CN110384175B - Method for preparing yeast culture from distiller's grains and application of yeast culture - Google Patents
Method for preparing yeast culture from distiller's grains and application of yeast culture Download PDFInfo
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Abstract
The invention discloses a method for preparing a yeast culture by using vinasse and application of the yeast culture, wherein the method comprises the following steps of: s1: supplementing a carbon source to the vinasse to obtain a pre-treated vinasse, inoculating a mixed bacterial liquid to the obtained pre-treated vinasse, adding a mixed enzyme liquid to prepare a fermentation base material, and performing fermentation culture for 96-120 h; s2: inoculating bacillus subtilis after fermentation is finished, continuing fermentation culture for 24 hours, drying, crushing and sieving to obtain a yeast culture; wherein the mixed bacterial liquid comprises 3 strains of saccharomyces cerevisiae, lactobacillus and aspergillus niger; the mixed enzyme liquid comprises liquefying enzyme, saccharifying enzyme, phytase and protease. The yeast culture is obtained by adding the mixed bacteria and the mixed enzyme liquid into the vinasse serving as the matrix and stacking and fermenting the vinasse, so that the vinasse is changed into valuable, the feed with rich nutrient components is provided for animals, and the immunity of the animals is improved.
Description
Technical Field
The invention relates to the technical field of animal feed, in particular to a method for preparing a yeast culture by using vinasse and application of the yeast culture.
Background
The yeast culture is a complex fermentation product obtained by deep fermentation of yeast in a specific culture medium under certain process conditions, and comprises cell metabolites (containing polypeptide, organic acid, amino acid, nucleic acid, enzymes, unknown growth factors and the like), a modified culture medium (containing oligosaccharide, polypeptide and the like) and yeast cells (protein, amino acid, nucleic acid, cell wall polysaccharide and the like), is rich in nutrition, and is a novel green microecological preparation. The yeast culture can promote the growth of microorganisms in intestines and stomach of animals, improve the digestion capability, improve the immunity and detoxify, thereby being beneficial to the healthy growth of the animals and having wide application prospect in the production of livestock and poultry.
Distiller's grains, namely red distiller's grains, fermented grains, dregs and the like, are residues left after brewing rice, wheat, sorghum and the like. The content of crude protein can reach about 25 percent, and the crude protein is a good choice when being used as feed of cattle or other animals. However, feeding the cattle or other animals directly with the distiller's grains not only can not make full use of the nutritive value, but also has poor taste. In addition, the vinasse also contains a large amount of cellulose, fragrant substances and the like, and at present, the vinasse is mainly used for directly serving as feed, preparing seasonings, culturing edible fungi and the like, and has low additional value. Therefore, how to change the accumulated vinasse like a mountain into valuable through a microorganism channel so as to prepare the feed suitable for feeding animals needs to be solved urgently.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.
The invention also aims to provide a method for preparing yeast culture by using vinasse, which takes vinasse as a substrate, adds mixed bacteria and mixed enzyme liquid, accumulates and ferments to fully decompose macromolecular substances in the vinasse, and then adds bacillus subtilis to increase beneficial bacteria, and simultaneously can promote autolysis of yeast cells by metabolites of the bacillus subtilis, thereby being beneficial to obtaining more yeast cultures.
The invention also aims to provide a yeast culture prepared by the method, which is applied to animal feed, can change the vinasse into valuable, and provides the animal with feed rich in nutrient components so as to improve the immunity of the animal.
To achieve these objects and other advantages in accordance with the purpose of the invention, there is provided a method for preparing a yeast culture using distiller's grains, comprising the steps of:
s1: supplementing a carbon source to the vinasse to obtain a pre-treated vinasse, inoculating a mixed bacterial liquid to the obtained pre-treated vinasse, adding a mixed enzyme liquid to prepare a fermentation base material, and performing fermentation culture for 96-120 h;
s2: inoculating bacillus subtilis after fermentation is finished, continuing fermentation culture for 24 hours, drying, crushing and sieving to obtain a yeast culture;
wherein the mixed bacterial liquid comprises 3 strains of saccharomyces cerevisiae, lactobacillus and aspergillus niger; the mixed enzyme liquid comprises liquefying enzyme, saccharifying enzyme, phytase and protease.
Preferably, the carbon source is corn flour;
and the dried vinasse is mixed with the corn flour according to the weight ratio of 1: 0.1-0.6.
Preferably, after each strain in the mixed bacteria is fermented respectively, a saccharomyces cerevisiae liquid, a lactobacillus liquid and an aspergillus niger liquid are obtained respectively, and then the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:1-3:1-3 to obtain the mixed bacteria liquid;
according to the total weight of the pre-treated distiller's grains, the inoculation amount of the mixed bacterial liquid is 5-20%; the viable count of the mixed bacterial liquid is 1.0 multiplied by 108-1.0×109cfu/mL。
Preferably, the mixed enzyme solution is prepared by mixing a liquefied enzyme solution, a saccharifying enzyme solution, a phytase solution and a protease solution according to a volume ratio of 1: 0.4-0.8: 0.1-0.2: 0.2-1, wherein the enzyme activity of the liquefied enzyme solution, the saccharifying enzyme solution, the phytase solution and the protease solution is 10000U/mL;
adding 1-3% of the mixed enzyme solution based on the total weight of the pre-treated distiller grains.
Preferably, the fermentation conditions in S1 are: spreading the fermentation base material, and turning the fermentation base material once every 5-6h when the temperature is 30-35 ℃, and turning the fermentation base material once every 2-3h when the temperature is increased to be above 40 ℃;
the thickness of the fermentation base material tiling specifically is: when the fermentation temperature is lower than 40 ℃, the tiling thickness is 40-60 cm; when the fermentation temperature rises to 40 ℃ and 40 ℃, the spreading thickness is 20-40 cm.
Preferably, S1 further includes the steps of:
supplementing yeast powder when the fermentation base material is fermented for 72-96 h; the yeast powder is supplemented by 4-10% by weight of the total weight of the pre-treatment distiller's grains.
Preferably, the inoculation amount of the Bacillus subtilis is 1.0 x 10 based on the total weight of the pretreated distiller's grains6cfu/g。
Preferably, the method further comprises the following steps:
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 1000mL of distilled water; fermentation temperature: 27-30 ℃, fermentation conditions: fermenting for 3-5 days at the speed of 200 r/min;
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, 2-5 parts of glucose and 1000mL of distilled water; fermentation temperature: 27-30 ℃, fermentation conditions: fermenting for 3-5 days at the speed of 200 r/min;
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, 5-8 parts of yeast extract, 2-3 parts of diammonium hydrogen citrate, 20-25 parts of glucose, 800.5-1 parts of tween, 5-8 parts of sodium acetate, 2-5 parts of dipotassium hydrogen phosphate, 0.5-1 part of magnesium sulfate, 0.1-0.3 part of manganese sulfate and 1000mL of distilled water, wherein the pH value is 6.2-6.6; fermentation temperature: 35-37 ℃; fermentation conditions are as follows: standing and culturing for 3-5 days;
the fermentation medium of the bacillus subtilis comprises the following components in parts by weight: 15-20 parts of glucose, 30-35 parts of soybean meal, 10-15 parts of starch, 0.1-0.3 part of manganese sulfate, 3-5 parts of dipotassium hydrogen phosphate, 2-3 parts of monopotassium phosphate, 0.05-0.1 part of magnesium sulfate, 2-5 parts of yeast extract, 1-2 parts of ferric chloride, 1-3 parts of calcium carbonate and 1000mL of distilled water, wherein the pH value is 7.0-7.2; fermentation temperature: 35-37 ℃; fermentation conditions are as follows: 200 ℃ and 280r/min, fermenting for 3-5 days.
Preferably, after bacillus subtilis is added into S2, the mixture is spread to be 100cm thick for fermentation, and a material after autolysis is obtained;
drying the autolyzed materials at 85-90 ℃ until the water content is below 10%, crushing and sieving by a 40-mesh sieve to obtain the yeast culture.
The invention also provides application of the yeast culture prepared by the method in animal feed.
The invention at least comprises the following beneficial effects:
the invention takes the vinasse as a matrix, because the vinasse is the residue left after grains are fermented, the content of a carbon source in the residue is low, and if the vinasse is directly used as a fermentation base material, the residue is not beneficial to the survival of strains, the invention firstly supplements a certain carbon source to the vinasse to obtain the pre-treated vinasse, then utilizes various nutrient substances in the pre-treated vinasse to fully degrade by adding a mixed bacterial liquid containing saccharomyces cerevisiae, lactic acid bacteria and aspergillus niger and a mixed bacterial liquid containing liquefying enzyme, saccharifying enzyme, phytase and protease, in particular, because starch, protein, rice husk and the like which participate in the vinasse, the aspergillus niger contains almost complete set of decomposition enzymes aiming at the botanical raw materials, including amylase, lipase, protease, cellulase, xylanase, glucanase, mannase and the like, the plant raw materials can be comprehensively and synergistically decomposed, the yeast, lactobacillus and the like lack the capability of utilizing starch, macromolecular protein, xylan, cellulose, glucan and the like, raw materials containing the components cannot efficiently produce saccharomycetes and lactobacillus, after the raw materials are treated by a series of enzymes generated by aspergillus niger, the starch is decomposed into fermentable glucose, the protein is decomposed into micromolecular peptide or amino acid, the xylan is decomposed into xylose, the glucan, the cellulose and the like are decomposed into glucose monomers, the glucose monomers become nutrition which is easy to absorb and convert by the yeast and the lactobacillus, and after the vinasse raw materials are primarily decomposed by combining with a mixed enzyme agent, the coordination process of the strains is more favorably carried out.
In order to ensure that the fermentation product contains rich micromolecular nutrients and is more suitable for being directly used as animal feed, a certain amount of bacillus subtilis is added after the mixed bacteria are fermented for a period of time, on one hand, beneficial bacteria are added, and on the other hand, metabolic products such as enzymes generated by the bacillus subtilis promote autolysis of yeast cells to generate more yeast cultures. The invention changes the problem that the conventional yeast culture preparation consumes a large amount of carbon sources and nitrogen sources such as molasses, glucose, soybean meal, urea, ammonium bicarbonate and the like, only needs to supplement a small amount of carbon sources, saves resources and greatly reduces the production cost.
In addition, the invention also discloses the application of the prepared yeast culture in animal feed, and the yeast fermentation product prepared by the method mainly contains rich nutrient components, degrades the nutrient substances in the vinasse into small molecular substances suitable for being directly eaten by animals through strains, changes the palatability of the vinasse as the feed, improves the utilization rate of the nutrient substances, and can change a large amount of the vinasse into valuables; the yeast culture of the invention contains a large amount of saccharomyces cerevisiae, and can reach 2.05 multiplied by 109cfu/g, can be directly used as animal feed to feed animals, and improves the immunity of the animals.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Detailed Description
The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
The saccharomyces cerevisiae used in the invention is purchased from Angel Yeast company; the lactobacillus, the aspergillus niger and the bacillus subtilis are all provided by the fermentation engineering room of the university of agriculture in Huazhong.
Example 1
Preparing dried wine lees: drying fresh white spirit vinasse transported from a winery by a dryer at 115 ℃ until the water content is lower than 15% to obtain dried vinasse.
Preparing a saccharomyces cerevisiae bacterial liquid: activating with YPD slant culture medium at 30 deg.C for 20 hr, and fermenting with YPD fermentation culture medium at 30 deg.C and 180r/min for 5 days.
The YPD slant culture medium comprises the following components in parts by weight: 3 parts of yeast extract, 4 parts of peptone, 4 parts of glucose and 1000mL of distilled water;
the 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 1000mL of distilled water.
Preparing an aspergillus niger bacterial liquid: activating with YPD slant culture medium at 30 deg.C for 20 hr, and fermenting with YPD fermentation culture medium at 30 deg.C and 180r/min for 4 days.
The YPD slant culture medium comprises the following components in parts by weight: 3 parts of yeast extract, 5 parts of peptone, 5 parts of glucose and 1000mL of distilled water;
the 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 1000mL of distilled water.
Preparing a lactic acid bacteria liquid: activating with MRS slant culture medium at 35 deg.C for 20 hr, and standing with MRS fermentation culture medium at 37 deg.C for 5 days.
The MRS slant culture medium comprises the following components in parts by weight: 13 parts of peptone, 12 parts of beef extract, 8 parts of yeast extract, 3 parts of diammonium hydrogen citrate, 25 parts of glucose, 801 parts of tween, 8 parts of sodium acetate, 5 parts of dipotassium hydrogen phosphate, 1 part of magnesium sulfate, 0.3 part of manganese sulfate and 1000mL of distilled water, wherein the pH value is 6.6;
the 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, 2 parts of diammonium hydrogen citrate, 20 parts of glucose, 800.5 parts of tween, 6 parts of sodium acetate, 3 parts of dipotassium hydrogen phosphate, 0.5 part of magnesium sulfate, 0.1 part of manganese sulfate and 1000mL of distilled water, wherein the pH value is 6.6.
Preparing a bacillus subtilis liquid: activating with slant culture medium at 35 deg.C for 20 hr, and fermenting with fermentation culture medium at 37 deg.C and 200r/min for 5 days.
Wherein the slant culture medium comprises the following components in parts by weight: 20 parts of glucose, 35 parts of soybean meal, 15 parts of starch, 0.3 part of manganese sulfate, 5 parts of dipotassium hydrogen phosphate, 3 parts of monopotassium phosphate, 0.1 part of magnesium sulfate, 5 parts of yeast extract, 2 parts of ferric chloride, 3 parts of calcium carbonate and 1000mL of distilled water, wherein the pH value is 7.0;
the fermentation medium comprises the following components in parts by weight: 15 parts of glucose, 30 parts of soybean meal, 12 parts of starch, 0.12 part of manganese sulfate, 4 parts of dipotassium hydrogen phosphate, 2 parts of monopotassium phosphate, 0.05 part of magnesium sulfate, 2 parts of yeast extract, 1 part of ferric chloride, 2 parts of calcium carbonate and 1000mL of distilled water, wherein the pH value is 7.0.
Example 2
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
taking 200kg of the distiller's dried grain in example 1, mixing the distiller's dried grain and corn flour according to a weight ratio of 1: 0.2, mixing the dried distillers grains and the corn flour according to the total amount of the dried distillers grains and the corn flour in a material-water ratio of 1:1, uniformly stirring and mixing a mixture of the distiller's dried grains and the corn flour with water, and adjusting the pH value to about 5.0 to obtain pretreated distiller's grains;
according to the total dry basis weight of the pre-treated distiller's grains, 5% of mixed bacteria liquid (in example 1, the saccharomyces cerevisiae bacteria liquid, the lactobacillus bacteria liquid and the aspergillus niger bacteria liquid are mixed according to the volume ratio of 1:1: 1) is added, 1% of mixed enzyme liquid (formed by mixing a liquefied enzyme solution, a saccharifying enzyme solution, a phytase solution and a protease solution according to the volume ratio of 1: 0.4: 0.1: 0.2) is added, a fermentation base material is obtained, 5% of yeast extract powder is supplemented to the fermentation base material after the fermentation for 72h, the stacking thickness of the fermentation initial material is 50cm before the fermentation temperature reaches 40 ℃, and the material is turned once every 5h while the fermentation temperature is kept at 30-35 ℃; when the fermentation temperature reaches 40 ℃ and above 40 ℃, the thickness of the pile is reduced to 10cm, and the material is turned once every 2-3 h.
Inoculating 1.0X 10 after fermentation6cfu/g of Bacillus subtilis, stacking at thickness of 100cm for 24 hr, oven drying at 85 deg.C to water content below 10%, pulverizing, and sieving with 40 mesh sieve to obtain yeast cultureAnd (5) nourishing the food.
The fermentation is finished and before the bacillus subtilis is inoculated, the number of the yeast is measured to be 1.9 multiplied by 10 by adopting a PCA counting method9cfu/g; the total protein amount was 15%.
Example 3
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the embodiment 2 is that: the weight ratio of the distiller's dried grains to the corn flour is 1: 0.3, and the rest steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.93X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 16.7%.
Example 4
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the embodiment 2 is that: the weight ratio of the distiller's dried grains to the corn flour is 1: 0.4, and the rest steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.95X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 17.7%.
Example 5
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the embodiment 2 is that: the weight ratio of the distiller's dried grains to the corn flour is 1: 0.5, and the rest steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.93X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 18.3%.
Example 6
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the example 5 is that: the inoculation amount of the mixed bacteria (the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:2: 2) is 5 percent, and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.96X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 19%.
Example 7
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the example 5 is that: the inoculation amount of the mixed bacteria (the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:2: 2) is 7 percent, and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.98X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 19.5%.
Example 8
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the example 5 is that: the inoculation amount of the mixed bacteria (the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:2: 2) is 10 percent, and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 2.0X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 19.9%.
Example 9
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from example 8 is that: adding 2% of mixed enzyme solution (prepared by mixing the liquefied enzyme solution, the saccharifying enzyme solution, the phytase solution and the protease solution according to the volume ratio of 1: 0.6: 0.2: 1) and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 2.03X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 22%.
Example 10
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from example 8 is that: adding 3% of mixed enzyme solution (prepared by mixing the liquefied enzyme solution, the saccharifying enzyme solution, the phytase solution and the protease solution according to the volume ratio of 1: 0.6: 0.2: 1) and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 2.0X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 25%.
Example 11
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from the example 10 is that: the inoculation amount of the mixed bacteria (the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:3: 3) is 10 percent, 1 percent of mixed enzyme liquid (the liquefied enzyme solution, the saccharifying enzyme solution, the phytase solution and the protease solution are mixed according to the volume ratio of 1: 0.8: 0.2: 1) is added, 8 percent of yeast powder is supplemented, and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 2.05X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 26%.
Example 12
A method of fermenting a yeast culture with distillers grains, comprising the steps of:
the difference from example 11 is that: the inoculation amount of the mixed bacteria (the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to the volume ratio of 1:3: 3) is 20 percent, the yeast powder is supplemented by 10 percent, and the other steps are the same.
The yeast count of the resulting yeast culture was determined to be 1.93X 10 by PCA counting method after the end of fermentation and before inoculation of Bacillus subtilis9cfu/g; the total protein amount was 25.4%.
Example 2-example 12, compared with the prior art, after the mixed strain stacking fermentation is finished, a large amount of yeast is produced, so as to provide a basis for providing a large amount of yeast culture, and then the addition of the bacillus subtilis is used for stacking fermentation, so that the temperature rise is beneficial to the generation of secondary metabolites by the bacillus subtilis, and exogenous substances for promoting the autolysis of yeast cells are provided, so as to be beneficial to obtaining the yeast culture with small molecules which are easy to be absorbed by animals.
Example 13
In a certain pig farm in Shijiazhuang City in Hebei province, the yeast culture prepared in the embodiment 11 is directly used as a feed, and simultaneously 30 pigs with the weight of 50 jin and the same age are divided into a group A, a group B and a group C, wherein the group A is used for feeding the yeast culture prepared in the embodiment 11, the group B is used for feeding the feed purchased in the market, the group C is not used for feeding any feed, the group A and the group B are used for feeding 2 jin of feed every day on the basis of daily ration, and the group C is used for feeding 2 jin on the basis of basic daily ration, and the result shows that: compared with group C, group A shows marketing 20 days earlier, and group B shows marketing 5 days earlier;
before marketing, 3 of group C showed virus infection symptoms, and 2 of the groups were treated to recover to normal state, and 1 of the groups died; group B, 2, after treatment, 1, recovers to normal but the feed intake is obviously reduced, 1, dies, and during the period, 1, still has dysentery and recovers to normal after treatment; group a 10 pigs did not develop any symptoms.
Example 14
In a certain chicken farm in Xinxiang city in Henan province, the yeast culture prepared in example 11 is directly used as a feed, 300 broilers with the same weight and 1-day age number are equally divided into a group A and a group B, wherein the group A is used for feeding the yeast culture prepared in example 11, the group B is used for feeding the feed purchased in the market, the feed with the same amount is fed to the group A and the group B every day after 20 days of observation, and the result shows that the average body weight of the group A of the 20-year-old chickens is 0.3kg higher than that of the group B, no symptoms appear in the group A, and 20 diarrhea phenomena appear in the group B, and the symptoms are recovered after treatment.
While embodiments of the invention have been disclosed above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the generic concept as defined by the claims and their equivalents.
Claims (9)
1. A method of preparing a yeast culture from whole stillage comprising the steps of:
s1: supplementing a carbon source to the vinasse to obtain a pre-treated vinasse, inoculating a mixed bacterial liquid to the obtained pre-treated vinasse, adding a mixed enzyme liquid to prepare a fermentation base material, and performing fermentation culture for 96-120 h;
s2: inoculating bacillus subtilis after fermentation is finished, continuing fermentation culture for 24 hours, drying, crushing and sieving to obtain a yeast culture;
wherein the mixed bacterial liquid comprises 3 strains of saccharomyces cerevisiae, lactobacillus and aspergillus niger; the mixed enzyme liquid comprises a liquefying enzyme, a saccharifying enzyme, a phytase and a protease;
the mixed enzyme solution is prepared from a liquefied enzyme solution, a saccharifying enzyme solution, a phytase solution and a protease solution according to the volume ratio of 1: 0.4-0.8: 0.1-0.2: 0.2-1, wherein the enzyme activity of the liquefied enzyme solution, the saccharifying enzyme solution, the phytase solution and the protease solution is 10000U/mL;
adding 1-3% of the mixed enzyme solution based on the total weight of the pre-treated distiller grains.
2. The method of claim 1, wherein the carbon source is corn meal;
and the dried vinasse is mixed with the corn flour according to the weight ratio of 1: 0.1-0.6.
3. The method for preparing yeast culture by using distiller's grains according to claim 1, wherein after each strain in the mixed bacteria is fermented respectively, a saccharomyces cerevisiae liquid, a lactobacillus liquid and an aspergillus niger liquid are obtained respectively, and then the saccharomyces cerevisiae liquid, the lactobacillus liquid and the aspergillus niger liquid are mixed according to a volume ratio of 1:1-3:1-3 to obtain a mixed bacterial liquid;
according to the total weight of the pre-treated distiller's grains, the inoculation amount of the mixed bacterial liquid is 5-20%; the viable count of the mixed bacterial liquid is 1.0 multiplied by 108-1.0×109cfu/mL。
4. The method of claim 1, wherein the fermentation conditions in S1 are: spreading the fermentation base material, and turning the fermentation base material once every 5-6h when the temperature is 30-35 ℃, and turning the fermentation base material once every 2-3h when the temperature is increased to be above 40 ℃;
the thickness of the fermentation base material tiling specifically is: when the fermentation temperature is lower than 40 ℃, the tiling thickness is 40-60 cm; when the fermentation temperature rises to 40 ℃ or above 40 ℃, the spreading thickness is 20-40 cm.
5. The method for preparing yeast culture using distiller' S grains of claim 1, wherein S1 further comprises the steps of:
supplementing yeast powder when the fermentation base material is fermented for 72-96 h; the yeast powder is supplemented by 4-10% by weight of the total weight of the pre-treatment distiller's grains.
6. The method of preparing a yeast culture using distillers grains according to claim 1, wherein the Bacillus subtilis is inoculated in an amount of 1.0 x 10 based on the total weight of the pretreated distillers grains6cfu/g。
7. The method of preparing a yeast culture using distillers grains according to claim 1, further comprising:
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 1000mL of distilled water; fermentation temperature: 27-30 ℃, fermentation conditions: fermenting for 3-5 days at the speed of 200 r/min;
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, 2-5 parts of glucose and 1000mL of distilled water; fermentation temperature: 27-30 ℃, fermentation conditions: fermenting for 3-5 days at the speed of 200 r/min;
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, 5-8 parts of yeast extract, 2-3 parts of diammonium hydrogen citrate, 20-25 parts of glucose, 800.5-1 parts of tween, 5-8 parts of sodium acetate, 2-5 parts of dipotassium hydrogen phosphate, 0.5-1 part of magnesium sulfate, 0.1-0.3 part of manganese sulfate and 1000mL of distilled water, wherein the pH value is 6.2-6.6; fermentation temperature: 35-37 ℃; fermentation conditions are as follows: standing and culturing for 3-5 days;
the fermentation medium of the bacillus subtilis comprises the following components in parts by weight: 15-20 parts of glucose, 30-35 parts of soybean meal, 10-15 parts of starch, 0.1-0.3 part of manganese sulfate, 3-5 parts of dipotassium hydrogen phosphate, 2-3 parts of monopotassium phosphate, 0.05-0.1 part of magnesium sulfate, 2-5 parts of yeast extract, 1-2 parts of ferric chloride, 1-3 parts of calcium carbonate and 1000mL of distilled water, wherein the pH value is 7.0-7.2; fermentation temperature: 35-37 ℃; fermentation conditions are as follows: 200 ℃ and 280r/min, fermenting for 3-5 days.
8. The method of claim 1, wherein the yeast culture is prepared from distillers' grains, wherein the S2 is added with Bacillus subtilis and then fermented by spreading to a thickness of 100cm to obtain autolyzed material;
drying the autolyzed materials at 85-90 ℃ until the water content is below 10%, crushing and sieving by a 40-mesh sieve to obtain the yeast culture.
9. Use of a yeast culture prepared by the method of any one of claims 1 to 8 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 | A kind of fermentation method of yeast culture using Maotai-flavor liquor grains as raw material |
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| CN113621674B (en) * | 2021-08-27 | 2022-06-21 | 泸州老窖股份有限公司 | Method for producing L-lactic acid by using liquor distiller grains |
| CN113892556B (en) * | 2021-09-29 | 2024-07-09 | 四川大学青岛研究院 | Biological protein feed prepared from waste distillers' grains of Jimo old wine and preparation method thereof |
| CN113999781A (en) * | 2021-09-29 | 2022-02-01 | 宁夏未来生物科技有限公司 | Environment-friendly yeast production process |
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