C, fermented and cultured 5 days under the conditions of 180r/min.
YPD slant medium comprises the following components in parts by weight: 3 parts of yeast extract, 4 parts of peptone, 4 parts of glucose and steaming
Distilled water 1000mL；
YPD fermentation medium comprises the following components in parts by weight: 2 parts of yeast extract, 3 parts of peptone, 3 parts of glucose and steaming
Distilled water 1000mL.
The preparation of aspergillus niger bacterium solution: 30 DEG C of activation 20h of YPD slant medium are first used, then with YPD fermentation medium 30
DEG C, fermented and cultured 4 days under the conditions of 180r/min.
YPD slant medium comprises the following components in parts by weight: 3 parts of yeast extract, 5 parts of peptone, 5 parts of glucose and steaming
Distilled water 1000mL；
YPD fermentation medium comprises the following components in parts by weight: 2 parts of yeast extract, 3 parts of peptone, 3 parts of glucose and steaming
Distilled water 1000mL.
The preparation of lactic acid bacterial liquid: 35 DEG C of activation 20h of MRS slant medium are first used, then with MRS fermentation medium 37
DEG C, standing for fermentation culture 5 days.
MRS slant medium comprises the following components in parts by weight: 13 parts of peptone, 12 parts of beef extract, 8 parts of yeast extract, lemon
3 parts of lemon acid hydrogen diammonium, 25 parts of glucose, 1 part of Tween 80,8 parts of sodium acetate, 5 parts of dipotassium hydrogen phosphate, 1 part of magnesium sulfate, manganese sulfate
0.3 part and distilled water 1000mL, pH6.6；
MRS fermentation medium comprises the following components in parts by weight: 12 parts of peptone, 10 parts of beef extract, 6 parts of yeast extract, lemon
2 parts of lemon acid hydrogen diammonium, 20 parts of glucose, 0.5 part of Tween 80,6 parts of sodium acetate, 3 parts of dipotassium hydrogen phosphate, 0.5 part of magnesium sulfate, sulphur
0.1 part of sour manganese and distilled water 1000mL, pH6.6.
The preparation of bacillus subtilis bacterium solution: 35 DEG C of activation 20h of slant medium are first used, then with fermentation medium 37
DEG C, under the conditions of 200r/min, fermented and cultured 5 days.
Wherein, slant medium comprises the following components in parts by weight: 20 parts of glucose, 35 parts of dregs of beans, 15 parts of starch, sulfuric acid
0.3 part of manganese, 5 parts of dipotassium hydrogen phosphate, 3 parts of potassium dihydrogen phosphate, 0.1 part of magnesium sulfate, 5 parts of yeast extract, 2 parts of iron chloride, 3 parts of calcium carbonate
And distilled water 1000mL, PH7.0；
Fermentation medium comprises the following components in parts by weight: 15 parts of glucose, 30 parts of dregs of beans, 12 parts of starch, manganese sulfate
0.12 part, 4 parts of dipotassium hydrogen phosphate, 2 parts of potassium dihydrogen phosphate, 0.05 part of magnesium sulfate, 2 parts of yeast extract, 1 part of iron chloride, 2 parts of calcium carbonate
And distilled water 1000mL, PH7.0.
Embodiment 2
Using the method for vinasse fermentation yeast culture, include the following steps:
Distiller's dried grain 200kg in Example 1, after distiller's dried grain is mixed with corn flour by weight 1:0.2, by distiller's dried grain
The mixture of distiller's dried grain and corn flour and water are stirred and evenly mixed with material-water ratio 1:1 with the total amount meter of corn flour, and adjusts pH and is
5.0 or so, obtain pretreatment of distilled grain；
Based on the butt total amount of pretreatment of distilled grain, 5% mixed bacteria liquid (saccharomyces cerevisiae bacterium solution, lactic acid in embodiment 1 are accessed
1:1:1 is mixed by volume for bacterium bacterium solution and aspergillus niger bacterium solution), and mixed enzyme solution 1% is added (liquefaction enzyme solutions, carbohydrase are molten
1:0.4:0.1:0.2 is mixed by volume for liquid, phytase solution, protein enzyme solution), fermentation base-material is obtained, when 72h is sent out
Add 5% yeast extract in fermentation base-material after ferment, before fermentation temperature reaches 40 DEG C, the initial material heap of fermenting product with a thickness of
50cm, while when being kept for 30-35 DEG C of fermentation temperature, it is primary every 5h stirring；When fermentation temperature reaches 40 DEG C and 40 DEG C or more, heap
Body thickness drops to 10cm, and keeps primary every 2-3h stirring.
It is inoculated with 1.0 × 10 after fermentation6The bacillus subtilis of cfu/g, heap thickness 100cm, accumulation for 24 hours, are dried at 85 DEG C
It does to water content 10% hereinafter, crush and cross 40 meshes, obtains yeast culture.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure yeast number be 1.9 ×
109cfu/g；Total protein concentration is 15%.
Embodiment 3
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 2 is: by weight the mixing of 1:0.3, remaining step is homogeneous for distiller's dried grain and corn flour
Together.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.93 × 109cfu/g；Total protein concentration is 16.7%.
Embodiment 4
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 2 is: by weight the mixing of 1:0.4, remaining step is homogeneous for distiller's dried grain and corn flour
Together.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.95 × 109cfu/g；Total protein concentration is 17.7%.
Embodiment 5
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 2 is: by weight the mixing of 1:0.5, remaining step is homogeneous for distiller's dried grain and corn flour
Together.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.93 × 109cfu/g；Total protein concentration is 18.3%.
Embodiment 6
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 5 is: Mixed Microbes (press by saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution
Volume ratio 1:2:2 mixing) inoculum concentration be 5%, remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.96 × 109cfu/g；Total protein concentration is 19%.
Embodiment 7
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 5 is: Mixed Microbes (press by saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution
Volume ratio 1:2:2 mixing) inoculum concentration be 7%, remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.98 × 109cfu/g；Total protein concentration is 19.5%.
Embodiment 8
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 5 is: Mixed Microbes (press by saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution
Volume ratio 1:2:2 mixing) inoculum concentration be 10%, remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 2.0 × 109cfu/g；Total protein concentration is 19.9%.
Embodiment 9
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 8 is: mixed enzyme solution 2% is added, and (liquefaction enzyme solutions, Glucoamylase Solution, phytase are molten
1:0.6:0.2:1 is mixed by volume for liquid, protein enzyme solution) remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 2.03 × 109cfu/g；Total protein concentration is 22%.
Embodiment 10
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 8 is: mixed enzyme solution 3% is added, and (liquefaction enzyme solutions, Glucoamylase Solution, phytase are molten
1:0.6:0.2:1 is mixed by volume for liquid, protein enzyme solution) remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 2.0 × 109cfu/g；Total protein concentration is 25%.
Embodiment 11
Using the method for vinasse fermentation yeast culture, include the following steps:
Difference from Example 10 is: Mixed Microbes (press by saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution
Volume ratio 1:3:3 mixing) inoculum concentration is 10%, mixed enzyme solution 1% is added, and (liquefaction enzyme solutions, Glucoamylase Solution, phytase are molten
1:0.8:0.2:1 is mixed by volume for liquid, protein enzyme solution), yeast powder supplement 8%, remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 2.05 × 109cfu/g；Total protein concentration is 26%.
Embodiment 12
Using the method for vinasse fermentation yeast culture, include the following steps:
With embodiment 11 the difference is that: Mixed Microbes (press by saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution
Volume ratio 1:3:3 mixing) inoculum concentration be 20%, yeast powder supplement 10%, remaining step is all the same.
It is above-mentioned, before being inoculated with bacillus subtilis after fermentation, using PCA counting method, measure gained yeast culture
Yeast number is 1.93 × 109cfu/g；Total protein concentration is 25.4%.
Embodiment 2- embodiment 12 after mixed bacteria heap fermentation, produces a large amount of ferment compared with prior art
Mother provides basis to provide a large amount of yeast culture, then by the addition of bacillus subtilis, heap fermentation, heating is favorably
In the secondary metabolites raw by producing bacillus subtilis, the allogenic material for promoting yeast cells self-dissolving is provided, in favor of obtaining
Small molecule is easy to the yeast culture of animal absorption.
Embodiment 13
Shijiazhuang City of Hebei Province pig farm, yeast culture prepared by embodiment 11 are used directly as feed, simultaneously
It is 50 jin and identical 30 pigs of age number by weight, is divided into A group, B group, C group, wherein A group feeds the yeast training of embodiment 11
Object is supported, B group feeds the feed of market purchase, and C group does not feed any feed, and A group and B group are to feed 2 daily on the basis of daily ration
Jin feed, C group are adding 2 jin on the basis of basal diet, as a result, it has been found that: compared with C group, A group is delivered for sale 20 days in advance, B group 5 days；
Before delivering for sale, C group has 3 viral infection symptoms occur, restores normal through treatment 2,1 death；B group 2 appearance
Viral infection symptoms restore normal through treatment 1 but feed intake are decreased obviously, 1 death, while period, dysentery occur there are also 1
Disease restores normal through treatment；There is not any illness in 10 pigs of A group.
Embodiment 14
Xinxiang City, Henan Province chicken house, yeast culture prepared by embodiment 11 are used directly as feed, simultaneously will
Identical 300 broiler chicken of weight, 1 age in days number, are divided into A group, B group, and wherein A group feeds the yeast culture of embodiment 11, B group
The feed of market purchase is fed, is observed 20 days, A group and B group feed same amount of feed daily, as a result, it has been found that the chicken A group in 20 ages
0.3kg in average weight ratio B group, and A group does not occur any illness, and B group has 20 dysentery phenomenon occur, restores disease through treatment
Shape, it is seen then that the yeast culture of the application preparation is used directly as feed, can be significantly improved immunity, is conducive to the life of chicken
It is long.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
Showing in specific details and here.
Claims (10)
1. a kind of method for preparing yeast culture using vinasse, which comprises the following steps:
S1: the supplementary carbon source into vinasse obtains pretreatment of distilled grain, and mixed bacteria liquid is inoculated with into gained pretreatment of distilled grain and is added mixed
Fermentation base-material, fermented and cultured 96-120h is made in synthase liquid；
S2: being inoculated with bacillus subtilis after fermentation, and after continuing fermented and cultured for 24 hours, drying pulverizes and sieves, and obtains yeast
Culture；
It wherein, include 3 saccharomyces cerevisiae, lactic acid bacteria and aspergillus niger strains in the mixed bacteria liquid；It is wrapped in the mixed enzyme solution
Include α-amylase, carbohydrase, phytase and protease.
2. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that the carbon source is corn
Powder；
It is mixed with the corn flour by weight 1:0.1-0.6 after the vinasse are dry.
3. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that in the Mixed Microbes
After each strain is fermented respectively, saccharomyces cerevisiae bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution are respectively obtained, then by the wine
1:1-3:1-3 is mixed by volume for brewer yeast bacterium solution, lactic acid bacterial liquid and aspergillus niger bacterium solution, obtains mixed bacteria liquid；
By pretreatment of distilled grain total weight, the mixed bacteria liquid inoculum concentration is 5-20%；The viable count of the mixed bacteria liquid is 1.0
×108-1.0×109cfu/mL。
4. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that the mixed enzyme solution is
By liquefaction enzyme solutions, Glucoamylase Solution, phytase solution, protein enzyme solution 1:0.4-0.8:0.1-0.2:0.2-1 by volume
Mix, wherein liquefaction enzyme solutions, Glucoamylase Solution, phytase solution, protein enzyme solution enzyme activity be 10000U/mL；
By the pretreatment of distilled grain total weight, the mixed enzyme solution adds 1-3%.
5. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that fermentation condition in S1
Are as follows: the fermentation base-material is tiled, it is primary every 5-6h stirring at 30-35 DEG C of temperature, when temperature rises to 40 DEG C or more, often
It is primary every 2-3h stirring；
The thickness of the fermentation base-material tiling specifically: when fermentation temperature is lower than 40 DEG C, tile with a thickness of 40-60cm；Work as hair
Ferment temperature rises to 40 DEG C and 40 DEG C or more, tiles with a thickness of 20-40cm.
6. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that S1 further includes following step
It is rapid:
When the base-material that ferments ferments to 72-96h, a yeast powder is supplemented；By the pretreatment of distilled grain total weight, the yeast
Powder supplements 4-10%.
7. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that press the pretreatment liquor
Poor total weight, the bacillus subtilis inoculum concentration are 1.0 × 106cfu/g。
8. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that further include:
The fermentation medium of the saccharomyces cerevisiae comprises the following components in parts by weight: 1-3 parts of yeast extract, 2-5 parts of peptone, grape
Sugared 2-5 parts and distilled water 1000mL；Fermentation temperature: 27-30 DEG C, fermentation condition: 180-200r/min ferments 3-5 days；
The fermentation medium of the aspergillus niger comprises the following components in parts by weight: 1-3 parts of yeast extract, 2-5 parts of peptone, glucose
2-5 parts and distilled water 1000mL；Fermentation temperature: 27-30 DEG C, fermentation condition: 180-200r/min ferments 3-5 days；
The fermentation medium of the lactic acid bacteria comprises the following components in parts by weight: 10-13 parts of peptone, 10-12 parts of beef extract, ferment
Female cream 5-8 parts, 2-3 parts of diammonium hydrogen citrate, 20-25 parts of glucose, 0.5-1 parts of Tween 80,5-8 parts of sodium acetate, phosphoric acid hydrogen two
2-5 parts of potassium, 0.5-1 parts of magnesium sulfate, 0.1-0.3 parts of manganese sulfate and distilled water 1000mL, pH6.2-6.6；Fermentation temperature: 35-
37℃；Fermentation condition: stationary culture 3-5 days；
The fermentation medium of the bacillus subtilis comprises the following components in parts by weight: 15-20 parts of glucose, dregs of beans 30-35
Part, 10-15 parts of starch, 0.1-0.3 parts of manganese sulfate, 3-5 parts of dipotassium hydrogen phosphate, 2-3 parts of potassium dihydrogen phosphate, magnesium sulfate 0.05-0.1
Part, 2-5 parts of yeast extract, 1-2 parts of iron chloride, 1-3 parts of calcium carbonate and distilled water 1000mL, PH7.0-7.2；Fermentation temperature: 35-
37℃；Fermentation condition: 200-280r/min ferments 3-5 days.
9. the method for preparing yeast culture using vinasse as described in claim 1, which is characterized in that add withered grass bud in S2
After spore bacillus, tiles and ferment for the thickness of 100cm, the material after obtaining self-dissolving；
Material after self-dissolving is dried to water content 10% at 85-90 DEG C hereinafter, crushing and crossing 40 meshes, obtains the Yeast Cultivation
Object.
10. a kind of yeast culture using the described in any item method preparations of claim 1-9 is applied in animal feed.
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| CN111513191A (en) * | 2020-04-24 | 2020-08-11 | 路德环境科技股份有限公司 | Fermentation method of yeast culture with Maotai-flavor distiller's grains as raw material |
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| CN114982864A (en) * | 2022-05-13 | 2022-09-02 | 贵州大学 | Solid-state fermentation distiller's grains and preparation method and application thereof |
| CN115624091A (en) * | 2022-10-26 | 2023-01-20 | 宜宾市南溪区国科中农生物科技有限公司 | White spirit vinasse feed and preparation method thereof |
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