CN101112220A - Method for preparing feed additive based on natural plant active component biotransformation - Google Patents
Method for preparing feed additive based on natural plant active component biotransformation Download PDFInfo
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Abstract
The invention relates to a preparation method of a feedstuff additive that biologically transformed from natural bioactive components, and is characterized in preparing feedstuff additive from natural vegetation fermenting technique. The preparation method is that Aspergillus niger and Bacillus subtilis are respectively cultivated, a powder and product can be acquired by fermenting liquid or solid Dandelion with Aspergillus niger or Bacillus subtilis, or by fermenting liquid or solid milk veteh with Bacillus subtilis, and the three dry powders are mixed, sieved, thus acquiring a mixed dry powder end product, i.e., natural vegetation drug feedstuff additive. The invention has the advantages that can improve total contents of flavone and Chlorogenic acid by transforming natural vegetation drug, and decrease the toxic effects and side effects of the natural vegetation. In condition that feedstuff is not added with such harmful additives as antibiotic, quietive and hormone, the adding of the additive of the invention can improve the weight of pig and chicken by 5 to 10 percent, and increase the feedstuff transforming ratio by 5 to 8 percent.
Description
Technical field
The present invention relates to a kind of preparation method of the feed addictive based on the natural plant active component bio-transformation, belong to field of feed additive technology.
Background technology
Though the research and development of natural plant feed additives product have obtained certain progress, and the control to livestock and poultry has obtained effect preferably within the specific limits, but also there are many problem demanding prompt solutions, big as the product addition, active ingredient is not clear, the mechanism of action is unclear, the action effect instability, the research of the single shortage toxicity of formulation secure context, the quality control standard of raw material and product is incomplete etc., cause new natural plants additive research and development dynamics little, the listing new product is less.The existence of these problems makes in the market natural plant feed additives not meet the basic function principle of " trace, efficient " this feed addictive, is difficult to realize industrialization.
Extracted form natural plant technology modernized most important be not as extracting the effective component in the Chinese medicine effectively, in any case then good prescription can not be brought into play therapeutic action.Domestic some enterprise has abandoned the method that falls behind in the traditional Chinese medicine production at present, advanced chemical engineering unit operation and modern analysis means are applied to traditional Chinese medicine extraction, and natural plants is made advanced formulation, add in the feed rationally and effectively, make it to be easy to absorb instant effect.These advanced extraction process technology mainly contain adsorption chromatography technology, continuous countercurrent extraction technology, supercritical extraction technique, Freeze Drying Technique, microencapsulation inclusion technique etc., fermentation technique is used for the fermentation that natural medicinal plants production only is confined to simple process of preparing Chinese medicine processing and some mushroom medicines at present, but how to bring into play the natural plant biological transformation technology still in the research initial stage.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of feed addictive based on the natural plant active component bio-transformation.
For realizing above purpose, technical scheme of the present invention provides a kind of preparation method of the feed addictive based on the natural plant active component bio-transformation, it is characterized in that prepare feed addictive with the natural plants fermentation technique, its method is:
The first step. the preservation of bacterial classification and production
(1) gets aspergillus niger and bacillus subtilis respectively 4 ℃ of following preservations;
(2) aspergillus niger being inoculated in seed enlarges in the culture medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium obtains cultured bacillus subtilis;
(3) bacillus subtilis is inoculated in the slant medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium,, obtain cultured bacillus subtilis bacterial classification;
Second step. aspergillus niger is to the liquid or solid fermentation of dandelion
The dandelion liquid fermentation:
A. the preparation of dandelion water extract: use one ton of extractor, with 1: the ratio adding dandelion of 2-4 and water boil 50 minutes-70 minutes, the dandelion residue is filtered out, use the decompression of the special-purpose decompression of Chinese medicine concentration tank concentrated, then obtain the crude extract of dandelion;
B. aspergillus niger is to the fermentation of dandelion crude extract: with aspergillus niger microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion water extract 55%-65% insert in the aeration tank after evenly mixing, 35-40 ℃ bottom fermentation 2-4 days, obtain pulverous finished product through behind the alcohol precipitation, decompression, drying, pulverizing;
Or to the dandelion solid fermentation:
A. the preparation of dandelion: with micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again;
B. aspergillus niger is to the fermentation of dandelion solid: with aspergillus niger microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion 55%-65% insert in the aeration tank after evenly mixing, 35-40 ℃ of bottom fermentation 4 days-6 days, water is carried, obtain pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing;
The 3rd step. bacillus subtilis is to the liquid or solid fermentation of dandelion
The liquid fermentation of dandelion:
A. the preparation of dandelion water extract: use one ton of extractor, with 1: the ratio adding dandelion of 2-4 and water boil 50 minutes-70 minutes, the dandelion residue is filtered out, decompression concentrates, and then obtains the crude extract of dandelion;
B. bacillus subtilis is to the fermentation of dandelion crude extract: with bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion water extract 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 2-4 days, after alcohol precipitation, decompression, drying, pulverizing, obtain pulverous finished product;
Or to the dandelion solid fermentation:
A. the preparation of dandelion: with micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again;
B. bacillus subtilis is to the fermentation of dandelion solid: with bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 4-6 days, through water carry, alcohol precipitation, decompression, drying, pulverizing obtain pulverous finished product;
The 4th step. bacillus subtilis is to the liquid or solid fermentation of the Radix Astragali
The liquid fermentation of the Radix Astragali:
A. the preparation of water extraction of astragalus membranaceus: use one ton of extractor, with 1: the ratio adding Radix Astragali of 2-4 and water boil 50 minutes-70 minutes, Radix Astragali residue is filtered out, decompression concentrates, and then obtains the crude extract of the Radix Astragali;
B. bacillus subtilis is to the fermentation of Radix Astragali crude extract
With bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and water extraction of astragalus membranaceus 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 2-4 days, after alcohol precipitation, decompression, drying, pulverizing, obtain pulverous finished product;
Or to Radix Astragali solid fermentation:
A. the preparation of the Radix Astragali
With micronizer astragalus membranaceus powder is broken to 80 orders, with 35% water logging bubble, standby again;
B. bacillus subtilis is to Radix Astragali solid fermentation
With bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and Radix Astragali 55%-65% insert in the aeration tank after evenly mixing, 35 ℃ of-40 ℃ of bottom fermentations 4 days-6 days are carried, are obtained pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing through water;
The 5th step. preparation natural medicinal plants feed addictive
Dry powder 25%-35%, the dry powder 25%-35% of step 3, dry powder 25%-35%, the licorice powder 5%-15% of step 4 of step 2 are mixed, cross 80 mesh sieves, get mixed powder finished product, i.e. this feed addictive finished product.
It is sucrose 2%-4% that described bacillus subtilis seed is optimized fermentation medium, peptone 0.5%-1.5%, yeast extract 0.3%-0.6%, potassium dihydrogen phosphate (KH
2PO
4) 0.3%-0.6%, ammonium sulfate ((NH
4)
2SO
4) 0.2%-0.4%, calcium carbonate (CaCO
3) 0.1%-0.3%, distilled water 93%-95%, the pH value is 6.5-7.5.
It is glucose 5%-7% that the seed of described aspergillus niger is optimized fermentation medium, soya-bean cake 0.5%-1.5%, and ammonium sulfate 0.4%-0.6%, calcium chloride 0.2%-0.4%, distilled water 91.5%-93.5%, pH value are 4～6.5.
Described carbon source is glucose, amion acetic acid, sucrose, corn flour etc.; Described nitrogenous source is urea, ammonium sulfate, peptone etc.; Described inorganic salts are calcium carbonate, potassium dihydrogen phosphate, magnesium sulfate.
The present invention utilizes the natural plants fermentation technique to prepare feed addictive, multiple abilities such as microorganism has oxidation, esterification, methylates, hydroxylating, reducing, the natural plants fermentation is after bio-transformation, its active ingredient can fully be separated, be extracted, make it have more biologically active, and contain a large amount of organized enzymes, can be absorbed rapidly by the body tissue cell after taking, reach and get rid of illness, be healthy and strong, strengthen immunoregulatory function, bring into play the drug action of this natural drug of natural plants better, thereby the reduction drug cost improves effect.
Advantage of the present invention is:
1. the present invention does not add in mixed feed under harmful additive prerequisites such as antibiotic, sedative and hormone, adds the present invention, and pig, chicken weightening finish improve about 5-10%, and feed conversion rate improves 5-8%;
2. fermentation provides new technology for exploring, improve the natural medicinal plants pharmaceutical technology
Be culture medium with the natural medicinal plants or in culture medium, add an amount of natural medicinal plants, can transform the back to natural medicinal plants and improve flavones, chlorogenic acid contents, reduce the toxic and side effect of natural medicinal plants or improve its drug effect, its principle is a decomposition and inversion ability of utilizing enzyme powerful, not only can be to the fiber of Chinese medicine in the culture medium, material such as carbohydrate and albumen is used, and in metabolic process, can some compositions in the natural medicinal plants be transformed, thereby raising flavones, the ratio of chlorogenic acid and tiring, composition in the natural medicinal plants also can promote the growth of medicinal fungi or the generation of activated product, can produce like this comprise the various active composition and drug effect simpler, the novel fermentation preparation that the prescription compatibility is better;
3. the extraction and the analysis of pharmaceutically active ingredient during fermentation technique helps
Fermentation technique is applied to the extraction of natural medicinal plants, the natural medicinal plants recovery rate is improved, active ingredient obtains purifying, but also can do some structure of modification to Effective Components of Chinese Herb by adopting specific enzymes such as some hydroxylases, because what growth of microorganism mainly consumed is conventional substances such as vegeto-animal protein, sugar, and active ingredient is had inspissation;
4. fermentation technique helps improving natural medicinal plants pharmacology and toxicological effect
Natural medicinal plants with fermentation method production is compared with unfermentable natural medicinal plants, and change has taken place in its active ingredient effect at aspects such as pharmacology, toxicitys, through the decomposition of microorganism original non-digestible material is become and is easy to absorb;
5. fermentation technique is used for the natural medicinal plants process of preparing Chinese medicine
Modern Fermentation Engineering confirms that microorganism has esterification; oxidation; acetylation; reducing; methylate; multiple ability such as hydroxylating and carbonylation; therefore utilize microorganism to ferment and concoct the bioconversion reaction that natural medicinal plants comes down to the modification of a kind of enzymatic natural medicinal plants active ingredient; microorganism is owing to self existing abundant enzyme system that the ability of decomposition and inversion material under temperate condition is arranged; ferment altogether by microorganism and natural medicinal plants and to carry out natural medicinal plants and concoct and to change the property of medicine by a larger margin than general physical chemical means; improve drug effect, reduce toxic and side effect, enlarge indication.
The specific embodiment
The invention will be further described below in conjunction with embodiment.
The present invention adopts aspergillus niger and bacillus subtilis to be provided by Ministry of Agriculture bacterial classification preservation center.
Embodiment 1: by different microorganisms dandelion, Radix Astragali crude extract are carried out bio-transformation and produce feed addictive.
Get aspergillus niger and bacillus subtilis respectively 4 ℃ of following preservations;
Step 1: aspergillus niger is to the bio-transformation of dandelion crude extract
This example mainly is to improve dandelion active ingredient content of total flavone, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of aspergillus niger seed and fermentation medium
Aspergillus niger is inoculated in seed to be enlarged in the culture medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium obtains cultured bacillus subtilis;
It is seed culture medium that the seed of aspergillus niger enlarges culture medium: corn flour 3%, wheat bran 2%, ammonium sulfate 0.15%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.05%, distilled water 94.6%, pH are 6.0.
It is glucose (sucrose) 6%, soya-bean cake (yeast extract or urea) 1%, ammonium sulfate 0.5%, calcium chloride 0.3%, distilled water 92.2% that the seed of aspergillus niger is optimized fermentation medium, and PH is 4～6.5.
(2) preparation of dandelion water extract
Get one ton of extractor, then take by weighing 200 kilograms of dandelions, add 500 kg water and boiled 60 minutes, the dandelion residue is filtered out, decompression concentrates, and then obtains the crude extract of dandelion;
(3) aspergillus niger is to the fermentation of dandelion crude extract
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the dandelion water extract 60% of 2: 1: 1 ratios evenly mix with aspergillus niger microbial inoculum 20%, 40 ℃ of bottom fermentations 3 days obtain pulverous finished product after alcohol precipitation, decompression, drying, pulverizing;
Step 2: bacillus subtilis is to the bio-transformation of dandelion crude extract
This example mainly is the content that improves dandelion active ingredient total chlorogenic acid, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of bacillus subtilis seed and fermentation medium
Bacillus subtilis is inoculated in the slant medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium obtains cultured bacillus subtilis bacterial classification;
Bacillus subtilis slant medium composition is beef extract 0.5g, peptone 1.0g, sodium chloride 0.5g, water 100ml, agar 2g, and the pH nature is in 0.1mpa steam sterilizing 30min.
The bacillus subtilis seed is optimized fermentation medium: sucrose 3%, peptone 1%, yeast extract 0.5%, potassium dihydrogen phosphate (KH
2PO
4) 0.45%, ammonium sulfate ((NH
4)
2SO
4) 0.25%, calcium carbonate 0.2%, distilled water 94.6%, pH7.0.
(2) preparation of dandelion water extract
Get one ton of extractor, then take by weighing 200 kilograms of dandelions, added the 500l water boil 60 minutes, the dandelion residue is filtered out, decompression concentrates, and then obtains the crude extract of dandelion.
(3) bacillus subtilis is to the fermentation of dandelion crude extract
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the dandelion water extract 60% of 2: 1: 1 ratios evenly mix with bacillus subtilis microbial inoculum 20%, 35-40 ℃ of bottom fermentation 3 days, alcohol precipitation, decompression, drying obtains pulverous finished product after the pulverizing.
Step 3: bacillus subtilis is to the bio-transformation of Radix Astragali crude extract
This example mainly is to improve Radix Astragali active ingredient content of total flavone, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of water extraction of astragalus membranaceus
Get one ton of extractor, then take by weighing 200 kilograms of Radixs Astragali, added the 500l water boil 60 minutes, the dandelion residue is filtered out, decompression concentrates, and then obtains the crude extract of the Radix Astragali.
(2) bacillus subtilis is to the fermentation of Radix Astragali crude extract
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the water extraction of astragalus membranaceus 60% of 2: 1: 1 ratios evenly mix with bacillus subtilis microbial inoculum 20%, 35-40 ℃ of bottom fermentation 3 days, alcohol precipitation, decompression, drying obtains pulverous finished product after the pulverizing.
Step 4: bio-transformation is produced the feed addictive finished product and is made
Dry powder 30%, the dry powder 30% of step 2, the dry powder 30% of step 3, the licorice powder 10% of step 1 are mixed, cross 80 mesh sieves, get mixed powder finished product, i.e. this feed addictive finished product.
In use, evenly mixing and stirring the back according to different livestock and poultry in different proportion and feed feeds: the fowl poultry kind animal is in 3 ‰ ratios; The aquatic product animal is in 2 ‰ ratios.
When being used for feed granulating, this product granulation tailored version being added in feed or the premix in the ratio of 1-3 ‰ granulate.
Embodiment 2: by different microorganisms dandelion, Radix Astragali solid fermentation are produced feed addictive
Step 1: aspergillus niger is to the dandelion solid fermentation
This example mainly is to improve dandelion active ingredient content of total flavone, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of aspergillus niger seed and seed optimization fermentation medium is identical with embodiment 1;
(2) preparation of dandelion
With micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again.
(3) aspergillus niger is to the fermentation of dandelion solid
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the dandelion 60% of 2: 1: 1 ratios evenly mix with aspergillus niger microbial inoculum 20%, 35-40 ℃ of bottom fermentation 5 days, water is carried, alcohol precipitation, decompression, drying is pulverized, and obtains pulverous finished product.
Step 2: bacillus subtilis is to the dandelion solid fermentation
This example mainly is the content that improves dandelion active ingredient total chlorogenic acid, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of bacillus subtilis seed and seed optimization fermentation medium is identical with embodiment 1;
(2) preparation of dandelion
With micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again.
(3) bacillus subtilis is to the fermentation of dandelion crude extract
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the dandelion 60% of 2: 1: 1 ratios evenly mix with bacillus subtilis microbial inoculum 20%, 35-40 ℃ of bottom fermentation 5 days, water is carried, obtain pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing.
Step 3: bacillus subtilis is to Radix Astragali solid fermentation
This example mainly is to improve Radix Astragali active ingredient content of total flavone, to improve the immunity of livestock and poultry, promotes growth of animal.
(1) preparation of the Radix Astragali
With micronizer astragalus membranaceus powder is broken to 80 orders, with 35% water logging bubble, standby again.
(2) bacillus subtilis is to Radix Astragali solid fermentation
Insert in the aeration tank after in addition amion acetic acid, urea, inorganic salts 20% and the Radix Astragali 60% of 2: 1: 1 ratios evenly mix with bacillus subtilis microbial inoculum 20%, 35-40 ℃ of bottom fermentation 5 days, water is carried, obtain pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing.
Step 4: bio-transformation is produced the feed addictive finished product and is made
Dry powder 30%, the dry powder 30% of step 2, the dry powder 30% of step 3, the licorice powder 10% of step 1 are mixed together, cross 80 mesh sieves, get mixed powder finished product, i.e. this feed addictive finished product.
In use, evenly mixing and stirring the back according to different livestock and poultry in different proportion and feed feeds: the fowl poultry kind animal is in 3 ‰ ratios; The aquatic product animal is in 2 ‰ ratios.
When being used for feed granulating, this product granulation tailored version being added in feed or the premix in the ratio of 1-3 ‰ granulate.
Index determining
The preparation of need testing solution and the mensuration of general flavone content:
A. the preparation of reference substance solution: precision takes by weighing the control substance of Rutin 10mg that is dried to constant weight at 120 ℃, puts in the 100mL volumetric flask, and the accurate 10mL of absorption 0.85 ethanol dissolves, and adding distil water is diluted to scale (1mL=0.1mg rutin).
B. the preparation of need testing solution: precision takes by weighing this product 40g, adds 1000mL water, boils after the Radix Astragali becomes white by yellow, adds 70% ethanol, and rotary evaporation 30min obtains the Chinese medicine concentrate, and is standby.
C. the calibration curve precision is measured rutin titer 0.0,0.5,1.0,2.0,3.0,4.0, and 5.0mL puts respectively in the 10mL volumetric flask, adds 5% sodium nitrite solution 0.4mL in order respectively and shakes up, and places 6min.Add 10% liquor alumini chloridi 0.4mL, shake up, place 6min.Add 4% sodium hydroxide solution 4mL again, and adding distil water is diluted to scale, shakes up, place 15min.Titer with 0.0mL in the standard liquid is cooked blank, measures each absorbance in wavelength 510nm place, and the drawing standard curve gets regression equation.
The preparation of need testing solution and the mensuration of chlorogenic acid content:
A. the preparation of reference substance solution: precision takes by weighing the chlorogenic acid reference substance 10mg that is dried to constant weight at 120 ℃, puts in the 100mL volumetric flask, and the accurate 10mL of absorption 0.85 ethanol dissolves, and adding distil water is diluted to scale (1mL=0.1mg chlorogenic acid).
B. the drafting of chlorogenic acid calibration curve: according to the chlorogenic acid standard liquid of the concentration known tonsure that is disposed, with ferric trichloride special chromogenic reaction takes place at 35 ℃, under the 755nm condition, be in charge of after the photometric determination absorbance drawing standard curve with visible light.
C. the preparation of need testing solution: this product of 10g is added in the entry, boil 30min, and it is additional constantly to add water, and constant volume is surveyed volume.Get the extract of dandelion, behind the alcohol precipitation, get 8ml liquid, in 35 ℃ water-bath, add the FeCl of 0.8ml
3Solution after concussion shakes up, leaves standstill 60min, is to detect wavelength with 755nm, measures the absorbance of extract with visible spectrophotometer.Obtain chlorogenic acid content in this product according to the chlorogenic acid calibration curve.
The contrast table of feed addictive parameter
Table 1 distinct methods prepares the parameter of feed addictive chlorogenic acid and total brass
| The preparation method | Chlorogenic acid (%) | General flavone (%) |
| Traditional extraction process | 1.5 | 4.0 |
| The solid fermentation method for post extraction | 2.3 | 6.5 |
| The liquid fermentation extracting method | 2.9 | 7.8 |
Claims (6)
1. the preparation method based on the feed addictive of natural plant active component bio-transformation is characterized in that, prepares feed addictive with the natural plants fermentation technique, and its method is:
The first step. the preservation of bacterial classification and production
(1) gets aspergillus niger and bacillus subtilis respectively 4 ℃ of following preservations;
(2) aspergillus niger being inoculated in seed enlarges in the culture medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium obtains cultured bacillus subtilis;
(3) bacillus subtilis is inoculated in the slant medium, putting into 35 ℃-40 ℃ constant incubator cultivated 2-3 days, inserting seed again optimizes in the constant temperature shaking table incubator that fermentation medium is placed on 35 ℃-40 ℃ and cultivated 2-3 days, every 300ml triangular flask loading amount 50ml fluid nutrient medium obtains cultured bacillus subtilis bacterial classification;
Second step. aspergillus niger is to the liquid or solid fermentation of dandelion
The dandelion liquid fermentation:
A. the preparation of dandelion water extract: use one ton of extractor, with 1: the ratio adding dandelion of 2-4 and water boil 50 minutes-70 minutes, the dandelion residue is filtered out, use the decompression of the special-purpose decompression of Chinese medicine concentration tank concentrated, then obtain the crude extract of dandelion;
B. aspergillus niger is to the fermentation of dandelion crude extract: with aspergillus niger microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion water extract 55%-65% insert in the aeration tank after evenly mixing, 35-40 ℃ bottom fermentation 2-4 days, obtain pulverous finished product through behind the alcohol precipitation, decompression, drying, pulverizing;
Or to the dandelion solid fermentation:
A. the preparation of dandelion: with micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again;
B. aspergillus niger is to the fermentation of dandelion solid: with aspergillus niger microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion 55%-65% insert in the aeration tank after evenly mixing, 35-40 ℃ of bottom fermentation 4 days-6 days, water is carried, obtain pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing;
The 3rd step. bacillus subtilis is to the liquid or solid fermentation of dandelion
The liquid fermentation of dandelion:
A. the preparation of dandelion water extract: use one ton of extractor, with 1: the ratio adding dandelion of 2-4 and water boil 50 minutes-70 minutes, the dandelion residue is filtered out, decompression concentrates, and then obtains the crude extract of dandelion;
B. bacillus subtilis is to the fermentation of dandelion crude extract: with bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion water extract 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 2-4 days, after alcohol precipitation, decompression, drying, pulverizing, obtain pulverous finished product;
Or to the dandelion solid fermentation:
A. the preparation of dandelion: with micronizer dandelion is crushed to 80 orders, with 35% water logging bubble, standby again;
B. bacillus subtilis is to the fermentation of dandelion solid: with bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and dandelion 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 4-6 days, through water carry, alcohol precipitation, decompression, drying, pulverizing obtain pulverous finished product;
The 4th step. bacillus subtilis is to the liquid or solid fermentation of the Radix Astragali
The liquid fermentation of the Radix Astragali:
A. the preparation of water extraction of astragalus membranaceus: use one ton of extractor, with 1: the ratio adding Radix Astragali of 2-4 and water boil 50 minutes-70 minutes, Radix Astragali residue is filtered out, decompression concentrates, and then obtains the crude extract of the Radix Astragali;
B. bacillus subtilis is to the fermentation of Radix Astragali crude extract
With bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and water extraction of astragalus membranaceus 55%-65% insert in the aeration tank after evenly mixing, 35 ℃-40 ℃ bottom fermentation 2-4 days, after alcohol precipitation, decompression, drying, pulverizing, obtain pulverous finished product;
Or to Radix Astragali solid fermentation:
A. the preparation of the Radix Astragali
With micronizer astragalus membranaceus powder is broken to 80 orders, with 35% water logging bubble, standby again;
B. bacillus subtilis is to Radix Astragali solid fermentation
With bacillus subtilis microbial inoculum 15%-25% in addition 2: 0.5-1.5: the carbon source of 0.5-1.5 ratio, nitrogenous source, inorganic salts 15%-25% and Radix Astragali 55%-65% insert in the aeration tank after evenly mixing, 35 ℃ of-40 ℃ of bottom fermentations 4 days-6 days are carried, are obtained pulverous finished product behind the alcohol precipitation, decompression, drying, pulverizing through water;
The 5th step. preparation natural medicinal plants feed addictive
Dry powder 25%-35%, the dry powder 25%-35% of step 3, dry powder 25%-35%, the licorice powder 5%-15% of step 4 of step 2 are mixed, cross 80 mesh sieves, get mixed powder finished product, i.e. this feed addictive finished product.
2. the preparation method of a kind of feed addictive based on the natural plant active component bio-transformation according to claim 1, it is characterized in that, it is sucrose 2%-4% that described bacillus subtilis seed is optimized fermentation medium, peptone 0.5%-1.5%, yeast extract 0.3%-0.6%, potassium dihydrogen phosphate (KH
2PO
4) 0.45%, ammonium sulfate ((NH
4)
2SO
4) 0.25%, calcium carbonate 0.2%, distilled water 93%-95%, the pH value is 6.5-7.5.
3. the preparation method of a kind of feed addictive based on the natural plant active component bio-transformation according to claim 1, it is characterized in that, it is glucose 5%-7% that the seed of described aspergillus niger is optimized fermentation medium, soya-bean cake 0.5%-1.5%, ammonium sulfate 0.4%-0.6%, calcium chloride 0.2%-0.4%, distilled water 91.5%-93.5%, pH value are 4～6.5.
4. the preparation method of a kind of feed addictive based on the natural plant active component bio-transformation according to claim 1 is characterized in that described carbon source is amion acetic acid, glucose, sucrose etc.
5. the preparation method of a kind of feed addictive based on the natural plant active component bio-transformation according to claim 1 is characterized in that described nitrogenous source is urea, peptone etc.
6. the preparation method of a kind of feed addictive based on the natural plant active component bio-transformation according to claim 1 is characterized in that described inorganic salts are calcium chloride, magnesium sulfate etc.
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