CN102816709A - Method for preparing composite biological agent by double-bacterium co-culture - Google Patents
Method for preparing composite biological agent by double-bacterium co-culture Download PDFInfo
- Publication number
- CN102816709A CN102816709A CN2011101501418A CN201110150141A CN102816709A CN 102816709 A CN102816709 A CN 102816709A CN 2011101501418 A CN2011101501418 A CN 2011101501418A CN 201110150141 A CN201110150141 A CN 201110150141A CN 102816709 A CN102816709 A CN 102816709A
- Authority
- CN
- China
- Prior art keywords
- microbes
- cultivated
- bacterium
- bacterial classification
- altogether
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention discloses a method for preparing a composite biological agent by double-bacterium co-culture. The method is characterized in that two microbes with different functionalities are co-cultured in a same fermentation apparatus. According to the invention, with life activities between the two microbes, and produced substances or a formed environment, the microbes are interdependent and are mutually benefited. At a same time, a preparation containing products of two microbes is obtained. According to physiological characteristics of strains, two microbes with different growth speeds and different growth periods are adopted, wherein the microbes can perform nutrition complementation, and can live in a same environment. With a co-culturing process, a product containing two functional microbes and metabolite thereof is obtained. Compared with independent culturing, the method provided by the invention is advantaged in improved production efficiency, reduced production cost, and substantially improved application effect. With the product, soil water-soluble phosphorus is increased, and soil nitrogen fixation capacity is improved.
Description
Technical field
The present invention is specifically related to a kind of pair of bacterium and cultivates the method for preparing composite bacteria agent altogether, belongs to the biological fermentation field, is to adopt two kinds of different functionalities mikrobes; In same fermentation container; Co-cultivation finally obtains a kind of preparation, contains the product of two kinds of mikrobes.
Background technology
Complex microorganism preparations, the product that is considered to have the mikrobe complex function.Two bacterium are cultivated altogether, and its meaning also is both vital movements with each other, the material of generation or the environment that causes, and interdependence benefits each other.This series products is that representative is widely used in fields such as medicine, health care, agricultural, livestock industry with little ecological compound formulation.Combination of two bacterium and common culture condition are the key problem in technology of microbial inoculum quality all the time.Know also there is not the report of relative technological products at present with regard to the contriver.
Summary of the invention
The object of the invention provides a kind of fields such as medicine, health care, agricultural, livestock industry that are widely used in exactly for remedying above deficiency, and its physiologically active has a kind of pair of bacterium of obvious enhanced advantage and cultivates the method for preparing composite bacteria agent altogether.
Main technical schemes is achieved in that two kinds of different functionalities mikrobes of employing, in same fermentation container, and co-cultivation.Rely on two kinds of mikrobes vital movement each other, the material of generation or the environment that causes, interdependence benefits each other.Obtain a kind of preparation simultaneously, contain the product of two kinds of mikrobes.Two bacterium are cultivated the method for preparing composite bacteria agent altogether; Comprise function culture of strains, selection and combination; The control of culture medium prescription and preparation, culture condition and design; It is characterized in that taking the speed of growth respectively and the cycle is different, nutrition complement, two kinds of mikrobes can residing in same environment altogether each other,, obtain to contain the product of two kinds of function yeast and metabolite thereof through cultivating altogether according to the physilogical characteristics of bacterial classification.
Described bacterial classification comprises multiple mikrobes such as bacillus, genus lactubacillus, Saccharomycodes, acetic acid Pseudomonas, Azotobacter, rhizobium.Has the function of producing various active enzyme, multiple organic acid, VITAMINs and fixed nitrogen, phosphorus decomposing, potassium decomposing, pest-resistant, disease-resistant, stimulating growth.Be applied in agricultural, can improve crop alimentary effectively, improve resistance, improve quality, increase output.
Describedly cultivating altogether, is to adopt the general deep liquid culture tank of fermentation industry, selects the substratum of suitable growth according to the needs of each bacterial classification, and aeration condition is cultivated, and thalline reaches maximum quantity, and maturation is put jar.
Described cultivation altogether also can adopt deep liquid to leave standstill cultivation as required, requires to select appropriate media according to each bacterial classification, and mainly be under anaerobic to make its fermentation, the viable bacteria and the tunning of acquisition some amount.
The slant medium that said process for preparing strain thereof is used has following component and preferable weight proportion: peptone 8～12g Carnis Bovis seu Bubali cream 4～6g glucose 1～3g agar 12～20g zero(ppm) water 1L.
The substratum that described bacterial classification liquid culture is used has following component and preferable weight proportion:
Soybean hydrolyzate 10～30g peptone 4～10g sucrose 16～24g KH
2PO
43～8gMgSO
41～6g water 1L.
Described bacterial classification deep liquid leaves standstill the substratum of cultivating usefulness, has following component and preferable weight proportion:
Sucrose 10～30g peptone 4～10g yeast extract paste 5-10g KH
2PO
45～10g MgSO
41～6g water 1L.
Positively effect of the present invention is: in same incubator, cultivate altogether through two bacterium, obtain a kind of product with two kinds of functional microorganisms.Improve production efficiency, practiced thrift production cost greatly.
Compared with prior art: the present invention at first with the mikrobe of two kinds of specific functions, is incubated at same incubator altogether, obtains active somatic cell and the meta-bolites of two kinds of mikrobes simultaneously.With cultivation respectively separately, its physiologically active has obvious enhanced advantage.
Implementation result of the present invention:
Composite bacteria agent and of the influence (inoculation back 10 round the clock) of independent bacillus megaterium to water-soluble phosphorus content in the soil
Experimental result shows, the composite bacteria group is inoculated in behind the soil 10 round the clock, and the soil water-soluble phosphorus is not only considerably beyond contrast, and increased by 30.8% than the soil water-soluble phosphorus of inoculation bacillus megaterium separately.
Composite bacteria agent and of the comparison of independent vinelandii to the influence of soil nitrogen fixing capacity
Composite fungus agent and vinelandii are inoculated in respectively in the soil of sterilized, are contrast not connect bacterium.Warp 5,10, I5, cultivation are round the clock got each processing and control soil 1g and are inoculated into by on the Dobere i ner nitrogen-free agar wetted silica sand (every ware is put silica sand 25g), measure institute's fixed nitrogen amount round the clock the 10th.Experimental result shows, the soil of composite fungus agent inoculation, through 5,10,15, round the clock cultivate institute's fixed nitrogen amount compare respectively according to increase by 75.6%, 89.2% and, 84.2%, increase by 20%, 25% and 22.1% during respectively than independent inoculation vinelandii
Embodiment
Instance 1.
Slant strains: bacillus megaterium 1., vinelandii are the test tube slant 2., in 25-32 ℃ of constant temperature culture, 24-72h.
Bacteria suspension: with 1. 2. test tube slant of bacterial classification, pour sterilized water respectively into, scrape and get lawn, the preparation bacteria suspension.
Bacterial classification mixes: with 1. 2. bacteria suspension of bacterial classification, proportional mixing.
Liquid nutrient medium: the substratum that the bacterial classification liquid culture is used has following component and weight proportion:
Soybean hydrolyzate 10～30g peptone 4～10g sucrose 16～24gKH
2PO
43～8g MgSO
41～6g water 1L.
Sterilization: 0.12-0.15Mpa, 121-125 ℃, steam sterilizing 30 minutes.
Inoculation: bacteria suspension is inserted in the substratum of sterilization.
Cultivate: 25-32 ℃, tank pressure 0.04-0.05Mpa cultivates 36-72h.
Instance 2.
Slant strains: milk-acid bacteria 3., sporeformer 4. the test tube slant in 25-38 ℃ of constant temperature culture, 24-72h.
Bacteria suspension: with 3. 4. test tube slant of bacterial classification, pour sterilized water respectively into, scrape and get lawn, the preparation bacteria suspension.
Bacterial classification mixes: with 3. 4. bacteria suspension of bacterial classification, proportional mixing.
Liquid nutrient medium: sucrose 10～30g peptone 4～10g yeast extract paste 5-10g KH
2PO
45～10g MgSO
41～6g water 1L
Sterilization: 0.12-0.15Mpa, 121-125 ℃, steam sterilizing 30 minutes.
Inoculation: bacteria suspension is inserted in the substratum of sterilization.
Cultivate: 25-32 ℃, cultivate 2-7d.
Claims (5)
1. two bacterium are cultivated the method for preparing composite bacteria agent altogether; Comprise selection, combination and the cultivation of function bacterial classification; The control of culture medium prescription and preparation, culture condition and design; It is characterized in that taking the speed of growth respectively and the cycle is different, nutrition complement, two kinds of mikrobes can residing in same environment altogether each other,, obtain to contain the product of two kinds of function yeast and metabolite thereof through cultivating altogether according to the physilogical characteristics of bacterial classification.
2. pair bacterium according to claim 1 is cultivated the method for preparing composite bacteria agent altogether; Its characteristic adopts to comprise multiple mikrobes such as bacillus, genus lactubacillus, Saccharomycodes, acetic acid Pseudomonas, Azotobacter, rhizobium in the selection that is the function bacterial classification.Has the function of producing various active enzyme, multiple organic acid, VITAMINs and fixed nitrogen, phosphorus decomposing, potassium decomposing, pest-resistant, disease-resistant, stimulating growth.Be applied in agricultural, can improve crop alimentary effectively, improve resistance, improve quality, increase output.Select aerobic training method, two bacterium are cultivated.The inoculative proportion that two bacterium are cultivated: 0.1-2: 1-10.
3. pair bacterium according to claim 2 is cultivated the method for preparing composite bacteria agent altogether, and its characteristic has the ratio of soybean hydrolyzate and each component in the substratum that is the function bacterial classification: soybean hydrolyzate 10～50g peptone 4～1Og sucrose 16～24g KH
2PO
43～8g MgSO
41～6g water 1L.
4. pair bacterium according to claim 2 is cultivated the method for preparing composite bacteria agent altogether; Its characteristic is being that spawn culture adopts bacterial classification in vital movement; Can produce various active enzyme and secondary metabolite, like the mikrobe of multiple organic acid, small-molecular peptides, VITAMINs, oligosaccharide substance.The anaerobism training method of selecting deep layer to leave standstill, compound cultivation.Inoculum size proportioning: 1: 5-40.
5. pair bacterium according to claim 4 is cultivated the method for preparing composite bacteria agent altogether, and its characteristic is being that the anaerobic culture medium electricity that deep layer leaves standstill has following component and weight proportion: sucrose 10～30g peptone 4～10g yeast extract paste 5-10g KH
2PO
45～10g MgSO
41～6g water 1L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101501418A CN102816709A (en) | 2011-06-07 | 2011-06-07 | Method for preparing composite biological agent by double-bacterium co-culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101501418A CN102816709A (en) | 2011-06-07 | 2011-06-07 | Method for preparing composite biological agent by double-bacterium co-culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102816709A true CN102816709A (en) | 2012-12-12 |
Family
ID=47301163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101501418A Pending CN102816709A (en) | 2011-06-07 | 2011-06-07 | Method for preparing composite biological agent by double-bacterium co-culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102816709A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106957870A (en) * | 2017-03-17 | 2017-07-18 | 山东普方生物科技有限公司 | A kind of composite biological agent and application thereof |
| CN108531435A (en) * | 2018-04-10 | 2018-09-14 | 湖南农业大学 | A kind of strain encryption microbial inoculum production method of bacillus subtilis complementation culture |
| CN108690813A (en) * | 2018-05-24 | 2018-10-23 | 鲁东大学 | A kind of preparation method and applications of silicate-dissolving microbe microbial inoculum |
-
2011
- 2011-06-07 CN CN2011101501418A patent/CN102816709A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 龙苏 等: "固氮球形芽孢杆菌与巨大芽孢杆菌的混合增效作用", 《核农学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106957870A (en) * | 2017-03-17 | 2017-07-18 | 山东普方生物科技有限公司 | A kind of composite biological agent and application thereof |
| CN109456900A (en) * | 2017-03-17 | 2019-03-12 | 山东普方生物科技有限公司 | A kind of composite biological agent and application thereof |
| CN109456900B (en) * | 2017-03-17 | 2022-09-20 | 山东普方生物科技有限公司 | Composite biological preparation and application thereof |
| CN108531435A (en) * | 2018-04-10 | 2018-09-14 | 湖南农业大学 | A kind of strain encryption microbial inoculum production method of bacillus subtilis complementation culture |
| CN108531435B (en) * | 2018-04-10 | 2020-09-22 | 湖南农业大学 | Production method of strain encryption microbial inoculum for complementary culture of bacillus subtilis |
| CN108690813A (en) * | 2018-05-24 | 2018-10-23 | 鲁东大学 | A kind of preparation method and applications of silicate-dissolving microbe microbial inoculum |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104974966B (en) | A kind of bacillus coagulans and its fermentation process in high density and dry bacterium powder preparation method | |
| CN102409007B (en) | Bacillus microecological preparation and liquid-solid fermentation combining preparation process thereof | |
| CN105567624B (en) | A kind of the Lactococcus lactis cream subspecies synergist and its application method of the lactic acid producing streptostacin that ferments | |
| CN106011041B (en) | For promoting the compound lactobacillus microbial inoculum and preparation method of Tomato Quality | |
| CN102199050A (en) | Composite microbial fertilizer and preparation method thereof | |
| CN102925504B (en) | Method and fermentation culture medium used for synthesizing gamma-aminobutyric acid through microbial fermentation | |
| Pyar et al. | Potentials of pineapple waste as growth medium for Lactobacillus species | |
| CN107828698A (en) | A kind of composite bacteria preparation and its preparation method and application | |
| CN106929450A (en) | It is a kind of to promote composite bacteria agent of the quick decomposition of agricultural wastes and preparation method thereof | |
| CN106754524A (en) | Lactobacillus paracasei N1115 culture mediums and its application | |
| CN104651267B (en) | A kind of organic fertilizer with the microbial bacteria and its application of fermentation production alkali | |
| CN102994430B (en) | Bacterial cellulose production strain and application thereof | |
| CN103243039A (en) | High density culturing method of lactobacillus paracasei | |
| CN102815973A (en) | Preparation method of biological selenium-rich fertilizer | |
| CN103992977A (en) | Method for culturing bacillus subtilis through high-density fermentation | |
| CN110241053A (en) | A kind of method of mixing fermentation culture clostridium butyricum | |
| CN106497842A (en) | The preparation method and application of a kind of lactic acid bacteria and aspergillus niger mixed solid fermentation preparation | |
| CN102816709A (en) | Method for preparing composite biological agent by double-bacterium co-culture | |
| CN102417890A (en) | Sinorhizobium meliloti and method for applying same for fermenting to produce manganese peroxidase | |
| CN102127515A (en) | Screening and application of L-proline high-producing Brevundimonas sp. (JNPP-1) | |
| CN103289912A (en) | Solid fermentation method of bacillus coagulans | |
| CN103103062A (en) | Method for preparing nutritional mulberry beverage from composite probiotics | |
| CN103849575B (en) | A kind of production method of single cell protein | |
| CN108641977A (en) | A method of mixing high density fermentation improves bifidobacterium cells density and metabolite | |
| CN105018409A (en) | Methyl nutritional type bacillus spore-production fermentation medium and culture method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20121212 |