CN1648237A - Process for preparing high active powder composite green storage inoculating agent and its carrier adsorption agent - Google Patents
Process for preparing high active powder composite green storage inoculating agent and its carrier adsorption agent Download PDFInfo
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- CN1648237A CN1648237A CNA2004100658084A CN200410065808A CN1648237A CN 1648237 A CN1648237 A CN 1648237A CN A2004100658084 A CNA2004100658084 A CN A2004100658084A CN 200410065808 A CN200410065808 A CN 200410065808A CN 1648237 A CN1648237 A CN 1648237A
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- cellulase
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Abstract
The present invention belongs to the field of silage fermenting promoter technology. The present invention includes simultaneous inoculation of and asynchronous with of lactostreptococcus as initial fermenting bacteria and lactobacillus as subsequent fermenting bacteria, normal temperature carrier adsorption post-processing technology, optimizing the recipes of seed culture medium activating factor, industrial fermenting culture medium and protecting agent, and re-compounding fermented lactic acid bacteria and cellulase. The present invention has the features of mild production condition, low cost, high product activity, etc. The product may fit different silage materials and different silage conditions and can promote the fermentation of lactic acid bacteria and the homotypic fermentation, control negative fermentation, reduce loss of nutrients during fermentation and make full use of resource.
Description
Technical field
The present invention relates to the preparation method of a kind of high active powder composite green storage inoculating agent and carrier thereof absorption, belong to silage additive technical field in the fodder industry, classification more specifically belongs to silage fermentation promotor.
Technical background
China's year per capita the situation of 400 kilograms of grains can not change in the long term, that is China cannot take out more grain as feed grain.According to estimates, by 2000, the breach of annual feed grain will reach 4,000 ten thousand tons.Utilize feedstock production silages such as agricultural crop straw, herbage, aquatic feed, the exploitation feed resource improves efficiency of feed utilization, and the herbvore domestic animal of developing grain-saving type is improved meat product output, as China's fundamental state policy greater and fasterly.
China is large agricultural country, and the various green and rough feedss of annual production are about more than 500,000,000 tons, wherein has only about 30% as the cattle and sheep feed, and all the other major parts are burnt or thrown away, and cause the significant wastage of potential feed resource.The benefit of ensiling is that the nutritive substance loss is few, and the dry-matter loss is no more than 10% during ensiling, and protein and carotene loss are seldom.And dry-matter and leaf protein loss can reach 20-30% during hay curing.Ensiling is not subjected to weather restriction, can concentrate making, can guarantee that the long-term four seasons of green fodder supply.Use microbe additive can improve the quality of ensilage in the ensilage, promote the silage fermentation process, strengthen homofermentation, reduce the nitrogen nutrition loss, reduce voltaile fatty acid and generate, improve palatability, reduce toxic side effect, prevent corrupt histamine and the mycotoxin that produces.Meta-bolites bacteriocin of milk-acid bacteria and whole cell peptidoglycan have excellent antibiotic and bacteriostatic action in addition.Traditional ensiling technology nutritive substance in ensilage loses big, opens easily corruption of back, cellar for storing things.Utilize the microbial preparation that is specifically designed to ensiling of modern biotechnology development, can overcome the problems referred to above effectively, obtained the extensive approval of academia and herding manufacturing enterprise, and in the North America and Europe obtained applying widely.
In ensilage, artificially add microbial preparation (comprising viable bacteria body, enzyme), this is the most active field of nearly section time.The U.S., Germany, Britain drop into the work that conducts a research of a large amount of human and material resources, financial resources one after another, and have released some products successively.Its technical essential is: promote propagation and the homofermentation of milk-acid bacteria, reduce the pH value fast to optimum range; Reduce the consumption of nutritive substance in the ensilage as much as possible; Improve the utilization ratio of silage nutrient, thereby improved the quality of ensilage.Technological difficulties are: the high yield lactobacillus strain; The suitable proportion of various bacterial strains and plurality of enzymes; Promote the activator of bacterial strain, enzyme reaction; The protection of product and post-treatment technology.
The domestic patent of still not having at present this respect, the patent of the U.S. different green storage inoculating agent aspect is more: as U.S.Pat No.4842871,4981705,5747020,6054148 etc.
Summary of the invention
The preparation method who the purpose of this invention is to provide the absorption of a kind of high active powder composite green storage inoculating agent and carrier thereof.Can improve ensiling straw lactic acid content more than 500% after adopting green storage inoculating agent of the present invention, reduce more than the pH0.5, it is about 10% to reduce acid detergent fiber, and it is about 50% that amino nitrogen reduces, and has improved the quality of ensilage significantly.
Technical scheme of the present invention: adopting streptococcus uberis (Streptococcus lactis CVCC590) is initial fermented bacterium; (Lactococcus lactis AS1.557 or AS1.9) is follow-up fermented bacterium with lactobacillus; with tunning add protective material and and cellulase compounding; be prepared into powder by absorption of normal temperature carrier and post-treatment technology, realize the long-term preservation of product.Milk-acid bacteria content is not less than 3 * 10 in the product
10Cfu/g, cellulase activity are not less than 5000 units/g, and the activity of milk-acid bacteria survival rate and cellulase is greater than 80% in 1 year product of stored refrigerated.The application of this product is evenly to make an addition in the silage with 0.5-0.6 ‰ mass ratio, can improve lactic acid content more than 500% after ensiling is added, and reduces more than the pH0.5, and it is about 10% to reduce neutral detergent fiber, and amino nitrogen reduces about 50%.
The preparation method of product of the present invention:
(1) bacterial classification
With streptococcus uberis as initial fermented bacterium, with lactobacillus as follow-up fermented bacterium;
(2) zymotechnique
The composition of seed culture medium
With MRS substratum (peptone 10g, meat extract 10g, yeast extract paste 5g, K
2HPO
42g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, tween 80 1ml, MgSO
47H
2O 0.58g, MnSO
44H
2O 0.25g, above component is settled to 1 liter with distilled water) be the basis, quality percentage meter in addition in addition: tomato juice 5-20%, Radix Dauci Sativae juice 1-15%, aminothiopropionic acid hydrochloride 0.02-1.0%, accent pH are 6.0-6.8,115 ℃ of steam sterilizings 20 minutes.
The suitability for industrialized production fermention medium is formed
In the quality percentage: rice glucose syrup 3-5% or maize dextrose syrup 3-8%, yeast 0.5-5%, all the other quality percentage compositions are water, and mineral substance is formed (1 liter content in substratum) K
2HPO
41-5g, dibasic ammonium citrate 1-6g, sodium acetate 2-8g, MgSO
47H
2O 0.3-1g, MnSO
44H
2O 0.1-1g.
Culture condition
Seed culture: adopt inoculation, asynchronous fermentation technique simultaneously, streptococcus uberis and lactobacillus are inserted 10 liters of seeding tanks simultaneously cultivate, adopt seed culture medium, each bacterial classification inoculation amount is 4-8%, and 35-40 ℃ of incubator cultivated 18 hours.
Industrial fermentation is produced: seed culture fluid is inoculated in the fermention medium by the 4-8% inoculum size, in 150 liters of fermentor tanks, carry out liquid fermenting, temperature 35-40 ℃, ferment tank pressure 0.02-0.04Mpa, nitrogen flow 0.12-0.15Mpa, stirring velocity 30-120rpm, initial pH6.0-6.5, fermentation time 16-23 hour.
(3) the composite green storage inoculating agent is composite
Centrifugal bacterium mud and the protective material that obtains of compound lactobacillus fermented liquid that suitability for industrialized production is obtained mixes; adding cellulase then continues to be mixed into evenly; the absorption carrier gac and the abundant mixing of attapulgite that add dry sterilization are at last crossed the 18-25 mesh sieve and are obtained the good powder of free-running property.
Bacterium mud and protectant mass volume ratio are 1: 1-4 (W/V), and 1 kilogram of bacterium mud adds the cellulase of 0.5-1 kilogram, and 1 kilogram of bacterium mud need add 0.3-1.5 kilogram gac and 0.5-1.2 kilogram attapulgite.
Protectant quality percentage prescription consists of: the 5-20% skim-milk, 1-8% trehalose, 5-25% glycerine, all the other are water.
(4) keep-alive post-treatment: the powder that makes is sealed packing with aluminium foil bag,, or vacuumize the room temperature preservation 0-4 ℃ of stored refrigerated.
Beneficial effect of the present invention: these product development and application can adapt to the needs of different ensiling raw materials and ensiling condition, promote the fermentation of the milk-acid bacteria in the ensilage effectively, promote homofermentation, control bad fermentation, reduce the loss of nutrient substance in the ensilage, improve the quality of ensilage, improve the agreeable to the taste of ensilage, guarantee the ensiling success.
The Application and Development of additives for ensiling, can realize utilizing feedstock production silages such as crop stalk, herbage and aquatic feed, expand feed resource, alleviate the grain feed inadequate resource, the livestock industry of development joint grain, the output of raising livestock product, improve the livestock product quality, help the adjustment of the structure of rural undertaking, increase the income of aquaculture, improve the export competitiveness of China's livestock product.
The present invention adopts normal temperature carrier absorption prepare the method for high active powder composite green storage inoculating agent, has working condition gentleness, simple, with low cost, the active high outstanding advantage of product of technology, the suitability for industrialized production that helps product with apply.
Embodiment
Embodiment 1.
The preparation of high active powder composite green storage inoculating agent
It is initial fermented bacterium that fermented bacterium adopts streptococcus uberis CVCC590, and plant lactobacillus AS1.557 is follow-up fermented bacterium.
After seed culture, seed fermentation liquid is inoculated in the suitability for industrialized production fermentor tank, its substratum consists of: yeast 0.5-5%, the rice glucose syrup of 3-5%, mineral substance form (1 liter content in substratum) K
2HPO
41-5g, dibasic ammonium citrate 1-6g, sodium acetate 2-8g, MgSO
47H
2O 0.3-1g, MnSO
44H
2O 0.1-1g.
Culture condition
Seed culture: adopt inoculation, asynchronous fermentation technique simultaneously, streptococcus uberis and plant lactobacillus are inserted 10 liters of seeding tanks simultaneously cultivate, adopt seed culture medium, each bacterial classification inoculation amount is 4% inoculum size, and 40 ℃ of incubators were cultivated 18 hours;
Suitability for industrialized production: 4% inoculum size, 37 ℃, ferment tank pressure 0.02-0.04Mpa, nitrogen flow 0.12-0.15Mpa, stirring velocity 30-120rpm, initial pH6.0-6.5, fermentation time 23 hours;
(3) preparation of composite green storage inoculating agent
Centrifugal bacterium mud and the protective material that obtains of compound lactobacillus fermented liquid that suitability for industrialized production is obtained mixes; adding cellulase then continues to be mixed into evenly; the abundant mixing of absorption carrier gac, attapulgite that adds dry sterilization is at last crossed the 18-25 mesh sieve and is obtained the good powder of free-running property.
Bacterium mud and protectant ratio are 1: 1 (W/V), and 1 kilogram of bacterium mud adds 0.5 kilogram cellulase, and 1 kilogram of bacterium mud need add 0.3 kilogram of gac and 1.2 kilograms of attapulgites.
Protectant prescription consists of: 20% skim-milk, and 1% trehalose, 12% glycerine, all the other are water.
(4) keep-alive post-treatment: the powder that makes is sealed packing with aluminium foil bag,, or vacuumize the room temperature preservation 0-4 ℃ of stored refrigerated.
Embodiment 2
The preparation of high active powder composite green storage inoculating agent
It is initial fermented bacterium that fermented bacterium adopts streptococcus uberis CVCC590, and lactobacillus lactis AS1.9 is follow-up fermented bacterium.
After seed culture, seed fermentation liquid is inoculated in the suitability for industrialized production fermentor tank, its substratum consists of: yeast 0.5-5%, the maize dextrose syrup of 3-8%, all the other quality percentage components are water, mineral substance is formed (1 liter content in substratum) K
2HPO
41-5g, dibasic ammonium citrate 1-6g, sodium acetate 2-8g, MgSO
47H
2O 0.3-1g, MnSO
44H
2O 0.1-1g.
Culture condition
Seed culture: adopt inoculation, asynchronous fermentation technique simultaneously, streptococcus uberis and lactobacillus are inserted 10 liters of seeding tanks simultaneously cultivate, adopt seed culture medium, each bacterial classification inoculation amount is 8% inoculum size, and 35 ℃ of incubators were cultivated 18 hours.
Suitability for industrialized production: 8% inoculum size, 37 ℃, ferment tank pressure 0.02-0.04Mpa, nitrogen flow 0.12-0.15Mpa, stirring velocity 30-120rpm, initial pH6.0-6.5, fermentation time 16 hours.
(3) preparation of composite green storage inoculating agent
Centrifugal bacterium mud and the protective material that obtains of compound lactobacillus fermented liquid that suitability for industrialized production is obtained mixes; adding cellulase then continues to be mixed into evenly; the abundant mixing of absorption carrier gac, attapulgite that adds dry sterilization is at last crossed the 18-25 mesh sieve and is obtained the good powder of free-running property.
Bacterium mud and protectant ratio are 1: 4 (W/V), and 1 kilogram of bacterium mud adds 1 kilogram cellulase, and 1 kilogram of bacterium mud need add 1.5 kilograms of gacs and 0.5 kilogram of attapulgite.
The prescription of protection dosage consists of: 5% skim-milk, and 8% trehalose, 20% glycerine, all the other are water.
(4) keep-alive post-treatment: the powder that makes is sealed packing with aluminium foil bag,, or vacuumize the room temperature preservation 0-4 ℃ of stored refrigerated.
Embodiment 3.
Above-mentioned substratum with this specification sheets is inoculated into seed culture medium with streptococcus uberis and lactobacillus, prepares seed in 10 liters of fermentor tanks, inserts 150 liters of ferment tanks then.Fermented liquid is obtained bacterium mud through the continuous high speed centrifugation, and the method by this specification sheets adds protective material, cellulase and carrier sorbent material then, product preparation is become the powder of 18-25 mesh sieve.With the aluminium foil coating-dividing sealing 0-4 ℃ of preservation.
The green storage inoculating agent that to produce with this method is directly evenly spread in the silage corn straw by the addition of 0.5 kg/ton.
The chemical composition analysis of 60 days ensiling corns of table 1 (g/kg dry-matter)
Project control group test group
Dry-matter 215.5 220.7
Soluble-carbohydrate 40.19 34.00
Neutral detergent fiber 667.8 630.7
Acid detergent fiber 447.0 412.3
Crude protein 92.7 109.0
pH 4.32 3.60
Lactic acid 6.78 52.85
Acetate (* 10
-3) 14.4 2.64
Propionic acid (* 10
-4) 2.50 5.15
Butyric acid (* 10
-5) 2.07 2.70
NH
3-N 0.321 0
Embodiment 4.
Above-mentioned substratum with this specification sheets is inoculated into seed culture medium with streptococcus uberis and lactobacillus, prepares seed in 10 liters of fermentor tanks, inserts 150 liters of ferment tanks then.Method by this specification sheets obtains bacterium mud with fermented liquid through the continuous high speed centrifugation, adds protective material, cellulase and carrier sorbent material, product preparation is become the powder of 18-25 mesh sieve.Vacuumize normal temperature with the aluminium foil packing and preserve preservation.
The green storage inoculating agent that to produce with this method is directly evenly spread in the ensiling rye grass by the addition of 0.6 kg/ton.
The chemical composition analysis of 40 days ensiling rye grasses of table 2 (g/kg dry-matter)
Project control group test group
Dry-matter 207.15 215.25
pH 4.98 3.60
Soluble-carbohydrate 63.6 76.1
Neutral detergent fiber 458.50 321.20
Acid detergent fiber 266.65 231.05
Lactic acid 3.2 27.4
Acetate 3.535 10.05
Propionic acid 0.162 0.160
Butyric acid 0.085 0.116
Crude protein 201.80 216.20
Ammonia-state nitrogen (NH
3-N) 4.3 2.1
Claims (3)
1. composite green storage inoculating agent; it is characterized in that having adopted that streptococcus uberis and lactobacillus are inoculated simultaneously, asynchronous fermentation technique; tunning is added protective material; with cellulase compounding; be prepared into the powder composite green storage inoculating agent by absorption of normal temperature carrier and post-treatment technology, milk-acid bacteria content is not less than 3 * 10 in the product
10Cfu/g, cellulase activity are not less than 5000 units/g, and the activity of milk-acid bacteria survival rate and cellulase is greater than 80% in 1 year product of stored refrigerated.
2. the preparation method of the described composite green storage inoculating agent of claim 1 is characterized in that:
(1) bacterial classification
With streptococcus uberis as initial fermented bacterium, with lactobacillus as follow-up fermented bacterium;
(2) zymotechnique
The composition of seed culture medium
With the MRS substratum: peptone 10g, meat extract 10g, yeast extract paste 5g, K
2HPO
42g, dibasic ammonium citrate 2g, sodium acetate 5g, glucose 20g, tween 80 1ml, MgSO
47H
2O 0.58g, MnSO
44H
2O 0.25g is settled to 1 with distilled water and is upgraded to the basis, in addition in addition quality percentage meter: tomato juice 5-20%, and Radix Dauci Sativae juice 1-15%, aminothiopropionic acid hydrochloride 0.02-1.0%, accent pH are 6.0-6.8,115 ℃ of steam sterilizings 20 minutes;
The suitability for industrialized production fermention medium is formed
In the quality percentage: rice glucose syrup 3-5% or maize dextrose syrup 3-8%, yeast 0.5-5%, all the other quality percentage components are water; Mineral substance is formed, and the content in every liter of substratum is in g: K
2HPO
41-5g, dibasic ammonium citrate 1-6g, sodium acetate 2-8g, MgSO
47H
2O 0.3-1g, MnSO
44H
2O 0.1-1g;
Culture condition
Seed culture: adopt inoculation, asynchronous fermentation technique simultaneously, streptococcus uberis and lactobacillus are inserted 10 liters of seeding tanks simultaneously cultivate, adopt seed culture medium, each bacterial classification inoculation amount is 4-8%, and 35-40 ℃ of incubator cultivated 18 hours;
Industrial fermentation is produced: seed culture fluid is inoculated in the fermention medium by the 4-8% inoculum size, in 150 liters of fermentor tanks, carry out liquid fermenting, leavening temperature 35-40 ℃, ferment tank pressure 0.02-0.04Mpa, stirring velocity 30-120rpm, initial pH6.0-6.5, fermentation time 16-23 hour;
(3) the composite green storage inoculating agent is composite
Centrifugal bacterium mud and the protective material that obtains of compound lactobacillus fermented liquid that suitability for industrialized production is obtained mixes, bacterium mud and protectant mass volume ratio are 1: 1-4, and protectant quality percentage prescription is: 5-20% skim-milk, 1-8% trehalose, 5-25% glycerine, all the other are water;
Add cellulase then and continue to be mixed into evenly, the mass ratio of bacterium mud and cellulase is 1: 0.5-1;
The absorption carrier gac and the attapulgite that add dry sterilization at last, 1 part of bacterium mud need add 0.3-1.5 part gac and 0.5-1.2 part attapulgite, and fully mixing is crossed the 18-25 mesh sieve and is obtained the good powder of free-running property;
(4) keep-alive post-treatment: the powder that makes is sealed packing with aluminium foil bag,, or vacuumize the room temperature preservation 0-4 ℃ of stored refrigerated.
3. the application of the described composite green storage inoculating agent of claim 1, the addition that it is characterized in that the composite green storage inoculating agent is the 0.5-0.6 ‰ of silage quality, and it is evenly added in the silage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100658084A CN1261561C (en) | 2004-12-15 | 2004-12-15 | Process for preparing high active powder composite green storage inoculating agent and its carrier adsorption agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100658084A CN1261561C (en) | 2004-12-15 | 2004-12-15 | Process for preparing high active powder composite green storage inoculating agent and its carrier adsorption agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1648237A true CN1648237A (en) | 2005-08-03 |
| CN1261561C CN1261561C (en) | 2006-06-28 |
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ID=34868896
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100658084A Expired - Fee Related CN1261561C (en) | 2004-12-15 | 2004-12-15 | Process for preparing high active powder composite green storage inoculating agent and its carrier adsorption agent |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101427739B (en) * | 2008-12-10 | 2011-02-02 | 中国科学院亚热带农业生态研究所 | Green feed storing method |
| CN105063007A (en) * | 2015-07-16 | 2015-11-18 | 江苏天晟药业有限公司 | Method for adsorbing enterococcus by aid of attapulgite |
| CN105331564A (en) * | 2015-12-14 | 2016-02-17 | 甘肃省科学院生物研究所 | Immobilized complex microbial agent and method of directly utilizing immobilized complex microbial agent for fermenting sweet sorghum straw to make feedstuff |
| CN106854626A (en) * | 2015-12-08 | 2017-06-16 | 内蒙古沃德生物质科技有限公司 | A kind of maize straw is blue or green（It is yellow）Store the preparation method of enzyme bacterium composite ferment |
-
2004
- 2004-12-15 CN CNB2004100658084A patent/CN1261561C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101427739B (en) * | 2008-12-10 | 2011-02-02 | 中国科学院亚热带农业生态研究所 | Green feed storing method |
| CN105063007A (en) * | 2015-07-16 | 2015-11-18 | 江苏天晟药业有限公司 | Method for adsorbing enterococcus by aid of attapulgite |
| CN105063007B (en) * | 2015-07-16 | 2018-11-23 | 江苏天晟药业股份有限公司 | A kind of recessed native adsorption method to Lactococcus |
| CN106854626A (en) * | 2015-12-08 | 2017-06-16 | 内蒙古沃德生物质科技有限公司 | A kind of maize straw is blue or green（It is yellow）Store the preparation method of enzyme bacterium composite ferment |
| CN105331564A (en) * | 2015-12-14 | 2016-02-17 | 甘肃省科学院生物研究所 | Immobilized complex microbial agent and method of directly utilizing immobilized complex microbial agent for fermenting sweet sorghum straw to make feedstuff |
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| Publication number | Publication date |
|---|---|
| CN1261561C (en) | 2006-06-28 |
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