CN108018043A - The preparation method and clostridium butyricum composite bacteria agent of a kind of clostridium butyricum composite bacteria agent - Google Patents
The preparation method and clostridium butyricum composite bacteria agent of a kind of clostridium butyricum composite bacteria agent Download PDFInfo
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- CN108018043A CN108018043A CN201711353812.4A CN201711353812A CN108018043A CN 108018043 A CN108018043 A CN 108018043A CN 201711353812 A CN201711353812 A CN 201711353812A CN 108018043 A CN108018043 A CN 108018043A
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- C—CHEMISTRY; METALLURGY
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The present invention provides the preparation method and clostridium butyricum composite bacteria agent of a kind of clostridium butyricum composite bacteria agent, belong to microbial bacteria and technical field, the described method comprises the following steps：1) to soak dregs of beans clostridium butyricum tunning is obtained as matrix anaerobic fermentation clostridium butyricum；2) the clostridium butyricum tunning, NB fluid nutrient mediums are mixed with lactobacillus plantarum and carries out mixed fermentation acquisition clostridium butyricum lactobacillus plantarum tunning；3) the clostridium butyricum lactobacillus plantarum tunning, bacillus and nutrient solution are mixed and obtains clostridium butyricum composite bacteria agent.The clostridium butyricum composite bacteria agent active constituent content that the method prepares is high, can be effectively promoted microorganism growth and decompose organic matter, so as to improve soil environment, plays protection plant and promotes plant growth.
Description
Technical field
The invention belongs to microbial bacterial agent technical field, and in particular to a kind of clostridium butyricum composite bacteria agent preparation method and fourth
Sour clostridium composite bacteria agent.
Background technology
Clostridium butyricum is present in soil, leaf, humans and animals enteron aisle, belongs to beneficial bacterium, has and maintains enteric microorganism
Balance, improve immunity, anticancer, produce the functions such as nutriment.Lactobacillus plantarum belongs to one kind in lactic acid bacteria, plant breast bar
Bacterium can largely produce acid, stablize pH value in water and do not raise, and the acidic materials of its output can degrade heavy metal.Gemma bar
Bacterium, a section for bacterium, can form the bacillus or coccus of gemma (endospore), strong to extraneous injurious factor resistance, and distribution is wide,
It is present in soil, water, air and animal intestinal tract etc..
Humus in soil is that organic matter converts the colloidal substance to be formed through microorganism decomposition, generally black or dark palm fibre
Color, is the chief component of the soil organism.Have in the prior art and micro- life in soil is promoted using clostridium butyricum composite bacteria agent
Thing grows and decomposes organic matter, so as to improve soil environment, plays the role of protecting plant and promotes plant growth.But at present
Market clostridium butyricum compound bacteria product is uneven, and price is high and effect is unstable.
The content of the invention
In view of this, it is an object of the invention to provide a kind of preparation of efficient, low cost clostridium butyricum composite bacteria agent
Method and clostridium butyricum composite bacteria agent, the growth of promotion microorganism and decompose organic matter that the clostridium butyricum can be stablized, so that
Improve soil environment, play protection plant and promote plant growth.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：A kind of system of clostridium butyricum composite bacteria agent
Preparation Method, comprises the following steps：1) clostridium butyricum progress anaerobic fermentation is inoculated with as matrix to soak dregs of beans, obtains clostridium butyricum hair
Ferment product；2) the clostridium butyricum tunning, NB fluid nutrient mediums are mixed with lactobacillus plantarum and carries out mixed fermentation, obtained
Clostridium butyricum-lactobacillus plantarum tunning；3) by the clostridium butyricum-lactobacillus plantarum tunning, bacillus and
Nutrient solution mixes, and obtains clostridium butyricum composite bacteria agent.
Preferably, the water content of the wetting dregs of beans described in step 1) is 8~12wt%.
Preferably, anaerobic fermentation described in step 1) inoculation clostridium butyricum volume for wetting Soybean Meal 12~
16%.
Preferably, the time of anaerobic fermentation clostridium butyricum described in step 1) is 20~28h；The anaerobic fermentation butyric acid shuttle
The temperature of bacterium is 28~32 DEG C.
Preferably, the ratio of lactobacillus plantarum described in step 2) and the viable count of clostridium butyricum in clostridium butyricum tunning
Example is 1:(0.8~1.2).
Preferably, the volume of NB fluid nutrient mediums described in step 2) is 9~12 times of clostridium butyricum tunning.
Preferably, the temperature of mixed fermentation described in step 2) is 33~39 DEG C；The time of the mixed fermentation for 22~
26h。
Preferably, in the clostridium butyricum composite bacteria agent described in step 3) volumn concentration of bacillus for 4.5~
5.5%.
Preferably, the volumn concentration of nutrient solution is 45 in the step 3) the clostridium butyricum composite bacteria agent
~55%.
Present invention also offers the clostridium butyricum composite bacteria agent that above-mentioned preparation method prepares, the clostridium butyricum are compound
The viable count of clostridium butyricum and lactobacillus plantarum is independently (10.5~11) × 10 in microbial inoculum7CFU/g。
Beneficial effects of the present invention：The preparation method of clostridium butyricum composite bacteria agent provided by the invention, operating procedure subtract list,
Raw material dregs of beans is of low cost；Clostridium butyricum tunning is mixed with lactobacillus plantarum after anaerobic fermentation clostridium butyricum
Fermentation, clostridium butyricum and lactobacillus plantarum symbiosis, promote mutually growth, clostridium butyricum active bacteria number increase by 70%, lactobacillus plantarum
Viable count increase by 58%；The clostridium butyricum composite bacteria agent active constituent content that the method prepares is high, each effectively bacterium energy
Enough it is present in as what dominant bacteria was stablized in the soil under natural environment, can stablizes and be effectively promoted microorganism growth and decompose
Organic matter, so as to improve soil environment, plays protection plant and promotes plant growth.
Embodiment
The present invention provides a kind of preparation method of clostridium butyricum composite bacteria agent, comprise the following steps：1) to soak dregs of beans
Clostridium butyricum tunning is obtained for matrix anaerobic fermentation clostridium butyricum；2) the clostridium butyricum tunning, NB liquid are trained
Support base and progress mixed fermentation acquisition clostridium butyricum-lactobacillus plantarum tunning is mixed with lactobacillus plantarum；3) by the fourth
Sour clostridium-lactobacillus plantarum tunning, bacillus and nutrient solution mixing obtain clostridium butyricum composite bacteria agent.
In the present invention, to soak dregs of beans as matrix, inoculation clostridium butyricum carries out anaerobic fermentation, obtains clostridium butyricum fermentation
Product.In the present invention, the water content of the wetting dregs of beans is preferably 8~12wt%, more preferably 9~11wt%, most
Preferably 10wt%.The wetting dregs of beans mixes acquisition with water preferably through by dry dregs of beans in the present invention, described mixed
Conjunction mode preferably sprays application to the water of respective amount in dry dregs of beans.The present invention does not have special limit to the source of the dry dregs of beans
It is fixed, using commercially available dry dregs of beans.
In the present invention, the volume of the clostridium butyricum of anaerobic fermentation inoculation be preferably soak Soybean Meal 12~
16%, more preferably 13~15%, most preferably 14%.The clostridium butyricum of the inoculation preferably uses in the present invention
Bacterium solution after the clostridium butyricum bacterium powder activation of Xiamen Chang Ke bioengineering Co., Ltd production.
The present invention starts the anaerobic fermentation clostridium butyricum after clostridium butyricum to be inoculated in wetting dregs of beans；In the present invention
The mode of the anaerobic fermentation clostridium butyricum preferably stands anaerobic fermentation, and the time of the anaerobic fermentation clostridium butyricum is preferred
For 20~28h, more preferably 22~26h, most preferably 24h；The temperature of heretofore described anaerobic fermentation clostridium butyricum
Preferably 28~32 DEG C, more preferably 30 DEG C of degree.Heretofore described clostridium butyricum can promote microorganism decomposition organic
Thing, forms humus, so as to reach the effect for improving soil, promotes the absorption and growth of Plant To Nutrient material.
The present invention after clostridium butyricum tunning is obtained, by the clostridium butyricum tunning, NB fluid nutrient mediums and
Lactobacillus plantarum mixing carries out mixed fermentation and obtains clostridium butyricum-lactobacillus plantarum tunning.In the present invention, the NB liquid
Body culture medium uses the NB fluid nutrient mediums of this area routine, can specifically use commercially available NB fluid nutrient mediums.At this
The ratio of the viable count of clostridium butyricum is preferably 1 in lactobacillus plantarum described in invention and clostridium butyricum tunning:(0.8
~1.2), it is more preferably 1：1；The volume of the NB fluid nutrient mediums be preferably the clostridium butyricum tunning 9~
12 times, more preferably 10 times.
The mode of the mixed fermentation preferably concussion fermentation in the present invention, the concussion fermentation, which preferably uses, shakes
Bed carries out；The present invention is not particularly limited the shaking table, using the shaking table of this area routine；Heretofore described mixing
The temperature of fermentation is preferably 33~39 DEG C, more preferably 34~38 DEG C, most preferably 35~37 DEG C；The mixed fermentation
The rotating speed of middle shaking table is preferably 150~250rpm, and more preferably 180rpm, the time of the mixed fermentation is preferably 22
~26h, more preferably 24h.Heretofore described lactobacillus plantarum can largely produce acid, pH stable can be maintained not raise,
And the acidic materials of its output can degrade heavy metal；The effect of the mixed fermentation is by the clostridium butyricum and plant breast bar
Bacterium is mixed, the two symbiosis, and promotion is bred each other, improves the viable count of two kinds of bacterium in tunning.
The present invention sends out the clostridium butyricum-lactobacillus plantarum after clostridium butyricum-lactobacillus plantarum tunning is obtained
Ferment product, bacillus and nutrient solution mixing obtain clostridium butyricum composite bacteria agent.In the present invention, it is preferred to first by the butyric acid
Clostridium-lactobacillus plantarum tunning is mixed with bacillus obtain mixed bacteria liquid after, by the mixed bacteria liquid and nutrient solution
Mixing obtains clostridium butyricum composite bacteria agent.The bacillus preferably uses commercially available bacillus in the present invention；It is described
Clostridium butyricum composite bacteria agent in the volumn concentration of bacillus be preferably 4.5~5.5%, more preferably 5%.This
Bacillus described in invention has soil protection, the decomposition for promoting organic matter, produces abundant metabolin and suppresses harmful bacteria
Deng effect, so as to play the role of the growth and protection to plant.
Heretofore described nutrient solution is preferably plant growth nutrient fluid, more preferably commercially available Aron-
Hoagland nutrient solutions；The volumn concentration of nutrient solution is preferably 45 in heretofore described clostridium butyricum composite bacteria agent
~55%, more preferably 50%.
Present invention also offers the clostridium butyricum composite bacteria agent that above-mentioned preparation method prepares, the clostridium butyricum are compound
The viable count of clostridium butyricum and lactobacillus plantarum is independently (10.5~11) × 10 in microbial inoculum7CFU/g.Heretofore described fourth
Sour clostridium composite bacteria agent preparation method is simple, convenient；Cost is low, active constituent content is high；Soil Microorganism can be promoted to grow
With decomposition organic matter, so as to improve soil environment, play the role of protecting plant and promote plant growth.
Clostridium butyricum composite bacteria agent of the present invention can be used as soil conditioner to be applied in soil improvement, in natural ring
In soil under border, it can stablize and be effectively promoted microorganism growth and decompose organic matter, so as to improve soil environment, play guarantor
Protect plant and promote plant growth.
A kind of clostridium butyricum composite bacteria agent preparation method provided by the invention and clostridium butyricum are answered with reference to embodiment
Close microbial inoculum to be described in detail, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
(1) water of 10g dregs of beans 1ml is taken to soak, the clostridium butyricum strain for adding 1.4ml thereto (comes from Xiamen long day
The clostridium butyricum bacterium powder of bioengineering Co., Ltd of section production, the bacterium solution after activation) carry out standing anaerobic fermentation, fermentation temperature 30
DEG C, time 24h.The viable count of clostridium butyricum is 10.8x10 in tunning after fermentation7CFU/g。
(2) whole tunnings after fermenting are added in 100ml NB fluid nutrient mediums (100ml water adds 1.5gNB),
And lactobacillus plantarum is added, additive amount 10.8x107CFU/g, is placed in 35 DEG C of shaking tables mixed fermentation 24h, rotating speed 180rpm；
The viable count of clostridium butyricum and lactobacillus plantarum increases by 70% and 58% respectively after mixed fermentation.
(3) by the bacterium solution after culture, the bacillus of volume 5% is added, composite bacteria liquid is made.
(4) 1 is pressed:1 volume ratio, composite bacteria liquid and Aron-Hoagland nutrient solutions are mixed, compound up to clostridium butyricum
Microbial inoculum.
Embodiment 2
(1) water of 15g dregs of beans 1.2ml is taken to soak, the clostridium butyricum strain for adding 2.1ml thereto (comes from Xiamen
The clostridium butyricum bacterium powder of Chang Ke bioengineering Co., Ltd production, the bacterium solution after activation) carry out standing anaerobic fermentation, fermentation temperature
30 DEG C, time 26h.The viable count of clostridium butyricum is 10.6x10 in tunning after fermentation7CFU/g。
(2) whole tunnings after fermenting are added in 100mlNB fluid nutrient mediums (100ml water adds 1.5gNB),
And lactobacillus plantarum is added, additive amount 10.8x107CFU/g, is placed in 37 DEG C of shaking tables mixed fermentation 22h, rotating speed 180rpm；
The viable count of clostridium butyricum and lactobacillus plantarum increases by 68% and 60% respectively after mixed fermentation.
(3) by the bacterium solution after culture, the bacillus of volume 5% is added, composite bacteria liquid is made.
(4) 1 is pressed:1 volume ratio, composite bacteria liquid and Aron-Hoagland nutrient solutions are mixed, compound up to clostridium butyricum
Microbial inoculum.
Embodiment 3
(1) water of 9g dregs of beans 1ml is taken to soak, the clostridium butyricum strain for adding 1.4ml thereto (comes from Xiamen long day section
The clostridium butyricum bacterium powder of bioengineering Co., Ltd production, the bacterium solution after activation) carry out standing anaerobic fermentation, fermentation temperature 30
DEG C, time 22h.The viable count of clostridium butyricum is 10.8x10 in tunning after fermentation7CFU/g。
(2) whole tunnings after fermenting are added in 100mlNB fluid nutrient mediums (100ml water adds 1.5gNB),
And lactobacillus plantarum is added, additive amount 10.8x107CFU/g, is placed in 36 DEG C of shaking tables mixed fermentation 26h, rotating speed 200rpm；
The viable count of clostridium butyricum and lactobacillus plantarum increases by 71% and 59% respectively after mixed fermentation.
(3) by the bacterium solution after culture, the bacillus of volume 5% is added, composite bacteria liquid is made.
(4) 1 is pressed:1 volume ratio, composite bacteria liquid and Aron-Hoagland nutrient solutions are mixed, compound up to clostridium butyricum
Microbial inoculum.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of preparation method of clostridium butyricum composite bacteria agent, comprises the following steps：
1) clostridium butyricum progress anaerobic fermentation is inoculated with as matrix to soak dregs of beans, obtains clostridium butyricum tunning；
2) the clostridium butyricum tunning, NB fluid nutrient mediums are mixed with lactobacillus plantarum and carries out mixed fermentation, obtain fourth
Sour clostridium-lactobacillus plantarum tunning；
3) clostridium butyricum-lactobacillus plantarum tunning, bacillus and nutrient solution are mixed, obtains clostridium butyricum and answer
Close microbial inoculum.
2. preparation method according to claim 1, it is characterised in that the water content of the wetting dregs of beans described in step 1) is
8~12wt%.
3. preparation method according to claim 1 or 2, it is characterised in that described in step 1) in anaerobic fermentation, the fourth
The inoculation volume of sour clostridium is the 12~16% of wetting Soybean Meal.
4. preparation method according to claim 1, it is characterised in that the time of anaerobic fermentation described in step 1) for 20~
28h；The temperature of the anaerobic fermentation is 28~32 DEG C.
5. preparation method according to claim 1, it is characterised in that lactobacillus plantarum described in step 2) and clostridium butyricum
The ratio of the viable count of clostridium butyricum is 1 in tunning:(0.8~1.2).
6. preparation method according to claim 1 or 5, it is characterised in that the body of NB fluid nutrient mediums described in step 2)
Product is 9~12 times of clostridium butyricum tunning volume.
7. the preparation method stated according to claim 6, it is characterised in that the temperature of mixed fermentation described in step 2) is 33~39
℃；The time of the mixed fermentation is 22~26h.
8. preparation method according to claim 6, it is characterised in that in the clostridium butyricum composite bacteria agent described in step 3)
The volumn concentration of bacillus is 4.5~5.5%.
9. preparation method according to claim 1, it is characterised in that the clostridium butyricum composite bacteria agent described in the step 3)
The volumn concentration of middle nutrient solution is 45~55%.
10. the clostridium butyricum composite bacteria agent that the preparation method described in claim 1~8 any one prepares, its feature exist
In, in the clostridium butyricum composite bacteria agent viable count of clostridium butyricum and lactobacillus plantarum be independently (10.5~11) ×
107CFU/g。
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241053A (en) * | 2019-07-17 | 2019-09-17 | 厦门昶科生物工程有限公司 | A kind of method of mixing fermentation culture clostridium butyricum |
| CN110484468A (en) * | 2019-08-20 | 2019-11-22 | 集美大学 | A compound bacterial agent that improves the adhesion of Clostridium butyricum in the intestinal tract of eel |
| CN110862805A (en) * | 2019-12-03 | 2020-03-06 | 江苏启能新能源材料有限公司 | Constant-temperature phase-change energy storage material and preparation method thereof |
| CN116162461A (en) * | 2023-01-06 | 2023-05-26 | 广西中烟工业有限责任公司 | Alkaline tobacco planting soil bioremediation agent and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110241053A (en) * | 2019-07-17 | 2019-09-17 | 厦门昶科生物工程有限公司 | A kind of method of mixing fermentation culture clostridium butyricum |
| CN110484468A (en) * | 2019-08-20 | 2019-11-22 | 集美大学 | A compound bacterial agent that improves the adhesion of Clostridium butyricum in the intestinal tract of eel |
| CN110862805A (en) * | 2019-12-03 | 2020-03-06 | 江苏启能新能源材料有限公司 | Constant-temperature phase-change energy storage material and preparation method thereof |
| CN116162461A (en) * | 2023-01-06 | 2023-05-26 | 广西中烟工业有限责任公司 | Alkaline tobacco planting soil bioremediation agent and preparation method thereof |
| CN116162461B (en) * | 2023-01-06 | 2025-10-24 | 广西中烟工业有限责任公司 | Alkaline tobacco-growing soil bioremediation agent and preparation method thereof |
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Application publication date: 20180511 |