CN113430131B - Koji block for fermented feed and preparation method thereof - Google Patents

Koji block for fermented feed and preparation method thereof Download PDF

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CN113430131B
CN113430131B CN202110681473.2A CN202110681473A CN113430131B CN 113430131 B CN113430131 B CN 113430131B CN 202110681473 A CN202110681473 A CN 202110681473A CN 113430131 B CN113430131 B CN 113430131B
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CN113430131A (en
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岳支华
陈红坤
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Beijing Ailan Biotechnology Co ltd
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Abstract

The application relates to the field of microbial fermentation, and particularly discloses a yeast block for fermented feed and a preparation method thereof; the yeast block is prepared from the following raw materials in parts by weight: 55-68 parts of yeast block material, 8-16 parts of mixed bacterial powder and 36-54 parts of water; the mixed bacteria powder consists of mould, bacillus subtilis, saccharomycetes, bacillus coagulans and clostridium butyricum; the preparation method comprises the following steps: weighing clostridium butyricum, yeast block materials with the weight of 1/8-1/4 and water with the total amount of 1/10-1/6, mixing and stirring to prepare granular molding materials; spraying a film forming solution on the surface of the molding material to prepare a primary mixed material; weighing mould, bacillus subtilis, saccharomycete and bacillus coagulans, mixing and stirring the mould, the bacillus subtilis, the saccharomycete and the bacillus coagulans with the rest of the koji material and the rest of the water, then adding the initial mixture, continuously stirring, and preparing a finished koji block through compression molding and after-fermentation; has the advantages of multi-bacteria symbiosis, enzyme system complementation, reduction of the production cost of mixed fermentation strains prepared by enterprises and the like.

Description

Koji block for fermented feed and preparation method thereof
Technical Field
The application relates to the field of microbial fermentation, in particular to a yeast block for fermenting feed and a preparation method thereof.
Background
The anti-nutritional factors such as soybean anti-nutritional factors, non-starch polysaccharides, phytic acid, cellulose, hemicellulose, pectin substances and the like contained in the feed raw materials are difficult to be directly digested and absorbed by animals, can increase the burden of intestinal tracts and even easily cause intestinal diseases if not treated; to achieve a better replacement effect, anti-nutritional factors in the feed need to be eliminated, and macromolecular substances in the feed need to be decomposed.
The common method is that the raw materials of the feed are fermented by microorganisms and then fed to animals, and the fermented feed not only contains a plurality of beneficial microorganisms, but also can decompose antinutritional factors in the feed; protease, saccharifying enzyme, cellulase, etc. produced by microorganism metabolism can decompose part of protein, cellulose, starch polysaccharide, etc. into small peptide, oligosaccharide, etc. to promote digestion and absorption of animal intestinal tract.
In the preparation process of the fermented feed in the related technology, a pure fermentation mode is usually adopted for production; the fermented feed is prepared by mixing a raw material substance with a fermented product and other raw materials, and the pure fermentation uses a single microorganism, so that the enzyme systems participating in biochemical reaction are few, the metabolites are few, the nutrient substances contained in the other raw materials in the fermented raw materials cannot be effectively decomposed, and the nutrient substances in the other raw materials cannot be easily absorbed and digested by animals after the animals eat the fermented feed; therefore, in order to decompose all the nutrients in the raw materials of fermented feed, the fermented feed is generally prepared by mixed fermentation.
The mixed fermentation strain is prepared by adopting single strain liquid to produce, then carrying out a series of post-treatments such as concentration and drying, and the like, and mixing a plurality of prepared single strains to prepare a mixed fermentation strain; a plurality of fermentation tanks are needed to be used simultaneously in the production process of the mixed fermentation strain, the fermentation tanks are utilized to respectively correspond to a plurality of strains for fermentation, and the fermentation periods are different due to different fermentation conditions of different strains; therefore, the process of preparing the mixed fermentation strain is complicated and high in cost, and the production cost of enterprises is increased.
Disclosure of Invention
In order to reduce the production cost of preparing mixed fermentation strains by enterprises, the application provides a yeast block for fermented feed and a preparation method thereof.
In a first aspect, the present application provides a koji block for fermented feed, which adopts the following technical scheme:
a koji block for fermented feed, which is prepared from the following raw materials in parts by weight: 55-68 parts of yeast block material, 8-16 parts of mixed bacterial powder and 36-54 parts of water; the mixed bacteria powder consists of mould, bacillus subtilis, saccharomycete, bacillus coagulans and clostridium butyricum in the weight ratio of 0.4-1.6 to 1:0.4-1.6 to 0.5-1.5.
Through adopting above-mentioned technical scheme, utilize bent piece material, mixed fungus powder, water mixture preparation bent piece, at the fermentation process, the different positions department simultaneous growth that multiple fungus in the mixed fungus powder can be on bent piece breeds, need not a plurality of fermentation tanks and cultivates different fungus to reduce the manufacturing cost of enterprise preparation mixed fermentation bacterial.
The surface of the koji block is contacted with oxygen, and aerobic bacteria such as mould, bacillus subtilis and the like in the mixed bacteria powder can quickly grow and propagate on the surface of the koji block; the inside of the koji block is basically free of oxygen, and anaerobic bacteria clostridium butyricum and bacillus coagulans in the mixed strains quickly grow and propagate in the koji block; the middle part of the koji block close to the surface of the koji block is contacted with oxygen, the middle part of the koji block close to the inner part of the koji block basically has no oxygen, and facultative anaerobes such as saccharomycetes, bacillus coagulans and the like in the mixed bacteria powder rapidly grow and propagate in the middle part of the koji block; namely, a plurality of bacteria in the mixed bacteria powder can simultaneously grow and reproduce at different positions on the koji block.
Facultative anaerobes such as saccharomycetes and the like can grow and propagate rapidly under aerobic conditions, oxygen in the middle of a koji block and oxygen in the koji block are consumed rapidly in the growth and propagation processes, growth and propagation of anaerobic bacteria clostridium butyricum and bacillus coagulans are promoted while growth and propagation of the anaerobic bacteria are promoted.
Preferably, the mold consists of aspergillus niger and aspergillus oryzae; the yeast comprises Saccharomyces cerevisiae and Candida utilis.
By adopting the technical scheme, aspergillus niger and aspergillus oryzae are matched, so that the fermented feed is rich in amylase, protease, pectinase, cellulase and other substances, and starch, fiber and protein are decomposed into small molecular substances; more nutrition is provided for the growth of saccharomyces cerevisiae and candida utilis, and more metabolites are produced; the digestion and absorption rate of the fermented feed is improved, and the nutritive value of the fermented feed is improved.
Preferably, the koji block material is prepared from the following raw materials in parts by weight: 15-25 parts of corn flour, 14-24 parts of soybean meal, 46-55 parts of wheat flour, 5-12 parts of molasses and 0.6-1.8 parts of mineral substances.
By adopting the technical scheme, the corn flour and the wheat flour are matched to mainly provide starch for the mixed bacterial powder, the corn flour and the soybean meal are matched to mainly provide protein for the mixed bacterial powder, the molasses provides saccharides such as glucose and the like, and the mineral substance provides mineral elements; the growth and reproduction of thalli in the mixed bacterial powder are promoted by providing sufficient energy substances.
Preferably, the mineral substance consists of sodium chloride and dipotassium hydrogen phosphate in a weight ratio of 1: 0.6-1.5.
By adopting the technical scheme, sodium chloride and dipotassium phosphate are matched to provide sodium ions for the yeast blocks, and the sodium ions can maintain the osmotic pressure of cells, so that the thalli in the mixed bacterial powder can be ensured to grow and reproduce quickly; and in the process of growth and reproduction of thalli in the mixed bacterial powder, a plurality of acidic metabolites can be generated, and the alkaline of dipotassium hydrogen phosphate is utilized to neutralize acidic substances, so that the thalli in the mixed bacterial powder can be ensured to grow and reproduce quickly.
In a second aspect, the present application provides a method for preparing a koji block for fermented feed, which adopts the following technical scheme: a preparation method of a yeast block for fermented feed comprises the following steps:
s1, weighing clostridium butyricum, yeast block materials with the weight of 1/8-1/4 and water with the total amount of 1/10-1/6, mixing and stirring to prepare granular molding materials; spraying a film forming solution on the surface of the molding material to prepare a primary mixed material;
s2, weighing mould, bacillus subtilis, saccharomycete and bacillus coagulans, mixing and stirring the mould, the bacillus subtilis, the saccharomycete and the bacillus coagulans with the rest of the yeast cake materials and the rest of the water, then adding the initial mixture prepared in the S1, continuously stirring, and performing compression molding and after-fermentation to obtain the finished product yeast cake.
By adopting the technical scheme, the clostridium butyricum, partial koji block materials and part of water are mixed to preliminarily prepare the molding material, the surface of the molding material is sprayed with the film forming solution, and the growth and the propagation of the clostridium butyricum in the koji block are further promoted by utilizing the isolation effect of the film forming solution on oxygen.
The granular molding material is mixed with other materials, and the moist viscosity effect of the film forming solution is utilized to ensure that other bacteria powder and the koji block material are more tightly bonded on the surface of the molding material, so that the oxygen amount in the pores of the koji block structure is reduced, and the growth and the propagation of clostridium butyricum are further ensured.
Aspergillus niger and Aspergillus oryzae can be used for producing amylase, protease, cellulase, pectinase and other substances, and the digestibility of the fermented feed is improved; the saccharomyces cerevisiae and the candida utilis can utilize raw materials such as starch, protein, molasses and the like, and metabolites of the saccharomyces cerevisiae and the candida utilis can effectively stimulate the feed intake of animals; the bacillus subtilis, the bacillus coagulans and the clostridium butyricum are matched, so that the growth of beneficial flora in intestinal tracts of animals can be promoted, and the growth of harmful bacteria can be inhibited; the finished product of the fermented feed has higher production value.
Preferably, the film forming solution in S1 is prepared by the following method:
weighing 6-14 parts of wheat gluten protein, 2-5 parts of glycerol, 0.2-0.8 part of tween 80, 0.2-1 part of guar gum and 90-110 parts of ethanol, stirring and mixing, and performing vacuum degassing to prepare a film forming solution.
By adopting the technical scheme, the wheat gluten protein, the Tween 80, the guar gum and the glycerol are matched, and the network cross-linking structure formed by the protein, the Tween 80 and the guar gum is utilized to ensure that the film forming solution has good oxygen barrier rate after being dried and formed into a film, so that the better oxygen barrier effect is achieved, oxygen can be better prevented from contacting with the clostridium butyricum, and the growth and the propagation of the clostridium butyricum are further promoted.
The metabolite butyric acid of the clostridium butyricum is a main nutrient substance for regeneration and repair of intestinal epithelial tissue cells, the clostridium butyricum can promote the growth of intestinal beneficial bacteria and inhibit the growth of harmful bacteria, and the clostridium butyricum has higher reproductive capacity and can better promote animals to absorb the nutrient substances in the fermented feed.
Preferably, the total amount of water in S1 is 1/10-1/6, and the rest is tap water.
By adopting the technical scheme, the starch aqueous solution and the yeast block material are matched, and the yeast block material-taking powder in the molding material is tightly bonded by utilizing the better bonding effect of the starch aqueous solution, so that the oxygen content in the molding material is reduced, and the growth and the propagation of clostridium butyricum are promoted; and the starch is matched with corn flour and wheat flour in the koji block material, so that nutrient substances can be further provided for the growth and the propagation of the clostridium butyricum, and the growth and the propagation of the clostridium butyricum are further promoted.
Preferably, modified diatomite and lactose are added into the S1, and the weight ratio of the lactose to the diatomite to the yeast block material in the S1 is 0.02-0.08:0.02-0.15: 1.
By adopting the technical scheme, the modified diatomite, the lactose and the molasses are matched, and the porous structure of the diatomite can be used for further providing space position storage for the growth and the propagation of the clostridium butyricum, so that the growth and the propagation of the clostridium butyricum are promoted; the clostridium butyricum ferments sugar to produce acid, and the metabolite butyric acid is a main nutrient substance for regeneration and repair of intestinal epithelial cells, so that the content of the metabolite is increased, and the nutritional value of the fermented feed is improved.
Preferably, the modified diatomite is prepared by the following method:
weighing kieselguhr, placing the kieselguhr into a magnesium sulfate aqueous solution with the mass fraction of 2-8%, wherein the mass ratio of the kieselguhr to the magnesium sulfate aqueous solution is 1:0.8-1.5, stirring, and taking out the kieselguhr adsorbed with magnesium sulfate to prepare the modified kieselguhr.
By adopting the technical scheme, the magnesium sulfate aqueous solution is absorbed by the diatomite by utilizing the porous adsorption characteristic of the diatomite, so that the influence on the growth and the propagation of clostridium butyricum caused by the absorption of oxygen in the external environment by the porous structure of the diatomite is avoided.
The modified diatomite is matched with the lactose, and the growth and the propagation of clostridium butyricum are further promoted by utilizing nutrient substances in the magnesium sulfate and the lactose; magnesium sulfate in the modified diatomite is consumed in the growth and propagation processes of the clostridium butyricum, and after the magnesium sulfate aqueous solution is separated from the pores of the modified diatomite, the pores of the modified diatomite can provide a growth space for the growth and propagation of the clostridium butyricum, so that the growth and propagation of the clostridium butyricum are promoted.
Preferably, the post-fermentation temperature in S2 is 30-35 deg.C, humidity is 80-90%, and fermentation time is 10-20 days.
By adopting the technical scheme, a large amount of multiple bacteria in the composite bacteria powder can grow and propagate by limiting the temperature, humidity and time of after-fermentation, so that the quantity of bacteria on the koji block and the enzyme yield are improved.
In summary, the present application has the following beneficial effects:
1. the yeast block is prepared by mixing the yeast block material, the mixed bacteria powder and water, and in the fermentation process, various bacteria in the mixed bacteria powder can grow and propagate at different positions on the yeast block at the same time without culturing different bacteria by a plurality of fermentation tanks.
2. The clostridium butyricum, partial koji block materials and part of water are mixed to preliminarily prepare molding materials, film forming solution is sprayed on the surface of the molding materials, and the growth and the propagation of the clostridium butyricum in the koji block are further promoted by utilizing the isolation effect of the film forming solution on oxygen.
3. Magnesium sulfate in the modified diatomite is consumed in the growth and propagation processes of the clostridium butyricum, and after the magnesium sulfate aqueous solution is separated from the pores of the modified diatomite, the pores of the modified diatomite can provide a growth space for the growth and propagation of the clostridium butyricum, so that the growth and propagation of the clostridium butyricum are promoted.
Detailed Description
The present application will be described in further detail with reference to examples.
Preparation example of film Forming solution
Wheat gluten protein in the following raw materials was purchased from Tianjin, Rizhuanghe chemical technology, Inc.; the protein content was 70.73%; tween 80 was purchased from north Heibei Runshou Biotech limited; guar gum is purchased from Zhengzhou field chemical products, Inc.; other raw materials and equipment are all sold in the market.
Preparation example 1: the film forming solution is prepared by the following method:
weighing 10.8g of wheat gluten protein, 3g of glycerol, 0.5g of Tween 80, 0.5g of guar gum and 102g of ethanol with the mass fraction of 57%, stirring for 5min at the rotating speed of 500r/min, adjusting the pH of the solution to 10 by using sodium hydroxide while stirring, stirring for 30min in a constant-temperature water bath at the temperature of 80 ℃, then carrying out ultrasonic treatment for 15min, and degassing for 10min under the vacuum degree of 0.09MPa to prepare a film forming solution.
Preparation example 2: the difference between the preparation example and the preparation example 1 is that:
weighing 6g of wheat gluten protein, 2g of glycerol, 0.2g of Tween 80, 0.2g of guar gum and 90g of ethanol with the mass fraction of 57%.
Preparation example 3: the difference between the preparation example and the preparation example 1 is that:
weighing 14g of wheat gluten protein, 5g of glycerol, 0.8g of Tween 80, 1g of guar gum and 110g of ethanol with the mass fraction of 57%.
Preparation example of modified diatomaceous earth
The diatomite in the following raw materials is purchased from Kaiyao mineral product processing factory in Lingshu county; magnesium sulfate was purchased from feed grade magnesium sulfate heptahydrate, produced by liangyong fengtai biotechnology limited; other raw materials and equipment are all sold in the market.
Preparation example 4: the modified diatomite is prepared by the following method:
weighing 1kg of diatomite, placing the diatomite into 1kg of magnesium sulfate aqueous solution with the mass fraction of 5%, stirring for 3min at the rotating speed of 800r/min, and taking out the diatomite adsorbed with the magnesium sulfate after stirring to obtain the modified diatomite.
Preparation example 5: the modified diatomite is prepared by the following method:
weighing 1kg of diatomite, placing the diatomite into 0.8kg of magnesium sulfate aqueous solution with mass fraction of 8%, stirring for 3min at the rotating speed of 800r/min, and taking out the diatomite adsorbed with the magnesium sulfate after stirring to obtain the modified diatomite.
Preparation example 6: the modified diatomite is prepared by the following method:
weighing 1kg of diatomite, placing the diatomite into 1.5kg of magnesium sulfate aqueous solution with the mass fraction of 2%, stirring for 3min at the rotating speed of 800r/min, and taking out the diatomite adsorbed with the magnesium sulfate after stirring to obtain the modified diatomite.
Preparation example of koji Material
The following raw materials are all commercially available.
Preparation example 7: the yeast block material is prepared by the following method:
weighing 20kg of corn flour, 20kg of soybean meal, 50kg of wheat flour, 9kg of molasses and 1kg of mineral substances, and stirring at the rotating speed of 1000r/min for 2min to obtain a koji material; the mineral substance comprises sodium chloride and dipotassium hydrogen phosphate in a weight ratio of 1: 1; the crushing fineness of the corn flour, the soybean meal and the wheat flour is 2 mm.
Preparation example 8: the difference between this preparation and preparation 7 is that:
weighing 15kg of corn flour, 14kg of soybean meal, 55kg of wheat flour, 5kg of molasses and 0.6kg of mineral substances; the mineral comprises sodium chloride and dipotassium hydrogen phosphate at a weight ratio of 1: 0.6.
Preparation example 9: the difference between this preparation and preparation 7 is that:
weighing 25kg of corn flour, 24kg of soybean meal, 46kg of wheat flour, 12kg of molasses and 1.8kg of mineral substances; the mineral comprises sodium chloride and dipotassium hydrogen phosphate at a weight ratio of 1: 1.5.
Examples
The following raw materials, Aspergillus niger and Aspergillus oryzae, are purchased from Shandong Kangyuan Biotech limited company, and the fungus content in Aspergillus niger powder and Aspergillus oryzae powder is not less than 2 x 108cfu/g; bacillus subtilis, Saccharomyces cerevisiae, Candida utilis, Bacillus coagulans and Clostridium butyricum are all purchased from Qingdao root biotechnology group Limited company, and the bacterium content in the bacillus subtilis powder is not less than 1 x 1010cfu/g, content of Saccharomyces cerevisiae powder and Candida utilis powderThe quantity is more than or equal to 5 x 109cfu/g, the bacterial content of the bacillus coagulans powder and the clostridium butyricum powder is more than or equal to 1 x 109cfu/g; other raw materials and equipment are all sold in the market.
Example 1: a koji for fermented feed:
weighing 64kg of yeast block material, 12kg of mixed bacterial powder and 48kg of water; the mixed bacterial powder consists of aspergillus niger, aspergillus oryzae, bacillus subtilis, saccharomyces cerevisiae, candida utilis, bacillus coagulans and clostridium butyricum in a weight ratio of 0.5:0.5:1:0.5:0.5:1: 1; the koji material was the koji material prepared in preparation example 7;
the preparation method comprises the following steps:
s1, weighing Aspergillus niger, Aspergillus oryzae, Bacillus subtilis, Saccharomyces cerevisiae, Candida utilis, Bacillus coagulans and Clostridium butyricum, mixing to obtain mixed powder,
s2, stirring the mixed bacterial powder prepared in the step S1, a yeast block material and water at the rotating speed of 800r/min for 15min, then pressing and molding, wherein the size is 20cm multiplied by 15cm multiplied by 7cm, and then performing after-fermentation for 15 days under the conditions that the temperature is 32 ℃ and the humidity is 85% to obtain the finished yeast block.
Example 2: the present embodiment is different from embodiment 1 in that:
55kg of yeast block material, 8kg of mixed bacterial powder and 36kg of water; the mixed bacterial powder consists of aspergillus niger, aspergillus oryzae, bacillus subtilis, saccharomyces cerevisiae, candida utilis, bacillus coagulans and clostridium butyricum in a weight ratio of 0.2:0.8:1:0.2:0.8:0.5: 1.5.
Example 3: the present embodiment is different from embodiment 1 in that:
68kg of yeast block material, 16kg of mixed bacterial powder and 54kg of water; the mixed bacterial powder consists of aspergillus niger, aspergillus oryzae, bacillus subtilis, saccharomyces cerevisiae, candida utilis, bacillus coagulans and clostridium butyricum in a weight ratio of 0.8:0.2:1:0.8:0.2:1.5: 0.5.
Example 4: the present embodiment is different from embodiment 1 in that:
selecting the koji material prepared in preparation example 8; the post-fermentation temperature is 30 deg.C, humidity is 80%, and the time is 20 days.
Example 5: the present embodiment is different from embodiment 1 in that:
the koji material prepared in preparation example 9 was selected as the koji material; the post-fermentation temperature is 35 deg.C, humidity is 90%, and the time is 10 days.
Example 6: the present embodiment is different from embodiment 1 in that: a yeast block for fermented feed is prepared by the following method:
s1, weighing 2.4kg of clostridium butyricum, 12.8kg of yeast block material and 6kg of starch water solution, mixing, stirring for 3min at the rotating speed of 500r/min, and granulating to obtain a granular molding material, wherein the diameter of the molding material is 1 cm; spraying the film forming solution prepared in preparation example 1 on the surface of the molding material, wherein the weight ratio of the molding material to the film forming solution is 1:0.2, and preparing a primary mixed material;
s2, weighing 1.2kg of Aspergillus niger, 1.2kg of Aspergillus oryzae, 2.4kg of Bacillus subtilis, 1.2kg of Saccharomyces cerevisiae, 1.2kg of Candida utilis, 2.4kg of Bacillus coagulans, 51.2kg of yeast block material and 42kg of water, mixing at the rotating speed of 800r/min for 10min, adding the initial mixture prepared in S1, continuing stirring for 5min, then performing compression molding, wherein the size is 20cm multiplied by 15cm multiplied by 7cm, and performing after-fermentation for 10 days under the conditions that the temperature is 30 ℃ and the humidity is 80% to obtain the finished yeast block.
Example 7: this embodiment is different from embodiment 6 in that:
s1, weighing 2.4kg of clostridium butyricum, 8kg of yeast block material and 4.8kg of starch water solution and mixing;
s2, weighing 1.2kg of Aspergillus niger, 1.2kg of Aspergillus oryzae, 2.4kg of Bacillus subtilis, 1.2kg of Saccharomyces cerevisiae, 1.2kg of Candida utilis, 2.4kg of Bacillus coagulans, 56kg of yeast cake material and 43.2kg of water to mix.
Example 8: this embodiment is different from embodiment 6 in that:
s1, weighing 2.4kg of clostridium butyricum, 16kg of yeast block materials and 8kg of starch water solution, and mixing;
s2, weighing 1.2kg of Aspergillus niger, 1.2kg of Aspergillus oryzae, 2.4kg of Bacillus subtilis, 1.2kg of Saccharomyces cerevisiae, 1.2kg of Candida utilis, 2.4kg of Bacillus coagulans, 40kg of yeast cake material and 40kg of water, and mixing.
Example 9: this embodiment is different from embodiment 6 in that:
the film-forming solution prepared in preparation example 2 was used as the film-forming solution.
Example 10: this embodiment is different from embodiment 6 in that:
the film-forming solution prepared in preparation example 3 was used as the film-forming solution.
Example 11: this embodiment is different from embodiment 6 in that:
s1, weighing 2.4kg of clostridium butyricum, 12.8kg of yeast block material, 6kg of starch water solution, 0.64kg of lactose and 1.28kg of modified diatomite prepared in preparation example 4, mixing, and stirring at the rotating speed of 500r/min for 3 min.
Example 12: this embodiment is different from embodiment 6 in that:
s1, weighing 2.4kg of clostridium butyricum, 12.8kg of yeast block material, 6kg of starch water solution, 0.256kg of lactose and 0.256kg of modified diatomite prepared in the preparation example 4, mixing, and stirring at the rotating speed of 500r/min for 3 min.
Example 13: this embodiment is different from embodiment 6 in that:
s1, weighing 2.4kg of clostridium butyricum, 12.8kg of yeast block material, 6kg of starch water solution, 1.024kg of lactose and 1.92kg of modified diatomite prepared in the preparation example 4, mixing, and stirring at the rotating speed of 500r/min for 3 min.
Example 14: this embodiment is different from embodiment 11 in that:
modified diatomaceous earth the modified diatomaceous earth prepared in preparation example 5 was used.
Example 15: this embodiment is different from embodiment 11 in that:
modified diatomaceous earth the modified diatomaceous earth prepared in preparation example 6 was used.
Example 16: this embodiment is different from embodiment 6 in that:
the raw material of the film forming solution is used for replacing Tween 80 by guar gum with the same mass.
Example 17: this embodiment is different from embodiment 6 in that:
the water mixed with clostridium butyricum in S1 is normal tap water.
Example 18: this embodiment is different from embodiment 11 in that:
the modified diatomite is replaced by lactose with the same mass in the raw materials.
Example 19: this embodiment is different from embodiment 11 in that:
the diatomite is commercially available diatomite produced by Kaiyao mineral product processing factories in Lingshu county.
Example 20: the present embodiment is different from embodiment 1 in that:
the mould is Aspergillus niger and the yeast is Saccharomyces cerevisiae.
Comparative example
Comparative example 1: this comparative example differs from example 1 in that: the mould is replaced by the same mass of bacillus subtilis in the raw materials.
Comparative example 2: this comparative example is different from example 1 in that: the raw materials are used for replacing bacillus coagulans by yeast with the same mass.
Comparative example 3: this comparative example differs from example 1 in that: the clostridium butyricum is replaced by bacillus subtilis with the same mass in the raw materials.
Performance test
1. Detection of microbial quantity in koji
Preparing koji blocks by respectively adopting the preparation methods of examples 1-20, respectively detecting the quantity of melanomyces and aspergillus oryzae in the koji blocks by adopting a method for detecting the counts of moulds and yeasts by GB478915-2010 food microbiology and a method for detecting the morphological identification of common toxin-producing moulds by GB4789.16-2016 food safety national standard food microbiology, respectively detecting the quantity of saccharomyces cerevisiae by adopting the standard of active dry yeast (saccharomyces cerevisiae) for feed of a GBT22547-2008 feed additive, and detecting the quantity of candida utilis by adopting the standard of NY/T1969-; detecting the quantity of the bacillus subtilis in the koji block by adopting the standard for detecting the bacillus subtilis in the GB/T26428-2010 feeding microbial preparation; detecting the number of bacillus coagulans in the koji block by adopting a method for detecting the bacillus coagulans in the DB21/T3278-2020 feeding microbial preparation; and detecting the number of clostridium butyricum in the koji block by using a DB41/T1728-2018 feed additive clostridium butyricum determination microbiological method.
TABLE 1 detection of microbial numbers
Figure BDA0003122781890000091
TABLE 2 detection of microbial numbers
Figure BDA0003122781890000092
Figure BDA0003122781890000101
As can be seen by combining the examples 1-3 and the tables 1 and 2, the yeast blocks prepared by the method can produce various strains at the same time, the number of the strains is high, and the production cost of preparing mixed fermentation strains by enterprises can be reduced by preparing the fermented feed by the yeast blocks.
Example 1 in combination with examples 4-5 and tables 1 and 2, it can be seen that the growth and propagation of each thallus on the koji blocks are easily affected due to different koji block raw materials and different post-fermentation conditions.
Example 1, in combination with examples 6 to 8 and tables 1 and 2, it can be seen that the number of clostridium butyricum on the koji blocks prepared in examples 6 to 8 is increased, which indicates that oxygen absorbed by the koji blocks in the preparation process can be blocked by the blocking effect of the film forming solution, and the growth and reproduction of clostridium butyricum can be promoted under the condition of better oxygen blocking.
Example 6 in combination with examples 9-10 and tables 1 and 2, it can be seen that the amount of clostridium butyricum of examples 9-10 is less than that of example 6, indicating that the oxygen barrier property of the film forming solution easily affects the growth and reproduction of clostridium butyricum on the koji.
Example 6 in combination with examples 11-13 and tables 1 and 2, it can be seen that the addition of modified diatomaceous earth and lactose to the starting materials of examples 11-13, compared to example 6, resulted in the production of a greater number of Clostridium butyricum in the koji blocks of examples 11-13 than in example 6; the modified diatomite, the lactose and the molasses are matched, and the porous structure of the diatomite can be used for further providing space position storage for the growth and the propagation of the clostridium butyricum, so that the growth and the propagation of the clostridium butyricum are promoted.
Example 11 in combination with examples 14-15 and tables 1 and 2, it can be seen that the amount of clostridium butyricum in koji blocks prepared in examples 14-15 is lower than that in example 11, which shows that the loading of magnesium sulfate by modified diatomite has influence on the growth and reproduction of clostridium butyricum in koji blocks.
Example 6 in combination with examples 16-17 and tables 1 and 2, it can be seen that the amount of clostridium butyricum in the koji prepared in example 16 is lower than that in example 6 compared with example 6 when tween 80 is replaced by guar gum of the same quality in the raw material of the film forming solution in example 16; the matching of the guar gum and the Tween 80 is proved, so that the film forming solution has good oxygen blocking effect after being cured into a film, and the growth and the propagation of the clostridium butyricum are further promoted under the anaerobic condition.
Example 17 in the preparation of the koji, water mixed with clostridium butyricum in S1 was normal tap water, and the number of clostridium butyricum in the koji prepared in example 17 was lower than that in example 6 compared with example 6; the cooperation of the starch aqueous solution and the yeast block materials is demonstrated, the yeast block taking powder in the molding material is tightly bonded by utilizing the better bonding effect of the starch aqueous solution, the oxygen content in the molding material is reduced, the growth and the propagation of clostridium butyricum are promoted, and meanwhile, nutrient substances can be provided, and the growth and the propagation of clostridium butyricum are further promoted.
Example 11 in combination with examples 18-19 and tables 1 and 2, it can be seen that the modified diatomaceous earth was replaced by an equivalent mass of lactose in the feed of example 18, and the number of clostridium butyricum in koji prepared in example 18 was lower than that of example 11 compared to example 11; the matching of the lactose and the modified diatomite can promote the growth and the propagation of the clostridium butyricum on the koji.
Example 19 the diatomaceous earth in the raw material is commercially available diatomaceous earth, and compared with example 11, the number of clostridium butyricum on koji prepared in example 19 is lower than that of example 11; the diatomite pore is loaded with magnesium sulfate, the magnesium sulfate in the pore is gradually absorbed in the growth and reproduction process of the clostridium butyricum, and the diatomite pore can provide space position storage for newly grown and reproduced clostridium butyricum, so that the growth and reproduction of the clostridium butyricum are promoted.
Example 1 in combination with example 20 and tables 1 and 2, it can be seen that the mold in the raw material of example 20 is aspergillus niger and the yeast is saccharomyces cerevisiae, and compared with example 1, the number of aspergillus niger on the koji block prepared in example 20 is less than the sum of the numbers of aspergillus niger and aspergillus oryzae prepared in example 1; the aspergillus niger and the aspergillus oryzae are matched to promote the absorption and utilization of the thalli to the raw materials and promote the growth and the propagation of the thalli; the number of saccharomyces cerevisiae on the koji prepared in example 20 is less than the sum of saccharomyces cerevisiae and candida utilis of example 1; the combination of the saccharomyces cerevisiae and the candida utilis is proved to be capable of promoting the growth and the propagation of thalli.
Detection of oxygen barrier property of film forming solution
The film-forming solutions were prepared by the preparation methods of preparation examples 1 to 3 and example 16, respectively, and 10mL of the film-forming solution was measured and poured in 100cm2The sample is uniformly spread on the glass plate, dried for 2.5 hours at the temperature of 60 ℃, taken out and stored in a hygrostat (a saturated solution of magnesium nitrate) with the relative humidity of 54 percent for 2 days to prepare the sample to be tested.
Weighing 5.0g of soybean oil, placing the soybean oil in a beaker, covering a sample on the beaker, sealing the beaker by vaseline, taking the soybean oil which is not covered with any substance as a reference, placing the beaker in a blast drying oven at 60 ℃, storing the beaker for 7 days, removing a film, weighing 2.00g of an oil sample, and detecting the peroxide value of the oil sample.
Weighing 2.00g of soybean oil according to GB/T5009.37-2003, placing the soybean oil into a 250mL iodine measuring flask, adding 30mL of chloroform/glacial acetic acid mixed solution, shaking to completely dissolve a sample, adding 1mL of saturated potassium iodide solution, tightly covering the flask cover, slightly shaking for 30s, then placing the flask in the dark for 3min, taking out the flask, adding 100mL of water, shaking uniformly, immediately titrating the mixture to light yellow by using a sodium thiosulfate standard solution, adding 1mL of starch indicator, continuing to titrate until blue disappears as an end point, performing a blank test, and recording data to reflect the oxygen blocking condition of the sample.
TABLE 3 oxygen-blocking condition testing table
Item Oxygen transmission rate/meq/kg
Preparation example 1 11.02±0.08
Preparation example 2 11.45±0.17
Preparation example 3 10.98±0.84
Example 16 15.42±0.68
Preparation examples 1 to 3 in combination with example 16 and table 3, it can be seen that guar gum and tween 80 are matched to make the film forming solution have good oxygen barrier property after film forming.
Enzyme activity detection
The preparation methods of examples 1-3, examples 6-8, examples 11-13 and comparative examples 1-3 are respectively adopted to prepare the koji blocks, the koji blocks are placed at 35 ℃ for fermentation for 72h and then dried for 36h to prepare the fermented feed, the activity of the protease in the fermented feed is determined by adopting the Fulin method, the data is recorded, and the result retains 4 significant digits.
And detection of digestibility
The preparation methods of examples 1-3, examples 6-8, examples 11-13 and comparative examples 1-3 are respectively adopted to prepare the koji blocks, the koji blocks are placed at 35 ℃ for fermentation for 72h and then dried for 36h to prepare the fermented feed, the prepared fermented feed is respectively adopted to feed chicks, the crude protein content in the poultry manure is detected by using the Kjeldahl method 6 days before the feeding of the fermented feed, the crude protein content in the poultry manure is detected by using the Kjeldahl method 12 days after the feeding of the fermented feed, and data are recorded.
TABLE 4 enzyme activity and digestion and absorption rate detection table
Figure BDA0003122781890000121
Figure BDA0003122781890000131
Examples 1-3 in combination with table 4 show that fermented feed prepared from the koji blocks prepared in the present application has higher protease activity and better digestibility.
As can be seen by combining example 1 with examples 6 to 8 and table 4, the fermented feeds prepared in examples 6 to 8 have higher protease activity than example 1 and higher digestibility than example 1; the higher reproduction amount of the clostridium butyricum is proved, and the nutritive value and the digestibility of the fermented feed can be improved.
As can be seen by combining example 6 with examples 11 to 13 and by combining Table 4, the fermented feeds prepared in examples 11 to 13 have higher protease activity than example 6 and higher digestibility than example 6; the modified diatomite and the lactose are matched to increase the number of clostridium butyraldehyde, so that the nutritive value and the digestibility of the fermented feed are improved.
In combination with example 1 and comparative examples 1-3 and table 4, it can be seen that, compared to example 1, the fermented feed prepared in comparative example 1 has a protease activity lower than that of example 1 and a crude protein content in feces higher than that of example 1 after 12 days of feeding, when bacillus subtilis of the same mass is used as a raw material in the comparative example 1 to replace the mold; the mold and the bacillus subtilis are matched, so that amylase, protease, cellulase and other substances can be generated, and the digestibility of the fermented feed is improved.
Comparative example 2 in which yeast of the same mass was substituted for bacillus coagulans in the raw material, compared to example 1, the activity of protease in the fermented feed prepared in comparative example 2 was lower than that in example 1, and the content of crude protein in feces after 12 days of feeding was higher than that in example 1; the yeast and the bacillus coagulans are matched, so that the digestibility and the nutritive value of the fermented feed can be improved.
Comparative example 3 the same mass of bacillus subtilis was used to replace clostridium butyricum in the raw material, compared to example 1, the protease activity of the fermented feed prepared in comparative example 3 is lower than that of example 1, and the content of crude protein in feces after 12 days of feeding is higher than that of example 1; the bacillus subtilis and the clostridium butyricum are matched, so that the digestibility and the nutritive value of the fermented feed can be improved.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.

Claims (7)

1. A koji block for fermented feed, characterized in that the koji block is prepared from the following raw materials in parts by weight: 55-68 parts of yeast block material, 8-16 parts of mixed bacterial powder and 36-54 parts of water; the mixed bacteria powder consists of mould, bacillus subtilis, microzyme, bacillus coagulans and clostridium butyricum with the weight ratio of 0.4-1.6:1:0.4-1.6:0.5-1.5: 0.5-1.5;
the preparation method for the fermented feed koji comprises the following steps:
s1, weighing the clostridium butyricum and the yeast block material with the total weight of 1/8-1/4 and water with the total weight of 1/10-1/6, mixing and stirring to prepare a granular molding material; spraying a film forming solution on the surface of the molding material to prepare a primary mixed material;
s2, weighing mould, bacillus subtilis, saccharomycete and bacillus coagulans, mixing and stirring the mould, the bacillus subtilis and the bacillus coagulans with the rest of the yeast cake materials and the rest of the water, adding the initial mixture prepared in the step S1, continuously stirring, performing compression molding, and fermenting for 10-20 days under the conditions that the temperature is 30-35 ℃ and the humidity is 80-90% to prepare finished yeast cakes;
the film forming solution is prepared by the following method:
weighing 10.8g of wheat gluten protein, 3g of glycerol, 0.5g of Tween 80, 0.5g of guar gum and 102g of ethanol with the mass fraction of 57%, stirring for 5min at the rotating speed of 500r/min, adjusting the pH of the solution to 10 by using sodium hydroxide while stirring, stirring for 30min in a constant-temperature water bath at the temperature of 80 ℃, then carrying out ultrasonic treatment for 15min, and degassing for 10min under the vacuum degree of 0.09MPa to prepare a film forming solution.
2. A koji block for fermented feed according to claim 1, wherein: the mould consists of aspergillus niger and aspergillus oryzae; the yeast comprises Saccharomyces cerevisiae and Candida utilis.
3. A koji block for fermented feed according to claim 1, wherein the koji block material is prepared from raw materials comprising in parts by weight: 15-25 parts of corn flour, 14-24 parts of soybean meal, 46-55 parts of wheat flour, 5-12 parts of molasses and 0.6-1.8 parts of mineral substances.
4. A koji for fermented feed according to claim 3, wherein the mineral substance consists of sodium chloride and dipotassium hydrogen phosphate in a weight ratio of 1: 0.6-1.5.
5. The method of claim 1, wherein the total amount of water in S1 is 1/10-1/6, and the rest of water in S2 is tap water.
6. The method for preparing koji used in fermented feed according to claim 1, wherein modified diatomaceous earth and lactose are added to S1, and the weight ratio of lactose and modified diatomaceous earth to koji material in S1 is 0.02-0.08:0.02-0.15: 1.
7. The method for preparing koji blocks for fermented feed according to claim 6, wherein the modified diatomaceous earth is prepared by the following method:
weighing kieselguhr, placing the kieselguhr into a magnesium sulfate aqueous solution with the mass fraction of 2-8%, wherein the mass ratio of the kieselguhr to the magnesium sulfate aqueous solution is 1:0.8-1.5, stirring, and taking out the kieselguhr adsorbed with magnesium sulfate to prepare the modified kieselguhr.
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