CN103262949A - Biological enzyme protein feed additive and preparation method thereof - Google Patents
Biological enzyme protein feed additive and preparation method thereof Download PDFInfo
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Abstract
The invention provides a biological enzyme protein feed additive and a preparation method thereof, belonging to the field of feeds. The preparation method comprises the following steps of: respectively culturing under different conditions by utilizing the different characteristics of beer yeast, pediococcus acidilactici and bacillus licheniformis to prepare different strains; then, uniformly connecting 2% of beer yeast strains, 2% of pediococcus acidilactici strains and 1% of bacillus licheniformis strains to a blended aniseed culture medium to mix and carry out submerged fermentation; and drying, crushing, checking and packaging to obtain the product provided by the invention. By using the biological enzyme protein feed additive, the digestion and utilization ratios of a feed can be increased, the anti-nutritional factors in the feed can be degraded, the hyposecretion of the endogenous enzyme of a young animal can be solved, the growth and development of an animal can be promoted, the immunity of the animal can be enhanced, the consumption of antibiotic drugs in feeding animals can be effectively reduced, the faecal phosphorus pollution to the environment can be reduced, the environment pollution can be reduced, and the feed cost can be reduced, so that the biological enzyme protein feed additive has wide market prospects.
Description
Technical field:
The invention belongs to field of fodder, relate in particular to additive of protein feed.
Background technology:
Protein is one of essential three major nutrient of human life activity, and the main source of protein is meat, egg, milk and bean food, and generally speaking, the protein that comes from animal has higher quality, contains sufficient essential amino acid.Obtain and can substantially all rely on for the animal that people eat to culture.Therefore livestock and poultry cultivation and people's health is more and more closely bound up, in order to prevent and treat Animal diseases, the normal antibiotic product that uses is guaranteed animal health in the modern livestock and poultry cultivation process, but excessive antibiotic use not only causes the quality of animal flesh goods and accessory substance thereof to descend and the enhancing of germ patience, and the transmission by food chain and accumulate and cause human health to be subjected to the serious threat of antibiotic problem.The raw material rise causes feeding cost to increase day by day in addition, develop a kind of efficiency of feed utilization that can improve the fowl poultry in a short time, reduce feeding cost, improve feedstuff-meat ratio and breeding performonce fo animals, reduce antibiotic dosage, reduce excrement phosphorus to the pollution of environment, produce the feed addictive of healthy and safe meat products and become the extremely urgent demand of industry of raising thereby strengthen animal physique.Therefore, biological enzyme technology more and more is subjected to people's attention in the application of feedstuff industry.
Summary of the invention:
The invention provides a kind of biology enzyme additive of protein feed and preparation method thereof, described biology enzyme protein feed is according to brewer's yeast, the lactic acid sheet bacterium, the bacillus licheniformis different qualities, adopt different condition to cultivate respectively and be prepared into different strain separately, mix the fermentation of dark pond then, be specially, in proportion with the saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification evenly insert in the modulated aniseed culture medium that makes, the dress pond, behind the scraper constant temperature culture certain hour, drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package; Described three kinds of bacterial classifications add toatl proportion in the aniseed culture medium be 5%, and wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.
Described three kinds of bacterial classifications add toatl proportion in the aniseed culture medium be 5%, and wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.
The saccharomyces cerevisiae strain preparation: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, and switching goes into to be added with in the fermentation tank of fermentation medium, and the control temperature is 28～30 ℃, ventilates early stage to cultivating 16 hours, and throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification.Fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
Lactic acid sheet bacterial preparation: the inclined-plane is cultivated lactic acid sheet bacterium seed liquor and is transferred in the fermentation tank, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa incubation time 36-48 hour, is prepared into lactic acid sheet bacterial.Fermentation tank culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
Bacillus licheniformis strain preparation, inclined-plane are cultivated the bacillus licheniformis seed liquor and are transferred in the fermentation tank, and 28～32 ℃ of 150r/m constant temperature constant speed were cultivated 36-48 hour, were prepared into the bacillus licheniformis bacterial classification.The fermentation tank prescription is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
Mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 48～72 hours, drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
Beneficial effect:
The present invention adopts bacillus licheniformis, lactic acid sheet bacterium, three kinds of bacterial classifications of saccharomyces cerevisiae to ferment, can improve the digestive utilization ratio of feed, ANFs in the degraded feed replenishes young animal endogenous enzymes hyposecretion, promotes animal growth, strengthen the immunity of animal, effectively reduce the use amount of antibiotics in letting animals feed, reduce excrement phosphorus to the pollution of environment, reduce environmental pollution, reduce feed cost, thereby have vast market prospect.
The specific embodiment:
Use this bacterial strain of saccharomyces cerevisiae (Saccharomyces cerevisiae) to buy in Chinese industrial microorganism fungus kind preservation administrative center address: No. 32, Xiaoyun Road, Chaoyang District, Beijing City among the present invention; Postcode: 100027; Deposit number is CICC31362.This bacterial strain of bacillus licheniformis (Bacillus licheniformis) is bought in Chinese industrial microorganism fungus kind preservation administrative center, address: No. 32, Xiaoyun Road, Chaoyang District, Beijing City; Postcode: 100027; Deposit number is CICC10037.
The saccharomyces cerevisiae strain preparation:
The saccharomyces cerevisiae strain preparation: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, and switching goes into to be added with in the fermentation tank of fermentation medium, and the control temperature is 28～30 ℃, ventilates early stage to cultivating 16 hours, and throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification.Fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
Lactic acid sheet bacterium (Lactobacillus) strain preparation:
1. first order seed is cultivated: lactic acid sheet bacterium slant strains is inserted in 500 ml shake flasks 100 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
2. secondary seed is cultivated: first order seed is inserted 1000 milliliters of secondary seeds according to 10% inoculum concentration shake in the bottle, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle 1000 milliliters of culture medium loading amounts, 100 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa incubation time 36-48 hour, cultivates and finishes bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
The bacillus licheniformis strain preparation:
The bacillus licheniformis slant strains is transferred on the slant medium, and cultivation temperature 28-32 ℃, 30 ℃ of optimum culturing temperatures were cultivated about 96-120 hour, covered with the inclined-plane to mycelia and got final product;
1. the liquid first order seed is cultivated: access of above-mentioned slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 28～32 ℃; Rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃.
2. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with the 5-20% inoculum concentration carry out the secondary seed cultivation in 1000 ml shake flasks that 500 milliliters of culture mediums are housed, condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 28～32 ℃; Suitable inoculum concentration is 10%, and rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃.
3. three grades of seed culture of liquid: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle, 1000 milliliters of culture medium loading amounts, were cultivated about 96 hours for 28～32 ℃ by rotary shaking table 80-180 rev/min; Suitable inoculum concentration is 10%, and rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃.
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28～32 ℃ of cultivation temperature, 150 rev/mins of mixing speeds, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 28～32 ℃ of condition of culture cultivation temperature, 150 rev/mins of mixing speeds, incubation time 36-48 hour, be prepared into the bacillus licheniformis bacterial classification.Cultivate and finish bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
Mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 48～72 hours, 80 ℃ of drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
Embodiment 1
The saccharomyces cerevisiae strain preparation: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, and switching goes into to be added with in the fermentation tank of fermentation medium, and the control temperature is 30 ℃, ventilates early stage to cultivating 16 hours, and throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification.Fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
Lactic acid sheet bacterium (Lactobacillus) strain preparation:
1. first order seed is cultivated: lactic acid sheet bacterium slant strains is inserted in 500 ml shake flasks 100 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
2. secondary seed is cultivated: first order seed is inserted 1000 milliliters of secondary seeds according to 10% inoculum concentration shake in the bottle, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle 1000 milliliters of culture medium loading amounts, 100 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa incubation time 36-48 hour, cultivates and finishes bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
The bacillus licheniformis strain preparation:
The bacillus licheniformis slant strains is transferred on the slant medium, and 30 ℃ of cultivation temperature were cultivated 96 hours, covered with the inclined-plane to mycelia and got final product;
1. the liquid first order seed is cultivated: access of above-mentioned slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate condition of culture: 150 rev/mins of rotary shaking tables, cultivated 120 hours for 30 ℃;
2. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 5% inoculum concentration carry out the secondary seed cultivation in 1000 ml shake flasks that 500 milliliters of culture mediums are housed, condition of culture: 150 rev/mins of rotary shaking tables, cultivated 130 hours for 30 ℃;
3. three grades of seed culture of liquid: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle, 1000 milliliters of culture medium loading amounts, 150 rev/mins of rotary shaking tables were cultivated 96 hours for 30 ℃;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28～32 ℃ of cultivation temperature, 150 rev/mins of mixing speeds, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 30 ℃ of condition of culture cultivation temperature, 150 rev/mins of mixing speeds, incubation time 36 hours is prepared into the bacillus licheniformis bacterial classification.Cultivate and finish bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
Mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 48 hours, 80 ℃ of drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
Embodiment 2
The saccharomyces cerevisiae strain preparation: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, and switching goes into to be added with in the fermentation tank of fermentation medium, and the control temperature is 28 ℃, ventilates early stage to cultivating 16 hours, and throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification.Fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
Lactic acid sheet bacterium (Lactobacillus) strain preparation:
1. first order seed is cultivated: lactic acid sheet bacterium slant strains is inserted in 500 ml shake flasks 100 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
2. secondary seed is cultivated: first order seed is inserted 1000 milliliters of secondary seeds according to 10% inoculum concentration shake in the bottle, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle 1000 milliliters of culture medium loading amounts, 100 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa incubation time 36-48 hour, cultivates and finishes bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
The bacillus licheniformis strain preparation:
The bacillus licheniformis slant strains is transferred on the slant medium, and 28 ℃ of cultivation temperature were cultivated 120 hours, covered with the inclined-plane to mycelia and got final product;
1. the liquid first order seed is cultivated: access of above-mentioned slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate condition of culture: 80 rev/mins of rotary shaking tables, cultivated 130 hours for 28 ℃;
2. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 10% inoculum concentration carry out the secondary seed cultivation in 1000 ml shake flasks that 500 milliliters of culture mediums are housed, condition of culture: 80 rev/mins of rotary shaking tables, cultivated 130 hours for 28 ℃;
3. three grades of seed culture of liquid: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle, 1000 milliliters of culture medium loading amounts, 80 rev/mins of rotary shaking tables were cultivated 96 hours for 28 ℃;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 150 rev/mins of mixing speeds, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 28 ℃ of condition of culture cultivation temperature, 150 rev/mins of mixing speeds, incubation time 48 hours is prepared into the bacillus licheniformis bacterial classification.Cultivate and finish bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
Mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 72 hours, 80 ℃ of drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
Embodiment 3
The saccharomyces cerevisiae strain preparation: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, and switching goes into to be added with in the fermentation tank of fermentation medium, and the control temperature is 30 ℃, ventilates early stage to cultivating 16 hours, and throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification.Fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
Lactic acid sheet bacterium (Lactobacillus) strain preparation:
1. first order seed is cultivated: lactic acid sheet bacterium slant strains is inserted in 500 ml shake flasks 100 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
2. secondary seed is cultivated: first order seed is inserted 1000 milliliters of secondary seeds according to 10% inoculum concentration shake in the bottle, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle 1000 milliliters of culture medium loading amounts, 100 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 40 hours is cultivated and is finished bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
The bacillus licheniformis strain preparation:
The bacillus licheniformis slant strains is transferred on the slant medium, and 32 ℃ of cultivation temperature were cultivated 96 hours, covered with the inclined-plane to mycelia and got final product;
1. the liquid first order seed is cultivated: access of above-mentioned slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate condition of culture: 180 rev/mins of rotary shaking tables, cultivated 96 hours for 32 ℃;
2. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 20% inoculum concentration carry out the secondary seed cultivation in 1000 ml shake flasks that 500 milliliters of culture mediums are housed, condition of culture: 180 rev/mins of rotary shaking tables, cultivated 96 hours for 32 ℃;
3. three grades of seed culture of liquid: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle, 1000 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables were cultivated 96 hours for 32 ℃;
4. first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 32 ℃ of cultivation temperature, 150 rev/mins of mixing speeds, incubation time 24 hours;
5. fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 32 ℃ of condition of culture cultivation temperature, 150 rev/mins of mixing speeds, incubation time 36 hours is prepared into the bacillus licheniformis bacterial classification.Cultivate and finish bacteria concentration 5.0 * 10
8Individual/ml.
Culture medium is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
Mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 72 hours, 80 ℃ of drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
Claims (6)
1. biology enzyme additive of protein feed, it is characterized in that, described biology enzyme protein feed is according to brewer's yeast, lactic acid sheet bacterium, bacillus licheniformis different qualities, adopt different condition to cultivate respectively and be prepared into different strain separately, mix the fermentation of dark pond then, be specially, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, behind dress pond, the scraper constant temperature culture certain hour, drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package; Described three kinds of bacterial classifications add toatl proportion in the aniseed culture medium be 5%, and wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.
2. biology enzyme additive of protein feed according to claim 1, it is characterized in that, described saccharomyces cerevisiae process for preparing strain thereof is as follows: slant strains obtains the brewer's yeast seed liquor through the multistage technology that spreads cultivation of routine, switching goes into to be added with in the fermentation tank of fermentation medium, the control temperature is 28～30 ℃, ventilate early stage and cultivated 16 hours, throughput is controlled to be 2.0m
3/ minute, the later stage anaerobism was cultivated 15 hours, was prepared into the saccharomyces cerevisiae bacterial classification; Described fermentation tank culture medium is bean cake powder 10%, wheat bran 15%, brewer's wort 10%, sucrose 1% and corn steep liquor 9%.
3. biology enzyme additive of protein feed according to claim 1 is characterized in that described lactic acid sheet bacterial preparation method is as follows:
⑴ first order seed is cultivated: lactic acid sheet bacterium slant strains is inserted in 500 ml shake flasks 100 milliliters of culture medium loading amounts, 180 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
⑵ secondary seed is cultivated: first order seed is inserted 1000 milliliters of secondary seeds according to 10% inoculum concentration shake in the bottle, condition of culture is identical with first order seed;
⑶ three grades of seed culture: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle 1000 milliliters of culture medium loading amounts, 100 rev/mins of rotary shaking tables, 37 ℃ of cultivation temperature, incubation time 24 hours;
⑷ first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28 ℃ of cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa, incubation time 24 hours;
⑸ fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 37 ℃ of condition of culture cultivation temperature, 100 rev/mins of mixing speeds, ventilation (V/V) 1:0.5, tank pressure 0.05Mpa incubation time 36-48 hour, cultivates and finishes bacteria concentration 5.0 * 10
8Individual/ml;
Described culture medium is bean cake powder 6%, corn flour 1.5%, glucose 2%; Corn steep liquor 1%, PH7.0.
4. biology enzyme additive of protein feed according to claim 1, it is characterized in that, described bacillus licheniformis process for preparing strain thereof is as follows: the bacillus licheniformis slant strains is transferred on the slant medium, cultivation temperature 28-32 ℃, 30 ℃ of optimum culturing temperatures, cultivated about 96-120 hour, and covered with the inclined-plane to mycelia and get final product;
⑴ liquid first order seed is cultivated: access of above-mentioned slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 28～32 ℃; Rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃;
⑵ liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with the 5-20% inoculum concentration carry out the secondary seed cultivation in 1000 ml shake flasks that 500 milliliters of culture mediums are housed, condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 28～32 ℃; Suitable inoculum concentration is 10%, and rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃;
⑶ three grades of seed culture of liquid: secondary seed is inserted 5000 milliliters of three grades of seeds with 10% inoculum concentration shake in the bottle, 1000 milliliters of culture medium loading amounts, were cultivated about 96 hours for 28～32 ℃ by rotary shaking table 80-180 rev/min; Suitable inoculum concentration is 10%, and rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 30 ℃;
⑷ first class seed pot is cultivated: it is the first class seed pot of 150L that three grades of seeds are inserted total measurement (volume) with 10% inoculum concentration, fermentation medium loading amount 100L, 28～32 ℃ of cultivation temperature, 150 rev/mins of mixing speeds, incubation time 24 hours;
⑸ fermented and cultured: it is 1.5 tons of secondary seed jars that the first class seed pot bacterial classification is inserted total measurement (volume) with 10% inoculum concentration, 1 ton of fermentation medium loading amount, 28～32 ℃ of condition of culture cultivation temperature, 150 rev/mins of mixing speeds, incubation time 36-48 hour, be prepared into the bacillus licheniformis bacterial classification.Cultivate and finish bacteria concentration 5.0 * 10
8Individual/ml;
Described culture medium is bean cake powder 6%, corn flour 1.5%, Nacl0.2%, corn steep liquor 1%, pH value 7.0.
5. a biology enzyme additive of protein feed according to claim 1 is characterized in that, mixing dark pond fermentating formula is bean cake powder 40%, corn flour 30%, blood meal 5%, big wing 10%, bacterial classification 5% and cotton dregs 10%; Wherein, each bacterial classification addition in the aniseed culture medium is respectively: saccharomyces cerevisiae bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%; Press manufacturing technique requirent, evenly insert saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification in the modulated aniseed culture medium that makes in proportion, dress pond, 32 ℃ of constant temperature culture of scraper are after 48～72 hours, 80 ℃ of drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package.
6. the preparation method of a biology enzyme additive of protein feed, comprise the steps: that described biology enzyme protein feed is according to brewer's yeast, the lactic acid sheet bacterium, the bacillus licheniformis different qualities, adopt different condition to cultivate respectively and be prepared into different strain separately, mix the fermentation of dark pond then, be specially, in proportion with the saccharomyces cerevisiae bacterial classification, lactic acid sheet bacterial and bacillus licheniformis bacterial classification evenly insert in the modulated aniseed culture medium that makes, the dress pond, behind the scraper constant temperature culture certain hour, drying and crushing, warehouse-in makes described biology enzyme additive of protein feed behind the cooperation test package; Described three kinds of bacterial classifications add toatl proportion in the aniseed culture medium be 5%, and wherein, each bacterial classification addition in the aniseed culture medium is respectively: brewer's yeast bacterial classification 2%, lactic acid sheet bacterial 2%, bacillus licheniformis bacterial classification 1%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105941828A (en) * | 2016-05-16 | 2016-09-21 | 内蒙古工业大学 | Method for producing compound feed from mixed strains through united fermentation |
| CN107494984A (en) * | 2017-09-27 | 2017-12-22 | 沧州旺发生物技术研究所（普通合伙） | A kind of composite microbial feed additive and preparation method and application |
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2013
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105941828A (en) * | 2016-05-16 | 2016-09-21 | 内蒙古工业大学 | Method for producing compound feed from mixed strains through united fermentation |
| CN107494984A (en) * | 2017-09-27 | 2017-12-22 | 沧州旺发生物技术研究所（普通合伙） | A kind of composite microbial feed additive and preparation method and application |
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Application publication date: 20130828 |