CN113424896B - Fermentation process of bacterial enzyme feed - Google Patents
Fermentation process of bacterial enzyme feed Download PDFInfo
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- CN113424896B CN113424896B CN202110680166.2A CN202110680166A CN113424896B CN 113424896 B CN113424896 B CN 113424896B CN 202110680166 A CN202110680166 A CN 202110680166A CN 113424896 B CN113424896 B CN 113424896B
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
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- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/33—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from molasses
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Abstract
The application relates to the field of microbial fermentation, and particularly discloses a fermentation process of a fungal enzyme feed, which comprises the following steps: s1, preparing yeast blocks, and dissolving the yeast blocks to prepare feed seed liquid; the yeast block is prepared from yeast block materials and mixed bacterial powder; the mixed bacteria powder consists of saccharomycete powder and bacillus subtilis powder; s2, stirring and fermenting the feed seed liquid prepared in the S1 at 28-38 ℃ for 68-78h, and then stirring and drying at 47-58 ℃ for 32-40h to prepare finished bacterial enzyme feed; the bacterial enzyme feed has good storage stability in a higher temperature environment, and even if the animals are not used for eating once and the residual bacterial enzyme feed is exposed to the higher environment temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the animals can still effectively stimulate the feed intake of the animals and improve the digestion and absorption rate after eating the bacterial enzyme feed.
Description
Technical Field
The application relates to the field of microbial fermentation, in particular to a fermentation process of a bacterial enzyme feed.
Background
The microbial enzyme feed is a biological enzyme feed which takes microorganisms and complex enzyme as fermentation agent strains of the biological feed and converts feed raw materials into microbial mycoprotein, bioactive small peptide amino acid, microbial active probiotics and a complex enzyme preparation.
The feed is homogenized and fluffy after being fermented, so that the palatability of the feed is improved, and the feed intake of animals is stimulated; complex macromolecular organic matters such as cellulose, starch, protein and the like in the feed are degraded into micromolecular substances such as monosaccharide, disaccharide, oligosaccharide, amino acid and the like which are easy to digest and absorb by animals to a certain extent, so that the digestibility of the feed is improved; meanwhile, a large amount of microorganism bacterial cells rich in nutrition and useful metabolites are generated and accumulated in the process of fermenting the feed, so that more nutrient substances are provided for animals.
The general fermented feed needs to be stored in a low-temperature dry environment, when the storage temperature is higher than 15 ℃, unstable nutrient substances in the feed begin to lose activity, particularly in a high-temperature environment in summer, the average temperature is higher than 20 ℃, the loss of the nutrient substances in the feed can reach 10% or more, and therefore the feed needs to be placed in the low-temperature dry environment before animals are raised; when animals are raised, a general feeder can adopt a one-time feeding mode according to the daily feed intake of the animals, the feed cannot be used up at one time, and under the condition of summer climate, due to the fact that the environmental temperature is high, nutrient substances in the residual feed easily lose activity under the environment with high temperature, and therefore the feed intake and the digestibility of the animals are easily influenced.
Disclosure of Invention
In order to enable the bacterial enzyme feed to have good storage stability in a higher temperature environment, and enable the nutrient substances in the residual bacterial enzyme feed to still have good biological activity even if the animal is not eaten at one time and the residual bacterial enzyme feed is exposed to the higher environment temperature, the animal can still effectively stimulate the feed intake of the animal and improve the digestibility after eating the feed, the application provides a fermentation process of the bacterial enzyme feed.
The fermentation process of the bacterial enzyme feed adopts the following technical scheme:
a fermentation process of a bacterial enzyme feed comprises the following steps:
s1, preparing yeast blocks, and dissolving the yeast blocks to prepare feed seed liquid; the yeast block is prepared from yeast block materials and mixed bacterial powder; the mixed bacteria powder consists of saccharomycete powder and bacillus subtilis powder;
s2, stirring and fermenting the feed seed liquid prepared in the S1 at 28-38 ℃ for 68-78h, and then stirring and drying at 47-58 ℃ for 32-40h to prepare the finished bacterial enzyme feed.
By adopting the technical scheme, yeast and bacillus subtilis are utilized to prepare the koji blocks, a large amount of thalli are attached to the koji blocks, and the thalli are uniformly dispersed in the feed seed liquid after the koji blocks are dissolved; fermenting the feed seed liquid at 28-38 deg.C for 68-78h, and decomposing the nutrient substances in the yeast block material with yeast, and the metabolite has good resistance.
The bacillus subtilis rapidly grows and breeds in the fermentation process, and then a large amount of spores are generated; drying at 47-58 deg.C for 32-40h, with gradual loss of water, further promoting spore generation in Bacillus subtilis, and drying at this temperature will not inactivate spore.
The resistance of the yeast metabolite is matched with the resistance of bacillus subtilis spores, so that the bacterial enzyme feed has good storage stability, and even if the animal cannot eat the feed at one time and the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of the animal can be effectively stimulated and the digestibility can be improved after the animal eats the feed.
Preferably, the koji block in S1 is prepared by the following method:
preparing mixed bacterium powder, wherein the mixed bacterium powder is prepared by mixing saccharomycete powder, bacillus subtilis powder and clostridium butyricum powder in a weight ratio of 1:0.2-0.7: 0.5-1.5;
II, preparing a yeast block material, mixing the yeast block material, the mixed bacterial powder prepared from the step I and water according to the weight ratio of 6.5:0.5-1.5:3-5, then briquetting and forming, and then fermenting to prepare the yeast block.
By adopting the technical scheme, the yeast, the bacillus subtilis and the clostridium butyricum are matched to prepare mixed bacterial powder, and then the mixed bacterial powder is used for preparing yeast blocks; the surface of the koji block is in direct contact with oxygen, bacillus subtilis grows on the surface of the koji block, oxygen does not exist in the central position of the koji block, clostridium butyricum grows in the central position of the koji block, and facultative anaerobic yeast grows between the surface of the koji block and the central position of the koji block, so that different strains can grow in different positions on the koji block in the same time period, the fermentation time is shortened, and the fermentation cost is saved.
After fermentation of a koji block prepared by matching bacillus subtilis and clostridium butyricum, clostridium butyricum generates a large amount of spores which are endogenous spores and have strong resistance to high temperature and high heat of the external environment; the bacillus subtilis also generates a large amount of spores which have good high-temperature resistance and drying resistance, and the activity of the spores cannot be damaged under the condition of water shortage; the bacillus of clostridium butyricum and bacillus subtilis are matched in a proper proportion, so that the bacterial enzyme feed has better storage stability.
Free oxygen in the intestinal environment can be rapidly consumed after spores of bacillus subtilis enter the intestinal tract, and the intestinal hypoxia environment is formed, so that activity of clostridium butyricum spores is restored, and when the spores of bacillus subtilis and clostridium butyricum are matched, beneficial flora in the intestinal tract is promoted to reproduce, abnormal proliferation of pathogenic bacteria and putrefying bacteria in the intestinal tract can be prevented, the digestion rate of bacteria enzyme feed is improved, and the feed intake of animals is improved.
Preferably, the post-fermentation temperature in the step II is 30-35 ℃, the relative humidity is 80-90%, and the time is 12-18 d.
By adopting the technical scheme, the pressed and formed yeast blocks are fermented for 12-18d under proper conditions,
the bacillus subtilis can grow on the surface of the koji block in a large amount, oxygen in the inner part of the koji block can be absorbed in the growth process of the bacillus subtilis, yeast can breed in a large amount under the aerobic condition in the fermentation process, the yeast can also absorb partial oxygen in the koji block, and the oxygen absorption effect of the layer-by-layer matching of the bacillus subtilis and the yeast ensures that the inner part of the koji block is almost free of oxygen, so that the growth and the breeding of clostridium butyricum are promoted.
Through the fermentation of the formed koji blocks, the koji blocks simultaneously contain a large amount of bacillus subtilis, saccharomycetes and clostridium butyricum at the same time, the spore content is high when the thallus content is high, so that the feed seed liquid can form a large amount of spores in the fermentation process, and the bacterial enzyme feed has good storage stability.
Preferably, the size of the curved block in S1 is 20cm × 15cm × 7 cm.
By adopting the technical scheme, the prepared koji block is a cuboid, the cuboid and the cube have the same volume, the surface area of the cuboid is large, and the koji block is made into the cuboid which can be contacted with oxygen in the external environment in a larger area, so that the growth and the propagation of bacillus subtilis and saccharomycetes on the koji block are promoted.
Through the size of injecing the bent piece, under the prerequisite that bacillus subtilis, the reproduction of growing in a large number of yeasts, oxygen is difficult to permeate inside the bent piece, makes the inside clostridium butyricum of bent piece can grow in a large number and breed under the condition of not contacting with oxygen to make the bent piece that makes when the fermentation, can produce a large amount of bacillus subtilis, yeasts and clostridium butyricum simultaneously at the same time.
Preferably, the koji material in the S1 is prepared from the following raw materials in parts by weight: 15-25 parts of corn flour, 15-25 parts of soybean meal, 45-55 parts of wheat flour, 4-14 parts of molasses and 1-2 parts of mineral substances; the pulverizing fineness of the above materials is 1-3 mm.
By adopting the technical scheme, the corn flour, the soybean meal, the wheat flour, the molasses and the mineral substances are mixed according to a proper proportion, the molasses is used as a bonding material for connecting other materials by utilizing the good bonding effect of the molasses; the appearance of the wheat flour is generally oval, the soybean meal is generally irregular, and the wheat flour is used as a filling material to be filled into the stacking structure of the soybean meal and corn flour particles, so that the internal pores of the koji blocks are reduced, and the internal oxygen of the koji blocks is reduced, thereby promoting the growth and reproduction of clostridium butyricum and yeast.
The corn flour and the soybean meal contain C-source and N-source substances with high content, the corn flour and the soybean meal are used as main materials to provide high-content nutrient substances for the growth and the propagation of the thalli, and the wheat flour contains part of C-source and N-source substances, so that the corn flour, the soybean meal and the wheat flour are matched to further provide high-content nutrient substances for the growth and the propagation of the thalli; the mineral is supplementary material, and the mineral and molasses are matched to provide nutrient substances such as sugar, inorganic salt and the like for the growth and propagation of bacteria; the prepared yeast block material can provide nutrient substances for thalli at different stages of growth and reproduction, so that the growth and reproduction of the thalli are promoted, the thalli generate a large amount of spores, and the bacterial enzyme feed has good storage stability.
Preferably, the stirring speed in the S2 fermentation process is 0-8r/min, and the stirring speed in the drying process is 8-15 r/min.
By adopting the technical scheme, the stirring speed of the feed seed liquid in the fermentation process is limited, so that the bacillus subtilis and the saccharomycetes in the feed seed liquid can be better contacted with oxygen and nutrient substances in the yeast block materials in the growth and reproduction processes to promote the growth and reproduction of the bacillus subtilis and the saccharomycetes, and the growth and reproduction of the thalli are not easily inhibited at a lower stirring speed, so that the thalli in the feed seed liquid can generate spores to the maximum extent in the fermentation process to improve the storage stability of the microbial enzyme feed.
Through the stirring speed of injecing among the drying process for moisture rapid draing, the rapid loss of moisture makes the spore generate fast, thereby further makes the thallus produce the spore, with the storage stability that improves fungus enzyme feed.
Preferably, the feed seed solution in S2 is fermented for 72h at the temperature of 35 ℃ and the rotation speed of 4r/min, and then dried for 36h at the temperature of 55 ℃ and the rotation speed of 12 r/min.
By adopting the technical scheme, the fermentation time is 72 hours and the drying time is 36 hours, so that a large amount of spores are formed, the content of thallus metabolites is high, the fungus enzyme feed has good storage stability, and has good biological activity even after being stored for a long time, and the feed intake of animals can be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the fungus enzyme feed.
Preferably, the temperature of the hot air is increased at a temperature increasing rate of 1-3 ℃/2min in the S2.
By adopting the technical scheme, the temperature rising speed is controlled and the temperature rising is matched with hot air, so that the moisture drying is accelerated, and the further spore generation of thalli is promoted, thereby improving the storage stability of the bacterial enzyme feed.
Preferably, the S1 is used for preparing the yeast blocks, the yeast blocks are dissolved by water and then mixed with the bamboo powder, and the mass ratio of the yeast blocks, the water and the bamboo powder is 1:7-10:0.05-0.2, so that the feed seed liquid is prepared.
By adopting the technical scheme, the bacillus subtilis absorbs nutrient substances in the koji block in the process of initial growth and propagation and supplies the initial thalli with the nutrient substances absorbed, new thalli are continuously generated along with the continuous increase of the growth and propagation amount, part of newly generated bacillus subtilis is gradually attached to the surface of rough bamboo powder, the bamboo powder is used as a supporting matrix, and the nutrient substances in the koji block are captured by virtue of the porous rough structure of the bamboo powder, so that the newly grown thalli can be promoted to better absorb the nutrient substances, and the new thalli are continuously propagated and generated; the bamboo powder and the yeast blocks are matched, so that the thalli in different growth stages can be promoted to uniformly absorb nutrient substances, the thalli can continuously grow and reproduce, the number of the thalli is increased, the spore content is further increased, and the storage stability of the bacterial enzyme feed is improved.
Preferably, the bamboo powder is modified bamboo powder, and the modified bamboo powder is prepared by the following method:
and spraying the gelatin solution on the surface of the bamboo powder, wherein the weight ratio of the gelatin solution to the bamboo powder is 1:3-5.5, and drying to obtain the modified bamboo powder.
By adopting the technical scheme, the gelatin is attached to the surface of the bamboo powder, the gelatin solution is dried to form the gelatin film, and after the bamboo powder and the koji blocks are prepared into the feed seed liquid, the gelatin film can prevent the bamboo powder from absorbing the moisture in the seed liquid so as to ensure that the thalli absorb the moisture and water-soluble nutrient substances and ensure the growth and reproduction of the thalli.
The attachment of the gelatin film increases the outer surface area of the bamboo powder, so that new thalli can be attached to the surface of the modified bamboo powder conveniently, metabolites of acid substances can be generated along with the growth and the propagation of the thalli, the gelatin film is easy to decompose the acid substances, and small molecular substances such as amino acid and the like can be generated after the decomposition of protein in the gelatin film, so that nutrient substances are provided for the new thalli attached to the surface of the modified bamboo powder, the growth and the propagation of the new thalli are promoted, the thalli in different growth stages can grow better, the spore content is further increased by increasing the number of the thalli, and the storage stability of the enzyme feed is improved.
In summary, the present application has the following beneficial effects:
1. the resistance of the yeast metabolite is matched with the resistance of bacillus subtilis spores, so that the bacterial enzyme feed has good storage stability, and even if the animal cannot eat the feed at one time and the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of the animal can be effectively stimulated and the digestibility can be improved after the animal eats the feed.
2. Preparing mixed bacterial powder by matching saccharomycetes, bacillus subtilis and clostridium butyricum, and preparing a koji block by using the mixed bacterial powder; different strains can grow at different positions on the koji block in the same time period, so that the fermentation time is shortened, and the fermentation cost is saved.
3. Fermenting for 108h to ensure that the yeast is completely fermented to completely generate metabolites, wherein the completely fermented yeast does not try to obtain nutrients in animal intestines after entering the animal intestines, and more metabolites of natural nutrients, such as UGF, enzymes and the like, are produced after complete metabolism, so that more nutrients can be provided for the animals.
4. The bacillus subtilis and the clostridium butyricum are matched, so that the spore content in the bacterial enzyme feed can be further improved, and the resistance of the bacterial enzyme feed can be further improved.
5. The bamboo powder, the yeast blocks and the water are matched, so that the growth and the propagation of the bacillus subtilis are further promoted, the quantity of the bacteria is increased, the spore content is further increased, and the storage stability of the bacterial enzyme feed is improved.
Detailed Description
The present application will be described in further detail with reference to examples.
Preparation example of modified bamboo powder
The bamboo powder in the following raw materials is purchased from silicone mining company Limited in Gao county, and the specification is 80 meshes; other raw materials and equipment are all sold in the market.
Preparation example 1: the modified bamboo powder is prepared by the following method:
preparing a gelatin aqueous solution with the mass fraction of 2%, spraying the gelatin aqueous solution on the surface of the bamboo powder by using a film coating machine, wherein the weight ratio of the gelatin aqueous solution to the bamboo powder is 1:4.2, and drying to obtain modified bamboo powder; sieving bamboo powder with 80 mesh sieve.
Preparation example 2: the difference between the preparation example and the preparation example 1 is that:
the weight ratio of the gelatin water solution to the bamboo powder is 1: 3.
Preparation example 3: the difference between the preparation example and the preparation example 1 is that:
the weight ratio of the gelatin water solution to the bamboo powder is 1: 5.5.
Preparation example of koji
The yeast in the following raw materials is purchased from saccharomyces cerevisiae produced by Qingdao root biotechnology group limited; the bacillus subtilis is purchased from bacillus subtilis powder produced by Qingdao root biotechnology group Limited, and the model is CT-10; the clostridium butyricum is purchased from clostridium butyricum powder produced by Qingdao root biotechnology group limited; other raw materials and equipment are all sold in the market.
Preparation example 4: the yeast block is prepared by the following method:
weighing 1kg of saccharomycete powder and 0.5kg of bacillus subtilis powder, and mixing and stirring to prepare mixed bacteria powder;
II, crushing corn flour, soybean meal and wheat flour to the fineness of 2mm, weighing 20kg of corn flour, 20kg of soybean meal, 50kg of wheat flour, 8.5kg of molasses and 1.5kg of mineral substances, and mixing to obtain a yeast cake material; the mineral substance comprises sodium chloride and sodium dihydrogen phosphate at a weight ratio of 1: 1; weighing 65kg of yeast block materials, placing the materials in a mixer, stirring for 2min at the rotating speed of 500r/min, adding 10kg of mixed bacterial powder and 44kg of water, continuing stirring for 15min, then briquetting and forming by using a forming machine to obtain yeast blanks, and carrying out after-fermentation on the yeast blanks for 15d under the conditions of 32 ℃ and 85% relative humidity to obtain yeast blocks; the block size is 20cm × 15cm × 7 cm.
Preparation example 5: the difference between the preparation example and the preparation example 4 is that:
weighing 1kg of saccharomycete powder, 0.5kg of bacillus subtilis powder and 1kg of clostridium butyricum powder, mixing and stirring to obtain mixed bacteria powder.
Preparation example 6: the difference between the preparation example and the preparation example 4 is that:
weighing 1kg of saccharomycete powder, 0.2kg of bacillus subtilis powder and 0.5kg of clostridium butyricum powder, mixing and stirring to obtain mixed powder.
Preparation example 7: the difference between the preparation example and the preparation example 4 is that:
weighing 1kg of saccharomycete powder, 0.7kg of bacillus subtilis powder and 1.5kg of clostridium butyricum powder, mixing and stirring to obtain mixed powder.
Preparation example 8: the difference between the preparation example and the preparation example 4 is that:
II, crushing corn flour, soybean meal and wheat flour to the fineness of 1mm, and weighing 15kg of corn flour, 15kg of soybean meal, 55kg of wheat flour, 4kg of molasses and 1kg of mineral matter to prepare a yeast cake material; the mineral substance comprises sodium chloride and sodium dihydrogen phosphate at a weight ratio of 1: 1; weighing 65kg of yeast block materials, placing the materials in a mixer, stirring for 2min at the rotating speed of 500r/min, adding 5kg of mixed bacterial powder and 30kg of water, continuing stirring for 15min, then briquetting and forming by using a forming machine to obtain yeast blanks, and carrying out after-fermentation on the yeast blanks for 12d under the conditions of 30 ℃ and 80% of relative humidity to obtain yeast blocks; the block size is 20cm × 15cm × 7 cm.
Preparation example 9: the difference between the preparation example and the preparation example 4 is that:
II, crushing corn flour, soybean meal and wheat flour to the fineness of 3mm, and weighing 25kg of corn flour, 25kg of soybean meal, 45kg of wheat flour, 14kg of molasses and 2kg of mineral substances to prepare a yeast cake material; the mineral substance comprises sodium chloride and sodium dihydrogen phosphate at a weight ratio of 1: 1; weighing 65kg of yeast block materials, placing the materials in a mixer, stirring for 2min at the rotating speed of 500r/min, adding 15kg of mixed bacterial powder and 50kg of water, continuing stirring for 15min, then briquetting and forming by using a forming machine to obtain yeast blanks, and carrying out after-fermentation on the yeast blanks for 18d under the conditions of 35 ℃ and relative humidity of 90% to obtain yeast blocks; the block size is 20cm × 15cm × 7 cm.
Examples
The following raw materials and equipment are all commercially available.
Example 1: a fermentation process of a bacterial enzyme feed comprises the following steps:
s1, weighing 140kg of the koji prepared in preparation example 4, 1260kg of tap water, and placing in a container with a volume of 2m3Stirring in a stirring tank until the koji is completely dissolved to obtain feed seed liquid; the temperature of tap water is 30-35 ℃;
s2, fermenting the feed seed liquid prepared in the S1 for 72 hours at the temperature of 35 ℃ and the rotating speed of 4r/min, and then drying for 36 hours at the temperature of 55 ℃ and the rotating speed of 12r/min, wherein the heating rate is 2 ℃/2min, and the heating mode is hot air heating, so that the finished product of the bacterial enzyme feed is prepared.
Example 2: the present embodiment is different from embodiment 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 78 hours at the temperature of 28 ℃ and the rotating speed of 0r/min, and then drying the feed seed liquid for 32 hours at the temperature of 47 ℃ and the rotating speed of 8r/min, wherein the heating rate is 1 ℃/2min, and the heating mode is hot air heating, so that the finished product of the bacterial enzyme feed is prepared.
Example 3: the present embodiment is different from embodiment 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 68 hours at the temperature of 38 ℃ and the rotating speed of 8r/min, and then drying the feed seed liquid for 40 hours at the temperature of 58 ℃ and the rotating speed of 15r/min, wherein the heating rate is 2 ℃/2min, and the heating mode is hot air heating, so that the finished product of the bacterial enzyme feed is prepared.
Example 4: the present embodiment is different from embodiment 1 in that:
the koji blocks prepared in preparation example 5 were selected.
Example 5: the present embodiment is different from embodiment 1 in that:
the koji blocks prepared in preparation example 6 were selected.
Example 6: the present embodiment is different from embodiment 1 in that:
the koji blocks prepared in preparation example 7 were selected.
Example 7: the present embodiment is different from embodiment 1 in that:
the koji blocks prepared in preparation example 8 were selected.
Example 8: the present embodiment is different from embodiment 1 in that:
the koji blocks prepared in preparation example 9 were selected.
Example 9: this embodiment is different from embodiment 4 in that:
s1, weighing 140kg of the koji prepared in preparation example 5, 1260kg of tap water and 14kg of bamboo powder, and placing the materials in a container with a volume of 2m3Stirring in a stirring tank until the koji is completely dissolved to obtain the feed seed liquid.
Example 10: this embodiment is different from embodiment 4 in that:
s1, weighing 140kg of the koji prepared in preparation example 5, 980kg of tap water and 7kg of bamboo powder, and placing the materials in a space of 2m3Stirring in a stirring tank until the koji is completely dissolved to obtain the feed seed liquid.
Example 11: this embodiment is different from embodiment 4 in that:
s1, weighing140kg of the koji prepared in preparation example 5, 1400kg of tap water, 28kg of bamboo powder were placed in a space of 2m3Stirring in a stirring tank until the koji is completely dissolved to obtain the feed seed liquid.
Example 12: the present embodiment is different from embodiment 9 in that:
the bamboo powder prepared in preparation example 1 is selected as the bamboo powder.
Example 13: the present embodiment is different from embodiment 9 in that:
the bamboo powder prepared in preparation example 2 is selected as the bamboo powder.
Example 14: the present embodiment is different from embodiment 9 in that:
the bamboo powder prepared in preparation example 3 is selected as the bamboo powder.
Example 15: the present embodiment is different from embodiment 1 in that:
the yeast blocks are not post-fermented in the preparation process.
Example 16: the present embodiment is different from embodiment 1 in that:
the raw materials of the yeast block material are corn flour with the same mass instead of wheat flour.
Example 17: the present embodiment is different from embodiment 1 in that:
the block size is 15cm × 15cm × 15 cm.
Comparative example
Comparative example 1: this comparative example differs from example 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 72 hours at the temperature of 35 ℃ and the rotating speed of 4r/min to prepare the finished product of the bacterial enzyme feed.
Comparative example 2: this comparative example differs from example 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 108 hours at the temperature of 35 ℃ and the rotating speed of 4r/min to prepare the finished product of the bacterial enzyme feed.
Comparative example 3: this comparative example differs from example 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 36 hours at the temperature of 35 ℃ and the rotating speed of 4r/min, and then drying for 72 hours at the temperature of 55 ℃ and the rotating speed of 12r/min, wherein the heating rate is 2 ℃/2min, and the heating mode is hot air heating, so that the finished product of the bacterial enzyme feed is prepared.
Comparative example 4: this comparative example differs from example 1 in that:
s2, fermenting the feed seed liquid prepared in the S1 for 60 hours at the temperature of 35 ℃ and the rotating speed of 4r/min, and then drying for 48 hours at the temperature of 55 ℃ and the rotating speed of 12r/min at the heating rate of 2 ℃/2min in a hot air heating mode to prepare the finished product of the bacterial enzyme feed.
Performance test
1. Detection of microbial and spore numbers
The preparation methods of examples 1-17 and comparative examples 1-4 are respectively adopted to prepare the fungal enzyme feed, the method of GB478915-2010 food microbiology inspection of mould and yeast counting is adopted, the number of the yeast in the koji is detected after the preparation of the koji, and the number of the yeast is respectively detected after the fungal enzyme feed is fermented and dried (the finished product is obtained after the yeast is dried).
TABLE 1 Yeast number test Table
Preparing enzyme feeds by adopting the preparation methods of examples 1-17 and comparative examples 1-4 respectively, detecting the number of bacillus subtilis in the koji blocks after the preparation of the koji blocks by adopting the detection of bacillus subtilis in a microbial preparation for feeding GB/T26428-2010, and detecting the number of bacillus subtilis in the koji blocks after the enzyme feeds are fermented and dried respectively; the number of the bacillus subtilis in the finished bacterial enzyme feed is multiplied by 93 percent, namely the number of the spores of the bacillus subtilis in the finished bacterial enzyme feed (the number of the spores is reserved by 3 as an effective number); comparative examples 1-2 without drying procedure, the number of Bacillus subtilis after fermentation multiplied by 93% was the number of spores of Bacillus subtilis in the finished bacterial enzyme feed.
TABLE 2 Bacillus subtilis quantity test table
Respectively adopting the preparation methods of examples 4-17 and comparative examples 1-4 to prepare the bacterial enzyme feed, adopting a DB41/T1728-2018 feed additive clostridium butyricum determination microbiological method, detecting the number of clostridium butyricum in a koji block after the preparation of the koji block is finished, and respectively detecting the number of clostridium butyricum after the bacterial enzyme feed is fermented and dried; the number of clostridium butyricum in the finished bacterial enzyme feed is multiplied by 91 percent, namely the number of spores of clostridium butyricum in the finished bacterial enzyme feed (the number of spores is reserved 3 as an effective number); comparative examples 1-2 without drying step, the number of clostridium butyricum after fermentation multiplied by 91% was the number of spores of clostridium butyricum in the finished bacterial enzyme feed.
TABLE 3 Clostridium butyricum quantity test Table
2. Enzyme activity detection
The preparation methods of examples 1-17 and comparative examples 1-4 are respectively adopted to prepare the bacterial enzyme feed, the activity of the protease in the bacterial enzyme feed is determined by adopting the Fulin method, the data is recorded, and the data retains 4 significant figures.
3. Feed intake detection
Respectively adopting the preparation methods of examples 1-17 and comparative examples 1-4 to prepare the bacterial enzyme feed, respectively adopting the prepared bacterial enzyme feed to feed chicks, respectively adopting the environment temperature of the bacterial enzyme feed after opening the bags to be 38 ℃ and the relative humidity to be 55%, respectively feeding the chicks after storing for 6h and 12h, wherein the weight of the chicks is 600g, and recording the daily feed intake.
4. Digestibility detection
Respectively adopting the preparation methods of examples 1-17 and comparative examples 1-4 to prepare the bacterial enzyme feed, respectively adopting the prepared bacterial enzyme feed to feed chicks, enabling the environment temperature of the bacterial enzyme feed after opening the bags to be 38 ℃ and the relative humidity to be 55%, feeding the chicks after storing for 6h, continuously feeding the chicks for 12 days, detecting the crude protein content in the poultry manure by adopting a Kjeldahl method on the 12 th day, and recording data.
TABLE 4 enzyme activity, feed intake and digestion and absorption rate test table
Combining with the examples 1-3 and tables 1, 2, 3 and 4, it can be seen that the yeast and bacillus subtilis in the koji block are higher in number, the dried thallus is still higher in number, the transformed spore content is also higher, meanwhile, the protease activity is higher, the animal feed intake is higher, and the digestion and absorption rate of the fungal enzyme feed is better; the yeast and the bacillus subtilis are fermented and dried to prepare the yeast block, the protein in the raw material is decomposed into the micromolecular nutrient substance by the yeast, and the bacillus subtilis generates a large amount of spores, so that the prepared finished bacterial enzyme feed utilizes the high resistance of the spores to be matched with the good resistance of the metabolite of the yeast, the bacterial enzyme feed has good storage stability, and has good biological activity even after being stored at high temperature for a long time, and the feed intake of animals can be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the feed.
Combining example 1 and examples 4-6 with tables 1, 2, 3 and 4, it can be seen that the mixed powder of examples 4-6 is prepared by mixing saccharomycete powder, bacillus subtilis powder and clostridium butyricum powder, compared with example 1, the yeast and bacillus subtilis quantities in the koji blocks of examples 4-6 are smaller than that in example 1, and the yeast and bacillus subtilis quantities in the fermentation and drying of examples 4-6 are smaller than that in example 1, and the bacillus subtilis quantities in the fermentation and drying of examples 4-6 are smaller than that in example 1, so that the clostridium butyricum content is higher; the addition of the clostridium butyricum reduces the proportion of the yeast and the bacillus subtilis to a certain extent, so that the quantity of the yeast and the bacillus subtilis is influenced, but spores in the bacillus subtilis are matched with spores in the clostridium butyricum, the content of the spores in the bacterial enzyme feed is improved, and the storage stability of the bacterial enzyme feed is improved; even if the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can still be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the feed.
The protease activity of the examples 4-6 is less than that of the example 1, but the difference between the 1d and 3d feed intake of the fungal enzyme feed prepared in the examples 4-6 is less than that of the example 1, and the crude protein content in the animal manure of the examples 4-6 is greater than that of the example 1; the spores of the clostridium butyricum and the spores of the bacillus subtilis are matched, so that the bacterial enzyme feed has good storage stability, and even if the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can be effectively stimulated and the digestibility can be improved after the animals eat the feed.
Example 1 in combination with examples 7-8 and tables 1, 2, 3, and 4, it can be seen that the yeast and bacillus subtilis numbers in the koji of example 7 are less than those in example 1, and the yeast and bacillus subtilis numbers after fermentation and drying of example 7 are less than those in example 1, and the bacillus subtilis numbers in example 7 are less than those in example 1; the fermentation cake has the advantages that the number of clostridium butyricum is high, the number of clostridium butyricum is high after fermentation and drying, and the number of spores is also high, so that the contents of corn flour, soybean meal and wheat flour are low, the growth of each thallus is easily influenced, and the number of the thallus and the yield of the spores are easily influenced.
Example 7 protease activity was lower than example 1, and the difference between the feed intake of 1d and the feed intake of 3d in the fungal enzyme feed prepared in example 7 was less than that in example 1; indicating that the number of thalli is reduced, the activity of protease is reduced, and the feed intake and the digestion and absorption rate of animals are influenced.
The number of yeasts and bacillus subtilis in the koji block of the example 8 is larger than that in the koji block of the example 1, and the number of yeasts and bacillus subtilis in the koji block of the example 8 is larger than that in the koji block of the example 1 after fermentation and drying, and the number of spores of the bacillus subtilis of the example 8 is larger than that in the koji block of the example 1; the fermentation cake has the advantages that the number of clostridium butyricum is high, the number of clostridium butyricum is high after fermentation and drying, and the number of spores is also high, so that the contents of corn flour, soybean meal and wheat flour are high, the growth of bacteria can be promoted to a certain extent due to the high content of nutrient substances, and the number of bacteria and the yield of spores are easily influenced.
Example 8 protease activity was higher than example 1, and the difference between the feed intake of 1d and the feed intake of 3d in the fungal enzyme feed prepared in example 8 was less than that in example 1; the increase of the number of the thalli, the enhancement of the activity of the protease and the improvement of the feed intake of the animals are proved.
By combining the example 4 and the examples 9 to 11 and combining the tables 1, 2, 3 and 4, it can be seen that the amount of yeasts, bacillus subtilis and clostridium butyricum in the koji blocks of the examples 9 to 11 is larger than that of the example 4, the amount of spores of the bacillus subtilis and clostridium butyricum after fermentation and drying is larger than that of the example 4, the amount of spores of the bacillus subtilis and clostridium butyricum is also larger than that of the example 4, the protease activity is larger than that of the example 4, the difference between the 1d feed intake and the 3d feed intake of the fungal enzyme feed is smaller than that of the example 4, and the content of crude protein in excrement is lower than that of the example 4; the bacillus subtilis, the clostridium butyricum, the saccharomycetes and the bamboo powder are matched, so that the thalli in different growth stages can be promoted to uniformly absorb nutrient substances, the thalli can continuously grow and propagate, the number of the thalli is increased, the spore content is further increased, the storage stability of the bacterial enzyme feed is improved, even if the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can be effectively stimulated and the digestibility is improved after the animals eat the bacterial enzyme feed.
By combining example 9 and examples 12-14 with tables 1, 2, 3 and 4, it can be seen that the amount of yeasts, bacillus subtilis and clostridium butyricum in the koji blocks is greater than that in example 9, the amount of yeasts, bacillus subtilis and clostridium butyricum after fermentation and drying is greater than that in example 9, the number of spores of bacillus subtilis and clostridium butyricum is greater than that in example 9, the protease activity is greater than that in example 9, the difference between the 1d feed intake and the 3d feed intake of the fungal enzyme feed is smaller than that in example 9, and the content of crude protein in feces is lower than that in example 9 when the bamboo powder is added into the raw materials in examples 12-14; the bacillus subtilis, the clostridium butyricum, the saccharomycetes and the modified bamboo powder are matched, the barrier property of the gelatin film is utilized to be matched with the nutritional effect of protein in gelatin, the uniform absorption of nutrients by bacteria can be further promoted, the quantity of the bacteria is increased, the spore content is further increased, the storage stability of the bacterial enzyme feed is further improved, even if the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrients in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can still be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the bacterial enzyme feed.
Example 1 in combination with examples 15-17 and tables 1, 2, 3 and 4, it can be seen that in example 15, compared to example 1, in the process of preparing koji blocks without after-fermentation, the fungal enzyme feed prepared in example 15 has a lower amount of each bacterial cell than in example 1 after fermentation and drying, and the amount of each bacterial cell in koji blocks is also lower than in example 1, the protease activity is lower than in example 1, the feed intake is lower than in example 1, and the content of crude protein in feces is higher than in example 1; the post-fermentation of the yeast blocks can improve the quantity of each thallus, thereby improving the spore content, and improving the digestibility, the nutrient content and the feed intake of the fungal enzyme feed.
Example 16 in the koji material raw material, the same mass of corn flour is used to replace the wheat flour, compared with example 1, the bacterial enzyme feed prepared in example 16 has the bacterial cell number lower than that of example 1 after fermentation or drying, the bacterial cell number in the koji is lower than that of example 1, the protease activity is lower than that of example 1, the feed intake is lower than that of example 1, and the crude protein content in the excrement is higher than that of example 1; the wheat flour, the corn flour and the soybean meal are matched, the internal pores of the koji blocks can be reduced through the stacking structure, the growth and the propagation of clostridium butyricum and saccharomycetes are promoted, the quantity of bacteria is increased, the spore content is increased, even if the residual bacterial enzyme feed is exposed to higher environmental temperature, the nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the feed.
Example 17 the size of the koji block is 15cm × 15cm × 15cm, compared with example 1, the bacterial enzyme feed prepared in example 17 has the bacterial cell number lower than that of example 1 after fermentation and drying, the bacterial cell number in the koji block is lower than that of example 1, the protease activity is lower than that of example 1, the feed intake is lower than that of example 1, and the crude protein content in excrement is higher than that of example 1; the yeast blocks prepared by 20cm multiplied by 15cm multiplied by 7cm can better enable yeast, bacillus subtilis and clostridium butyricum to propagate higher amount of thalli at the same time, so that the spore content of the bacterial enzyme feed is improved, even if the residual bacterial enzyme feed is exposed to higher environmental temperature, nutrient substances in the residual bacterial enzyme feed still have good biological activity, and the feed intake of animals can still be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the feed.
It can be seen from the combination of example 1 and comparative examples 1 to 4 and the combination of tables 1, 2, 3 and 4 that the fermentation time of 72h is matched with the drying time of 36h, so that the yeast and bacillus subtilis have higher thallus content and the bacillus subtilis has higher bacillus subtilis spore content, the fungal enzyme feed has good storage stability in a higher temperature environment, and even if the residual fungal enzyme feed is exposed to a higher environmental temperature, the nutrient substances in the residual fungal enzyme feed still have good biological activity, and the feed intake of animals can be effectively stimulated and the digestion and absorption rate can be improved after the animals eat the feed.
5. Detection of storage time and deterioration
The preparation methods of the examples 1-3 and the comparative example 2 are respectively adopted to prepare the bacterial enzyme feed, the storage time is more than or equal to 8 months, and the bacterial enzyme feed can not be rotten and deteriorated after being placed for 5 days at the temperature of 38-40 ℃ and the relative humidity of 55%; the mixed fermented feed produced by comparative example 2 and commercial Jinan Xin Ying chemical Co Ltd is deteriorated when left for 5 days at 38-40 ℃ and 55% relative humidity.
The bacterial enzyme feed prepared by the method has longer shelf life, does not decay and deteriorate even if placed in an open place, and still can keep the taste and nutrient substances of the bacterial enzyme feed.
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 (6)
1. A fermentation process of a bacterial enzyme feed is characterized by comprising the following steps:
s1, preparing yeast blocks, and dissolving the yeast blocks to prepare feed seed liquid; the yeast block is prepared from yeast block materials and mixed bacterial powder; the mixed bacteria powder consists of saccharomycete powder and bacillus subtilis powder; the size of the bent block is 20cm multiplied by 15cm multiplied by 7 cm;
s2, stirring and fermenting the feed seed liquid prepared in the S1 at 28-38 ℃ for 68-78h, and then stirring and drying at 47-58 ℃ for 32-40h to prepare finished bacterial enzyme feed;
the yeast block in S1 is prepared by the following method:
preparing mixed bacterium powder, wherein the mixed bacterium powder is prepared by mixing saccharomycete powder, bacillus subtilis powder and clostridium butyricum powder in a weight ratio of 1:0.2-0.7: 0.5-1.5;
II, preparing a yeast block material, mixing the yeast block material, the mixed bacterial powder prepared from the step I and water according to the weight ratio of 6.5:0.5-1.5:3-5, then briquetting and forming, and then fermenting to prepare the yeast block; the post-fermentation temperature is 30-35 deg.C, relative humidity is 80-90%, and the time is 12-18 d;
the yeast block material is prepared from the following raw materials in parts by weight: 15-25 parts of corn flour, 15-25 parts of soybean meal, 45-55 parts of wheat flour, 4-14 parts of molasses and 1-2 parts of mineral substances; the pulverizing fineness of the above materials is 1-3 mm.
2. The fermentation process of a fungal enzyme feed as claimed in claim 1, wherein the stirring speed in the S2 fermentation process is 0-8r/min, and the stirring speed in the drying process is 8-15 r/min.
3. The fermentation process of a fungal enzyme feed as claimed in claim 2, wherein the feed seed liquid in S2 is fermented at 35 ℃ and 4r/min for 72h, and then dried at 55 ℃ and 12r/min for 36 h.
4. The fermentation process of a fungal enzyme feed as claimed in claim 1, wherein the temperature of the hot air is increased at a rate of 1-3 ℃/2min in the step of S2.
5. The fermentation process of a fungal enzyme feed as claimed in claim 1, wherein S1 is used for preparing a koji block, the koji block is dissolved in water and then mixed with bamboo powder, and the feed seed liquid is prepared by the mass ratio of the koji block, the water and the bamboo powder being 1:7-10: 0.05-0.2.
6. The fermentation process of the bacterial enzyme feed as claimed in claim 5, wherein the bamboo powder is modified bamboo powder, and the modified bamboo powder is prepared by adopting the following method:
and spraying the gelatin solution on the surface of the bamboo powder, wherein the weight ratio of the gelatin solution to the bamboo powder is 1:3-5.5, and drying to obtain the modified bamboo powder.
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| CN106578458A (en) * | 2016-12-05 | 2017-04-26 | 刁海波 | Growth promoting feed for sheep in weaning period |
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| CN102220261A (en) * | 2011-04-29 | 2011-10-19 | 济南海华生物科技有限公司 | Preparation and use of bacillus subtilis and clostridium butyricum composite bacterial preparation |
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