CN103555593A - Preparation method of Aspergillus niger strain and spore powder - Google Patents
Preparation method of Aspergillus niger strain and spore powder Download PDFInfo
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Abstract
The invention belongs to the field of microbe products, and particularly relates to a preparation method of an Aspergillus niger strain and a spore powder thereof. The strain is Aspergillus niger SDKYSW-2# in Aspergillus, and the collection number is CGMCC No.8327. The preparation method of the Aspergillus niger strain spore powder comprises the following steps: activating the strain, culturing a seed fluid, fermenting and carrying out after-treatment. The method has the advantages of mature production technique, advanced technology, especially advanced strain screening and preparation formation, cheap and accessible raw materials for production, high added value of the product, and obvious ecological and social benefits, and has the obvious characteristics of high benefit, low cost, no environment pollution and no harm to both human and livestock in a microbial fertilizer.
Description
Technical field
The invention belongs to microbial product field, be specifically related to the preparation method of a kind of Aspergillus niger strain and spore powder thereof.
Background technology
Oneself becomes the theme of various countries' agricultural development human survival and Agricul tural Sustain able Development.The predation formula function mode of the high investment high production of modern agriculture (enrich liquid manure, improve the breed, intensive cultivating, chemistryization, mechanize) has illustrated it is extremely harmful.Facts have proved, traditional agriculture and agricultural modernization are combined closely and be just conducive to human survival and development.Since the eighties, American proposed plant growth-promoting rhizobacteria, some rhizospheric microorganism can be affirmed the promoter action of crop, specifically acts on and mainly contains 4 aspects: the perviousness that 1. affects root of the crop cell; 2. affect rhizosphere Metabolic activity; 3. the sorption of microorganism to root secretion; 4. change the validity of nutritive substance to plant.
No matter be the interaction between microorganism, or microorganism is all worked as medium by various enzymes and meta-bolites to the effect of plant.Although various countries scientific worker has done large quantity research to the Ecology of plant growth-promoting rhizobacteria, growth-promoting bacterium by which kind of medium is had an effect to crop and pathogenic bacteria, there is no definite research.Therefore, further investigation plant growth-promoting rhizobacteria meta-bolites has very important meaning to the biological activity of crop and molecular structure feature thereof.Mechanism of action that on the one hand can clear and definite plant growth-promoting rhizobacteria, has very important meaning to bionical synthetic and microbiological genetic engineering transformation on the other hand.Succeeding in developing of positive regulation microbial inoculum particularly, theoretical with in fact all will have important breakthrough.
Summary of the invention
The object of this invention is to provide a kind of Aspergillus niger strain, this bacterial classification can promote growing of crop; The present invention provides the preparation method of Aspergillus niger strain spore powder simultaneously, simple, is applicable to suitability for industrialized production.
Aspergillus niger strain of the present invention is the aspergillus niger SDKYSW-2# in Aspergillus (Aspergillus), Aspergillus niger strain is on October 12nd, 2013, preservation is to China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, Classification And Nomenclature is aspergillus niger (Aspergillusniger), and preserving number is CGMCC No.8327.
Aspergillus niger strain is from booth (Yiyuan) nursery soil, to adopt the separated acquisition of soil isolation by dilution method.
The screening method of Aspergillus niger strain of the present invention, comprises the following steps:
One, soil sample collection: the sampling 200g from upper soll layer to 15cm, the three unities is adopted several samples, mixes scalping, gets 25g and is used as separated;
Two, the separation of aspergillus niger:
1, instrument:
High-pressure steam sterilizing pan, drying baker, incubator, electronic balance, enamelled cup, beaker, packing funnel, test tube, suction pipe (10ml), suction that ball, glass stick, cotton, cotton rope, newspaper, culture dish, triangular flask (100ml), granulated glass sphere, pipettor (1ml), suction nozzle, medicine spoon, inoculating needle, spirit lamp.
2, medicine:
Zulkovsky starch, potassium primary phosphate, sal epsom, ferrous sulfate, agar, sucrose, aspergillus niger.
3, the preparation of aspergillus niger isolation medium:
Zulkovsky starch 10g, potassium primary phosphate 0.5g, sal epsom 0.1g, ferrous sulfate 0.1g, agar 10g, sucrose 10g, water 500ml; Reagent is weighed up, add water and put into enamelled cup, heating for dissolving on electric furnace, inhales in substratum 10ml packing test tube with suction pipe (10ml), has filled in tampon, and paper using is wrapped sterilizing, standby.
Substratum is made: reagent → dissolvings → correction pH value → packing → add tampon, wrapping → sterilizing (0.1P is incubated 30 minutes) → standby.
4, working method:
(1) preparation work:
To use instrument culture dish, triangular flask, suction nozzle (some), medicine spoon, substratum, 10ml test tube etc., with high-pressure sterilizing pot 0.1p pressurize 30 minutes, sterilizing was standby.
Triangular flask: pack 100ml water into and add granulated glass sphere (some).
10ml test tube: pack 9ml water into, carry out numbering, filled in tampon, 8, standby.
5, culture dish is one group, wraps 6 30 of bags with newspaper.
1ml suction nozzle, each wraps (some) with newspaper separately.
(2) dilution of bacterium sample is separated:
Bacterium sample to be separated, under aseptic condition, is got to 1-2 ring with inoculating needle, put into triangular flask, in bottle, bacterium liquid is approximately containing ten thousand/ml of spore 50-100, and shake well, about 5 minutes, mixes each and every one separation of spore, standby.
In inoculation tank by aseptic technique requirement, with pipettor, draw bacterium liquid in 1ml triangular flask and put into pipe No. 1, from No. 1 pipe, inhale 1ml again and put into pipe No. 2, by that analogy, until be diluted to No. 6 or No. 7 in vitro, require every past test tube to inhale and once change suction nozzle one time, in every milliliter of diluent, approximately contain between spore count 5-10.
Get in vitro 0.5 milliliter of bacterium liquid No. 5 No. 6 No. 7, put into culture dish, each extent of dilution is made 10 flat boards, and substratum (temperature 50 C) is poured in culture dish and fully mixed with bacterium liquid, sets level, cooling.
Within about about 20 minutes, after culture medium solidifying in culture dish, take out, be inverted in incubator (30 ℃), cultivate after 4 days, can on plate substratum, observe and take the bacterium colony that each spore grows as concentric(al) circles, the large and spore density of bacterium colony is thick screens good bacterial strain.
Precaution:
This operation must be carried out in sterilized aseptic inoculation box, and strictly by aseptic technique requirement, is undertaken.
The quantity of the culture dish specification of using and the substratum adding must be identical, so that the bacterium colony in different culture dish is compared.
Add the temperature of substratum unsuitable overheated or excessively cold, general control (touches non-scald on hand with hand) otherwise can make spore scald extremely or occur the uneven phenomenon of media surface between 45-50 ℃.
Test tube of every dilution, changes suction nozzle one time.
(3) sieve again triangular flask culture medium: inorganic salt solution is joined in siccative, until the water content of batching is while being 53-55wt.%, stop adding inorganic salt solution; The mass percent of described siccative consists of: wheat bran 50%, soybean cake powder 30% and peanut straw powder 20%; Described inorganic salt solution is containing 2.0gKH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoCl
2.Pack triangular flask into, 500ml triangular flask packs the wet feed 40g of moisture 85% into, and sterilizing accesses the Aspergillus niger strain of above-mentioned separation, cultivates cryodryings after three days for 30 ℃, analyzes the enzyme system producing, in Table 1.
The vigor analysis of the various enzymes of table 1
| ? | Analytical procedure | Calculating vigor |
| Saccharifying enzymic activity | GB | 127u/g |
| Cellulase activity | GB | 7090u/g |
| Xylanase activity | GB | 915u/g |
| Mannosans enzyme activity | GB | 5379u/g |
Comprehensive analysis chosen an Aspergillus niger strain, called after SDKYSW-2#.
Bacterial classification feature: dibbling is to PDA substratum, and black-koji mould filament diameter 15～20pm, is about 1～3mm, wall thickness and smooth.Globe-roof capsule is formed on top, covers one deck metulae and one deck stigma on it comprehensively, and long on stigma have bunchiness memnonious spherical, diameter 2.5～4.0 μ m.Conidial head is spherical, diameter 700～800 μ m, brown-black.Spreading rapidly, just be white, after become aureus until black heavy fleece shape, the colourless or central slightly tawny in the back side.Some bacterial strains also can be converted into androstene by hydroxyl pregnant sterone.Mycelia is chocolate, and top capsule is spherical greatly, and stigma is double-deck, and conidium is spherical, darkly, chocolate, level and smooth or coarse.Patience to ultraviolet ray and ozone is strong.Mycelia is flourishing, and multi-branched has the many cells fungi of multinuclear.Conidiophore by specialization heavy wall and on the hyphal cell (podocyte) that expands, vertically bear; Conidium, head is as " chrysanthemum ".The mycelia of aspergillus niger, spore present shades of colour conventionally, as: black, brown, green, yellow, orange, brown etc., bacterial classification is different, and color is also different.
The preparation method of Aspergillus niger strain spore powder of the present invention, step is as follows:
(1) actication of culture: Aspergillus niger strain is seeded on slant medium and is cultivated,
(2) cultivation of seed liquor: the bacterial classification having activated is joined in seed culture medium and cultivates and obtain seed liquor,
(3) fermentation: the batching after seed liquor and sterilizing stirs, heat-preservation fermentation, spore is sent out in temperature adjustment;
(4) aftertreatment: low-temperature material that step (3) is obtained is dry obtains work in-process, dry, pulverize to standard sieve mesh after adding anticorrosion antiultraviolet material, and spore is collected, and rechecks, packing and get final product.
Slant medium described in step (1) is wheat bran juice slant medium, and culture temperature is 28-30 ℃, and incubation time is 70-74 hour.
The basic recipe of the wheat bran juice substratum described in step (1): Zulkovsky starch 2%, KH
2pO
40.1%, MgSO
40.03%, (NH
4)
2sO
40.05%, agar 2%, sugar 1%, wheat bran juice 500ml.
The bacterial classification that activation described in step (2) is good and the volume ratio of seed culture medium are 1:8-12.
Culture temperature described in step (2) is 33-36 ℃, and incubation time is 45-48 hour.
While cultivating in step (2), centre every 1 hour delivering oxygen once.
The preparation method of the seed culture medium described in step (2) is as follows:
By wheat germ, wheat bran in mass ratio the ratio of 1:9-11 mix, with 45-50 ℃ of warm water, repeatedly rinse, by the scavenging solution obtaining sterilizing 40-50min under 125-130 ℃, 0.15-0.2Mpa, be cooled to 33-36 ℃, make seed culture medium.
The mass ratio of the seed liquor described in step (3) and batching is 1:10-12.
Heat-preservation fermentation temperature described in step (3) is 32-36 ℃, and the heat-preservation fermentation time is 12-15h; It is 32-38 ℃ that spore temperature is sent out in temperature adjustment, and it is 48-50h that the spore time is sent out in temperature adjustment.
Preparation of batch method described in step (3) is as follows:
Inorganic salt solution is joined in siccative, until the water content of batching is while being 53-55wt.%, stop adding inorganic salt solution;
The mass percent of described siccative consists of: wheat bran 40-50%, soybean cake powder 30-40% and peanut straw powder 20-30%;
Described inorganic salt solution is containing 2.0gKH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoCl
2.
In step (3), the fermentation finished product of gained can directly be used.
Anticorrosion antiultraviolet material described in step (4) is zinc oxide, and anticorrosion antiultraviolet material and half-finished mass ratio are 1-1.2:10000.
Cryodrying described in step (4) be spring and autumn winter dryness temperature be 32-34 ℃, be 24-36h time of drying; Summer, drying temperature was 34-38 ℃, and be 36-48h time of drying.
Half-finished moisture≤10% described in step (4).
Standard sieve mesh described in step (4) is 120 sieve meshes.
Reinspection in step (4) is according to Q/SHN--2009 testing product quality.
Primary dcreening operation gating is excessively to taking from different root system of plant soil and rotten plant residue sample, with dilution-plate method, carry out single bacterium separated, by a plurality of Aspergillus niger strains that obtain, select growth and breeding fast, further do product enzyme and sieve again culture experiment, by mensuration, produce the vigor of enzyme, selecting the energetic bacterial strain of cytohydrolist preserves, by technical process, produce aspergillus niger spore powder field test, the same sample plot separated flora of sampling after three months, the dependency experiment of carrying out bacterium turn out to be former experimental bacteria determine grow, and constantly improve and test, aspect bacterial strain, promote plant growth, improve Soil structure, strengthen soil fertility, constant product quality, to people and animals, the target of crop and environmental safety.
In the present invention, bacterial strain screening samples in soil and rotten plant residue, but it is high with pathogenic bacteria, to endanger the residual body screening of serious soil and plant probability, and take containing multiple indication composition substratum is primary dcreening operation substratum, chooses active height and accepts or rejects bacterial strain; Multiple sieve considers that the characteristic of bacterial strain also changes fermentation condition within the specific limits, does not mainly affect its growth velocity, and further improving cell wall enzymes activity is underlying condition, and the bacterial strain screening, through further biological activity determination, is determined the performance of bacterial strain.
The new black-koji mould of a strain that the present invention is selected, is also the bacterial strain that a class is produced prozyme, except producing saccharifying enzyme, also can produce amylase, cellulase etc.Under the effect of enzyme, stodgy macro-molecular protein is degraded to peptone, polypeptide and each seed amino acid, Starch Hydrolysis becomes dextrin, oligosaccharides, monose, and can make in auxiliary material the anti-nutrient substance degradeds such as robust fibre, improve nutritive value, health-care effect and digestibility, can be widely used in brewageing, the industry such as food, fodder additives.
The black-koji mould that the present invention screens (spore content 20,000,000,000/gram, one annual survival rate >=85%), be preserved in national DSMZ, and according to great many of experiments, make and preserve and use all novel form very easily, 20000000000/gram water dispersible granules (pH value: 7.3-7.8, moisture content: 6-8%); Aspergillus niger has the function that secretion produces amylase, saccharifying enzyme, citric acid, gluconic acid, gallic acid etc.; Industrial at bio-feritlizer, aspergillus niger has cracking larger molecular organics and indissoluble inorganics, is convenient to Crop utilization, improves Soil structure, strengthens soil fertility, improves the effect of crop yield.From Yield Increase In Cotton effect, black-koji mould has obvious growth-promoting effect of increasing production to cotton, and after single-dose application, stimulation ratio is 13.50～20.1%.
The present invention compared with prior art, has following beneficial effect:
The present invention has possessed that high efficiency, cost are low, free from environmental pollution, the microbial fertilizer notable feature to person poultry harmless.Mature production technology of the present invention, advanced technology, especially aspect bacterial screening and preparation, have advance, and it is cheap and easy to get to produce required raw material, and added value of product is very high, and ecological and social benefit is remarkable.
Embodiment
Below in conjunction with embodiment, the present invention is described further.
Embodiment 1
(1) actication of culture: be stained with gently a small amount of bacterium colony spore with transfering loop and be linked in wheat bran juice slant medium and cultivate, temperature remains on 30 ℃, cultivates 72 hours;
(2) cultivation of seed liquor: the bacterial classification 1kg having activated is added in 1000kg seed culture medium, 36 ℃ of constant temperature culture 48 hours, middle every 1 hour delivering oxygen once, make seed liquor;
(3) fermentation: the batching 2000kg after seed liquor 200kg and sterilizing stirs inoculation, heat-preservation fermentation, spore is sent out in temperature adjustment, and heat-preservation fermentation temperature is 32 ℃, and the heat-preservation fermentation time is 12h; It is 38 ℃ that spore temperature is sent out in temperature adjustment, and it is 48h that the spore time is sent out in temperature adjustment;
(4) aftertreatment: the material that step (3) is obtained, cryodrying obtains work in-process, and 1kg work in-process dry, pulverize to 120 sieve meshes after adding zinc oxide 0.1g, and spore is collected, and rechecks, packing and get final product.
Described in step (3), preparation of batch method is as follows:
Inorganic salt solution is joined in siccative, until the water content of batching is while being 53wt.%, stop adding inorganic salt solution;
Described siccative is wheat bran 1000kg, soybean cake powder 600kg and peanut straw powder 400kg;
Described inorganic salt solution is containing 2.0g KH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoC
l2.
Various enzyme activity analysis in table 2 in finished product.
Various enzyme activity analyses in table 2 finished product
| ? | Analytical procedure | Calculating vigor |
| Saccharifying enzymic activity | GB | 98u/g |
| Cellulase activity | GB | 6872u/g |
| Xylanase activity | GB | 891u/g |
| Mannosans enzyme activity | GB | 3582u/g |
Total spore count: 35,000,000,000
Living bacteria count: 14,500,000,000
Embodiment 2
(1) actication of culture: be stained with gently a small amount of bacterium colony spore with transfering loop and be linked in wheat bran juice slant medium and cultivate, temperature remains on 28 ℃, cultivates 70 hours;
(2) cultivation of seed liquor: the bacterial classification 1kg having activated is added in 800kg seed culture medium, 33 ℃ of constant temperature culture 45 hours, middle every 1 hour delivering oxygen once, make seed liquor;
(3) fermentation: the batching 2400kg after seed liquor 200kg and sterilizing stirs inoculation, heat-preservation fermentation, spore is sent out in temperature adjustment, and heat-preservation fermentation temperature is 36 ℃, and the heat-preservation fermentation time is 15h; It is 32 ℃ that spore temperature is sent out in temperature adjustment, and it is 50h that the spore time is sent out in temperature adjustment;
(4) aftertreatment: the material that step (3) is obtained, cryodrying obtains work in-process, and 1kg work in-process dry, pulverize to 120 sieve meshes after adding zinc oxide 0.12g, and spore is collected, and rechecks, packing and get final product.
Described in step (3), preparation of batch method is as follows:
Inorganic salt solution is joined in siccative, until the water content of batching is while being 55wt.%, stop adding inorganic salt solution;
Described siccative is wheat bran 960kg, soybean cake powder 960kg and peanut straw powder 480kg;
Described inorganic salt solution is containing 2.0g KH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoCl
2.
Various enzyme activity analysis in table 3 in finished product.
Various enzyme activity analyses in table 3 finished product
| ? | Analytical procedure | Calculating vigor |
| Saccharifying enzymic activity | GB | 90u/g |
| Cellulase activity | GB | 5882u/g |
| Xylanase activity | GB | 752u/g |
| Mannosans enzyme activity | GB | 3106u/g |
Total spore count: 32,000,000,000
Living bacteria count: 12,500,000,000
Embodiment 3
(1) actication of culture: be stained with gently a small amount of bacterium colony spore with transfering loop and be linked in wheat bran juice slant medium and cultivate, temperature remains on 29 ℃, cultivates 74 hours;
(2) cultivation of seed liquor: the bacterial classification 1kg having activated is added in 1200kg seed culture medium, 35 ℃ of constant temperature culture 46 hours, middle every 1 hour delivering oxygen once, make seed liquor;
(3) fermentation: the batching 2200kg after seed liquor 200kg and sterilizing stirs inoculation, heat-preservation fermentation, spore is sent out in temperature adjustment, and heat-preservation fermentation temperature is 35 ℃, and the heat-preservation fermentation time is 14h; It is 35 ℃ that spore temperature is sent out in temperature adjustment, and it is 49h that the spore time is sent out in temperature adjustment;
(4) aftertreatment: the material that step (3) is obtained, cryodrying obtains work in-process, and 1kg work in-process dry, pulverize to 120 sieve meshes after adding zinc oxide 0.11g, and spore is collected, and rechecks, packing and get final product.
Described in step (3), preparation of batch method is as follows:
Inorganic salt solution is joined in siccative, until the water content of batching is while being 54wt.%, stop adding inorganic salt solution;
Described siccative is wheat bran 880kg, soybean cake powder 660kg and peanut straw powder 660kg.
Described inorganic salt solution is containing 2.0g KH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoCl
2.
Various enzyme activity analysis in table 4 in finished product.
Various enzyme activity analyses in table 4 finished product
| ? | Analytical procedure | Calculating vigor |
| Saccharifying enzymic activity | GB | 92u/g |
| Cellulase activity | GB | 6159u/g |
| Xylanase activity | GB | 786u/g |
| Mannosans enzyme activity | GB | 3386u/g |
Total spore count: 32,500,000,000
Living bacteria count: 13,000,000,000
Claims (10)
1. an Aspergillus niger strain, is characterized in that: this bacterial strain is the aspergillus niger SDKYSW-2# in Aspergillus (Aspergillus), and its preserving number is CGMCC No.8327.
2. a preparation method for Aspergillus niger strain spore powder claimed in claim 1, is characterized in that step is as follows:
(1) actication of culture: Aspergillus niger strain is seeded on slant medium and is cultivated,
(2) cultivation of seed liquor: the bacterial classification having activated is joined in seed culture medium and cultivates and obtain seed liquor,
(3) fermentation: the batching after seed liquor and sterilizing stirs, heat-preservation fermentation, spore is sent out in temperature adjustment;
(4) aftertreatment: low-temperature material that step (3) is obtained is dry obtains work in-process, dry, pulverize to standard sieve mesh after adding anticorrosion antiultraviolet material, and spore is collected, and rechecks, packing and get final product.
3. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the slant medium described in step (1) is wheat bran juice slant medium, and culture temperature is 28-30 ℃, and incubation time is 70-74 hour.
4. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that bacterial classification that the activation described in step (2) is good and the volume ratio of seed culture medium are 1:8-12.
5. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the culture temperature described in step (2) is 33-36 ℃, and incubation time is 45-48 hour.
6. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the preparation method of the seed culture medium described in step (2) is as follows:
By wheat germ, wheat bran in mass ratio the ratio of 1:9-11 mix, with 45-50 ℃ of warm water, repeatedly rinse, by the scavenging solution obtaining sterilizing 40-50min under 125-130 ℃, 0.15-0.2Mpa, be cooled to 33-36 ℃, make seed culture medium.
7. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the mass ratio of the seed liquor described in step (3) and batching is 1:10-12.
8. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the heat-preservation fermentation temperature described in step (3) is 32-36 ℃, and the heat-preservation fermentation time is 12-15h; It is 32-38 ℃ that spore temperature is sent out in temperature adjustment, and it is 48-50h that the spore time is sent out in temperature adjustment.
9. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the preparation of batch method described in step (3) is as follows:
Inorganic salt solution is joined in siccative, until the water content of batching is while being 53-55wt.%, stop adding inorganic salt solution;
The mass percent of described siccative consists of: wheat bran 40-50%, soybean cake powder 30-40% and peanut straw powder 20-30%;
Described inorganic salt solution is containing 2.0gKH in every premium on currency
2pO
4, 1.4g (NH
4)
2sO
4, 0.3g urea, 0.3gMgSO
47H
2o, 0.3g CaCl
2, 5.0mg FeSO
47H
2o, 1.56mg MnSO
4h
2o, 1.4mg ZnSO
47H
2o, 2.0mgCoCl
2.
10. the preparation method of Aspergillus niger strain spore powder according to claim 2, is characterized in that the anticorrosion antiultraviolet material described in step (4) is zinc oxide, and anticorrosion antiultraviolet material and half-finished mass ratio are 1-1.2:10000.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107011029A (en) * | 2017-04-06 | 2017-08-04 | 哈尔滨尼亚农业有限公司 | Seedling agent and its production method are strengthened in a kind of green acid adjustment of high-effective microorganism |
| CN109423450A (en) * | 2017-08-31 | 2019-03-05 | 勐海茶业有限责任公司 | A kind of aspergillus niger microbial inoculum of fermentation of pu'er tea and preparation method thereof |
| CN113430124A (en) * | 2021-06-29 | 2021-09-24 | 佛山市海天(高明)调味食品有限公司 | Culture medium for increasing number of soy sauce aspergillus spores and preparation method thereof |
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| CN101591619A (en) * | 2009-07-03 | 2009-12-02 | 中国农业科学院饲料研究所 | Aspergillus niger strain and application thereof |
| CN102987098A (en) * | 2012-12-07 | 2013-03-27 | 青岛根源生物技术集团有限公司 | Special complex enzyme preparation for wheat for livestock and poultry and application thereof |
| CN103374528A (en) * | 2013-08-09 | 2013-10-30 | 牛赡光 | Aspergillus niger strain and application thereof |
-
2013
- 2013-11-07 CN CN201310548819.7A patent/CN103555593B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101591619A (en) * | 2009-07-03 | 2009-12-02 | 中国农业科学院饲料研究所 | Aspergillus niger strain and application thereof |
| CN102987098A (en) * | 2012-12-07 | 2013-03-27 | 青岛根源生物技术集团有限公司 | Special complex enzyme preparation for wheat for livestock and poultry and application thereof |
| CN103374528A (en) * | 2013-08-09 | 2013-10-30 | 牛赡光 | Aspergillus niger strain and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107011029A (en) * | 2017-04-06 | 2017-08-04 | 哈尔滨尼亚农业有限公司 | Seedling agent and its production method are strengthened in a kind of green acid adjustment of high-effective microorganism |
| CN109423450A (en) * | 2017-08-31 | 2019-03-05 | 勐海茶业有限责任公司 | A kind of aspergillus niger microbial inoculum of fermentation of pu'er tea and preparation method thereof |
| CN113430124A (en) * | 2021-06-29 | 2021-09-24 | 佛山市海天(高明)调味食品有限公司 | Culture medium for increasing number of soy sauce aspergillus spores and preparation method thereof |
| CN113430124B (en) * | 2021-06-29 | 2023-02-21 | 佛山市海天(高明)调味食品有限公司 | Culture medium for increasing number of soy sauce aspergillus spores and preparation method thereof |
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| Publication number | Publication date |
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| CN103555593B (en) | 2015-06-10 |
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