CN108148783A - A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders - Google Patents
A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders Download PDFInfo
- Publication number
- CN108148783A CN108148783A CN201810095745.9A CN201810095745A CN108148783A CN 108148783 A CN108148783 A CN 108148783A CN 201810095745 A CN201810095745 A CN 201810095745A CN 108148783 A CN108148783 A CN 108148783A
- Authority
- CN
- China
- Prior art keywords
- gemma
- bacillus amyloliquefaciens
- preparation
- culture medium
- production
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
Abstract
The present invention is suitable for technical field of microbial fermentation, provides a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders, and one, prepare production gemma culture medium, production gemma nutrient media components are：Cornstarch, glucose, dregs of beans, calcium carbonate, ammonium chloride, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate；2nd, gemma zymotic fluid is cultivated；3rd, gemma bacterium powder is prepared, ammonium sulfate and diatomite are added into the gemma zymotic fluid；Then plate-frame filtering is carried out；Obtain the gemma bacterium powder of bacillus amyloliquefaciens G5.Whereby, in the preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders provided by the invention, production gemma culture medium raw material is common, is conveniently easy to get, reduces production cost；Number of spores is more in obtained gemma zymotic fluid, gemma rate >=98%；Using ammonium sulfate and diatomite as flocculant, shorten the time prepared by gemma bacterium powder, living bacteria count is more in obtained gemma bacterium powder, yield >=95% of effective viable bacteria.
Description
Technical field
The present invention relates to technical field of microbial fermentation, and in particular to a kind of system of bacillus amyloliquefaciens G5 gemma bacterium powders
Preparation Method.
Background technology
Bacillus amyloliquefaciens are extensive in distributed in nature as a kind of probiotics, nontoxic to people and animals, not dirty
Environment is contaminated, growth is fast, and stability is good, has broad spectrum antibiotic activity and stronger anti-adversity ability.Gemma is also known as endospore, is thin
Bacterium hypopus；Spore-producing bacterium forms round or ellipse gemma hypopus in the cell when nutritional condition lacks.Gemma
Water content is extremely low, and resistance is strong, is amenable to high temperature, ultraviolet light, and ionising radiation and a variety of chemical substances are killed, and condition is suitable
When, gemma can become a bacterium again；It is the bacterium powder system for preparing since gemma is high compared with its easy preservation of nutrition body cell, resurrection rate
The preferable existence form of agent.
At this stage, the preparation method of bacillus amyloliquefaciens gemma bacterium powder is usually that zymotic fluid is directly spray-dried,
Or zymotic fluid is flocculated with disodium hydrogen phosphate, then carries out fluidized drying.Solution starch gemma is prepared with existing preparation method
Bacillus bacterium powder, obtained Number of spores is few, and gemma rate is 90% or so；Living bacteria count is few in bacterium powder, and effective viable bacteria yield is only
It can be 50% or so；Production cost is very high simultaneously.Therefore, the preparation method of bacillus amyloliquefaciens gemma bacterium powder is not at this stage
It is preferable.
In summary, the prior art there will naturally be shortcoming in practical applications, it is necessary to be improved.
Invention content
For above-mentioned technological deficiency, the purpose of the present invention is to provide a kind of systems of bacillus amyloliquefaciens G5 gemma bacterium powders
Preparation Method, this method provide a kind of novel production gemma culture medium, while the culture solution starch gemma on novel production gemma culture medium
The gemma zymotic fluid of bacillus G5, then using ammonium sulfate and diatomite as flocculant, plate-frame filtering is then carried out, obtained solution starch
Bacillus G5 Number of spores is more, and gemma rate is high；Living bacteria count is more in gemma bacterium powder, living bacteria count high income.
To achieve these goals, the present invention provides a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders, packet
Include following steps：
The first step prepares production gemma culture medium
Producing gemma culture medium raw material is：Carbon source, nitrogen source and inorganic salts；
The production gemma culture medium each group is divided into：10~40g/L of cornstarch, 0.5~3g/L of glucose, dregs of beans 5~
25g/L, 0.1~1.5g/L of calcium carbonate, 0.1~1.0g/L of ammonium chloride, 0.1~1.0g/L of dipotassium hydrogen phosphate, magnesium sulfate 0.01~
0.01~0.7g/L of 0.7g/L and manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank, water is added to adjust concentration；It is 7.5~7.8 to adjust pH again,
Then the high-temperature sterilization under the conditions of 121~123 DEG C；Obtain production gemma culture medium；
Second step culture gemma zymotic fluid
Under aseptic condition, the seed liquor of bacillus amyloliquefaciens G5 is transferred in the fermentation tank with volume ratio 5~6%,
The fermented and cultured in the production gemma culture medium；After fermentation, the fermentation tank is warming up to 50~60 DEG C, maintain 20~
30min obtains the gemma zymotic fluid of the bacillus amyloliquefaciens G5；
Third step prepares gemma bacterium powder
Ammonium sulfate and diatomite are added into the gemma zymotic fluid, is stirred evenly；Then plate-frame filtering is carried out, collects filter
Cake；The filter cake is dried with boiling drying equipment again, 25~40min is dried under the conditions of 70~80 DEG C；Finally crushed with pulverizer
The filter cake after drying crosses 40~60 mesh sieve；Obtain the gemma bacterium powder of bacillus amyloliquefaciens G5.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the bacillus amyloliquefaciens G5
For the dominant strain isolated and purified in spoil fermented sample.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the bacillus amyloliquefaciens G5
Seed liquor preparation process include：
Step 1 actication of culture
Under aseptic condition, the bacillus amyloliquefaciens G5 is seeded to activation culture in beef-protein medium,
22~28h is cultivated under the conditions of 36~38 DEG C；It crosses and is forwarded on beef extract-peptone tablet again, separation single bacterium colony, 36~38 DEG C
Under the conditions of cultivate 22~28h, repeatedly activate 2~3 times；
Step 2 prepares seed liquor
Under aseptic condition, single bacterium colony described in picking is forwarded in the beef extract-peptone fluid nutrient medium of sterilizing, 36~38
8~12h is cultivated under the conditions of DEG C, obtains the seed liquor.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the carbon of the production gemma culture medium
Source raw material further includes：Maize flour and sucrose；Nitrogen source raw material further includes：Fish meal and peptone；Inorganic salt raw material further includes：Sulfuric acid
Ammonium, potassium dihydrogen phosphate and ferric trichloride.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the condition of culture of the fermentation tank
For：Under the conditions of 36~38 DEG C cultivate 23~36h, rotating speed be 180~200rpm, ventilation quantity 1：1.2~1.5, tank pressure is 0.05
~0.08Mpa.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the charge of the fermentation tank is
60~70%.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the ammonium sulfate and the diatom
The proportion that soil accounts for the gemma zymotic fluid is respectively：Ammonium sulfate is 30%~40%, and diatomite is 3%~8%.
According to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention, the bud in the gemma zymotic fluid
Spore rate >=98%.
It is effective in the gemma bacterium powder according to a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders of the present invention
Viable bacteria yield >=95%.
The purpose of the present invention is to provide a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders, by providing one
Novel production gemma culture medium is planted, while the gemma zymotic fluid of bacillus amyloliquefaciens G5 is cultivated on novel production gemma culture medium,
Again using ammonium sulfate and diatomite as flocculant, plate-frame filtering is then carried out, obtains bacillus amyloliquefaciens G5 gemma bacterium powders.This
Invention production gemma culture medium raw material is relatively common, is conveniently easy to get, greatly reduces production cost；In fermentation process, zymotechnique
Simply, it is easy to control, it does not need to carry out feed supplement；Number of spores is more in obtained bacillus amyloliquefaciens G5 gemma zymotic fluids, gemma
Rate >=98%；Meanwhile using ammonium sulfate and diatomite as flocculant, shorten the time prepared by bacterium powder, obtained solution starch gemma
Living bacteria count is 4~6 times higher than the prior art in bacillus G5 gemma bacterium powders, yield >=95% of effective viable bacteria.
Specific embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, below in conjunction with specific embodiment, to this
Invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, not
For limiting the present invention.
The present invention provides a kind of preparation methods of bacillus amyloliquefaciens G5 gemma bacterium powders, include the following steps：
The first step prepares production gemma culture medium
Producing gemma culture medium raw material is：Carbon source, nitrogen source and inorganic salts；
The production gemma culture medium each group is divided into：10~40g/L of cornstarch, 0.5~3g/L of glucose, dregs of beans 5~
25g/L, 0.1~1.5g/L of calcium carbonate, 0.1~1.0g/L of ammonium chloride, 0.1~1.0g/L of dipotassium hydrogen phosphate, magnesium sulfate 0.01~
0.01~0.7g/L of 0.7g/L and manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank, water is added to adjust concentration；It is 7.5~7.8 to adjust pH again,
Then the high-temperature sterilization under the conditions of 121~123 DEG C；Obtain production gemma culture medium；
Second step culture gemma zymotic fluid
Under aseptic condition, the seed liquor of bacillus amyloliquefaciens G5 is transferred in the fermentation tank with volume ratio 5~6%,
The fermented and cultured in the production gemma culture medium；After fermentation, the fermentation tank is warming up to 50~60 DEG C, maintain 20~
30min obtains the gemma zymotic fluid of the bacillus amyloliquefaciens G5；
Sampling calculates total bacteria count after fermentation is completed；After obtaining the gemma zymotic fluid, sampling calculates gemma number and bud
Spore rate；
Third step prepares gemma bacterium powder
Ammonium sulfate and diatomite are added into the gemma zymotic fluid, is stirred evenly；Then plate-frame filtering is carried out, collects filter
Cake；The filter cake is dried with boiling drying equipment again, 25~40min is dried under the conditions of 70~80 DEG C；Finally crushed with pulverizer
The filter cake after drying crosses 40~60 mesh sieve；Obtain the gemma bacterium powder of bacillus amyloliquefaciens G5；In the gemma bacterium powder
Sampling calculates living bacteria count and effective viable bacteria yield.
The bacillus amyloliquefaciens G5 is the dominant strain isolated and purified in spoil fermented sample.
The seed liquor preparation process of the bacillus amyloliquefaciens G5 includes：
Step 1 actication of culture
Under aseptic condition, the bacillus amyloliquefaciens G5 is seeded to activation culture in beef-protein medium,
22~28h is cultivated under the conditions of 36~38 DEG C；It crosses and is forwarded on beef extract-peptone tablet again, separation single bacterium colony, 36~38 DEG C
Under the conditions of cultivate 22~28h, repeatedly activate 2~3 times；
Step 2 prepares seed liquor
Under aseptic condition, single bacterium colony described in picking is forwarded in the beef extract-peptone fluid nutrient medium of sterilizing, 36~38
8~12h is cultivated under the conditions of DEG C, obtains the seed liquor.
The carbon source raw material of the production gemma culture medium further includes：Maize flour and sucrose；Nitrogen source raw material further includes：Fish meal and egg
White peptone；Inorganic salt raw material further includes：Ammonium sulfate, potassium dihydrogen phosphate and ferric trichloride.
The condition of culture of the fermentation tank is：23~36h is cultivated under the conditions of 36~38 DEG C, rotating speed is 180~200rpm, is led to
Air quantity is 1：1.2~1.5, tank pressure is 0.05~0.08Mpa.
As a preferred solution, the charge of the fermentation tank is 60~70%.
As a preferred solution, the ammonium sulfate and the diatomite account for the proportion difference of the gemma zymotic fluid
It is：Ammonium sulfate is 30%~40%, and diatomite is 3%~8%；Using ammonium sulfate as flocculant, using diatomite as filter aid,
The time of plate-frame filtering can be reduced, improves living bacteria count amount and effective viable bacteria yield in the gemma bacterium powder.
Gemma rate >=98% of the gemma zymotic fluid.
Effective viable bacteria yield >=95% in the gemma bacterium powder.
For acquisition preferred forms, only change the concentration that gemma culture medium each component is produced described in the above-mentioned first step, if
Put following several embodiments.
Following embodiment only lists the part different from above-mentioned steps, and identical content is no longer listed.
Embodiment one
The first step prepares production gemma culture medium
Producing gemma nutrient media components is：Carbon source, nitrogen source and inorganic salts；The carbon source raw material is cornstarch and glucose；
The nitrogen source raw material is dregs of beans；The inorganic salt raw material is calcium carbonate, ammonium chloride, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank；Water is added to adjust the production gemma culture medium each component dense
It spends and is：Cornstarch 15g/L, glucose 1g/L, dregs of beans 10g/L, calcium carbonate 0.4g/L, ammonium chloride 0.2g/L, dipotassium hydrogen phosphate
0.5g/L, magnesium sulfate 0.3g/L and manganese sulfate 0.2g/L；It is 7.5~7.8 to adjust pH again, then high under the conditions of 121~123 DEG C
Temperature sterilizing；Obtain production gemma culture medium.
After the production gemma medium culture bacillus amyloliquefaciens G5 of above-mentioned concentration, obtain the gemma number for 1.5 ×
1010CFU/ml, the gemma rate are 98.4%.
Embodiment two
The first step prepares production gemma culture medium
Producing gemma nutrient media components is：Carbon source, nitrogen source and inorganic salts；The carbon source raw material is cornstarch and glucose；
The nitrogen source raw material is dregs of beans；The inorganic salt raw material is calcium carbonate, ammonium chloride, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank；Water is added to adjust the production gemma culture medium each component dense
It spends and is：Cornstarch 20g/L, glucose 2.3g/L, dregs of beans 8g/L, calcium carbonate 0.5g/L, ammonium chloride 0.8g/L, dipotassium hydrogen phosphate
0.4g/L, magnesium sulfate 0.1g/L and manganese sulfate 0.2g/L；It is 7.5~7.8 to adjust pH again, then high under the conditions of 121~123 DEG C
Temperature sterilizing；Obtain production gemma culture medium.
After the production gemma medium culture bacillus amyloliquefaciens G5 of above-mentioned concentration, obtain the gemma number for 3.2 ×
1010CFU/ml, the gemma rate are 98.6%.
Embodiment three
The first step prepares production gemma culture medium
Producing gemma nutrient media components is：Carbon source, nitrogen source and inorganic salts；The carbon source raw material is cornstarch and glucose；
The nitrogen source raw material is dregs of beans；The inorganic salt raw material is calcium carbonate, ammonium chloride, dipotassium hydrogen phosphate, magnesium sulfate and manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank；Water is added to adjust the production gemma culture medium each component dense
It spends and is：Cornstarch 33g/L, glucose 1.4g/L, dregs of beans 22g/L, calcium carbonate 1.2g/L, ammonium chloride 0.7g/L, phosphoric acid hydrogen two
Potassium 0.6g/L, magnesium sulfate 0.5g/L and manganese sulfate 0.5g/L；It is 7.5~7.8 to adjust pH again, then under the conditions of 121~123 DEG C
High-temperature sterilization；Obtain production gemma culture medium.
After the production gemma medium culture bacillus amyloliquefaciens G5 of above-mentioned concentration, obtain the gemma number for 3.3 ×
1010CFU/ml, the gemma rate are 98.5%.
The implementation procedure of other embodiments repeats no more；Other concentration of each embodiment gemma culture medium each component and yield are shown in
Table one.
One gemma culture medium each component concentration of table and yield
Found out by data in above-mentioned table, the production gemma culture medium each component concentration range is：10~40g/ of cornstarch
L, 0.5~3g/L of glucose, 5~25g/L of dregs of beans, 0.1~1.5g/L of calcium carbonate, 0.1~1.0g/L of ammonium chloride, dipotassium hydrogen phosphate
0.01~0.7g/L of 0.1~1.0g/L, 0.01~0.7g/L of magnesium sulfate and manganese sulfate；Obtain the gemma number of the gemma zymotic fluid
It is 1.5 × 1010~3.5 × 1010CFU/ml, gemma rate >=98%.
Optional one group from the gemma culture medium of one various concentration of table, with a kind of above-mentioned bacillus amyloliquefaciens G5 Bacillus
The preparation method of powder only changes the proportion of flocculant described in third step, sets following several embodiments.
Following embodiment only lists the part different from above-mentioned preparation method step, and identical content is no longer listed.
Example IV
Third step prepares gemma bacterium powder
Flocculant is added into the gemma zymotic fluid, the flocculant proportion is：Ammonium sulfate is 34%, diatomite is
6%, it stirs evenly；Then plate-frame filtering is carried out, collects filter cake；Again the filter cake is dried with boiling drying equipment, 70~80 DEG C
Under the conditions of dry 25~40min；The filter cake after drying finally is crushed with pulverizer, crosses 40~60 mesh sieve；Obtain solution starch
The gemma bacterium powder of bacillus G5.
Bacillus amyloliquefaciens G5 gemma bacterium powders are prepared with the flocculant of above-mentioned proportion, are obtained in the gemma bacterium powder effectively
Viable count is 2.4 × 1011CFU/ml, the yield of effective viable bacteria is 95.7%.
Embodiment five
Third step prepares gemma bacterium powder
Flocculant is added into the gemma zymotic fluid, the flocculant proportion is：Ammonium sulfate is 36%, diatomite is
5%, it stirs evenly；Then plate-frame filtering is carried out, collects filter cake；Again the filter cake is dried with boiling drying equipment, 70~80 DEG C
Under the conditions of dry 25~40min；The filter cake after drying finally is crushed with pulverizer, crosses 40~60 mesh sieve；Obtain solution starch
The gemma bacterium powder of bacillus G5.
Bacillus amyloliquefaciens G5 gemma bacterium powders are prepared with the flocculant of above-mentioned proportion, are obtained in the gemma bacterium powder effectively
Viable count is 2.3 × 1011CFU/ml, the yield of effective viable bacteria is 95.6%.
Embodiment six
Third step prepares gemma bacterium powder
Flocculant is added into the gemma zymotic fluid, the flocculant proportion is：Ammonium sulfate is 35%, diatomite is
5%, it stirs evenly；Then plate-frame filtering is carried out, collects filter cake；Again the filter cake is dried with boiling drying equipment, 70~80 DEG C
Under the conditions of dry 25~40min；The filter cake after drying finally is crushed with pulverizer, crosses 40~60 mesh sieve；Obtain solution starch
The gemma bacterium powder of bacillus G5.
Bacillus amyloliquefaciens G5 gemma bacterium powders are prepared with the flocculant of above-mentioned proportion, are obtained in the gemma bacterium powder effectively
Viable count is 2.4 × 1011CFU/ml, the yield of effective viable bacteria is 95.7%.
The implementation procedure of other embodiments repeats no more；Each other proportions of embodiment flocculant and yield are shown in Table two.
Two flocculant proportion of table and yield
| Ammonium sulfate % | Diatomite % | Living bacteria count CFU/ml | Effective viable bacteria yield % | |
| 1 | 35 | 5 | 2.3×1011 | 95.4 |
| 2 | 37 | 4 | 2.2×1011 | 95.6 |
| 3 | 34 | 6 | 2.4×1011 | 95.7 |
| 4 | 36 | 5 | 2.3×1011 | 95.6 |
| 5 | 39 | 3 | 2.2×1011 | 95.2 |
| 6 | 31 | 8 | 2.2×1011 | 95.5 |
| 7 | 34 | 7 | 2.3×1011 | 95.5 |
| 8 | 35 | 5 | 2.4×1011 | 95.7 |
| 9 | 38 | 4 | 2.1×1011 | 95.6 |
| 10 | 36 | 3 | 2.1×1011 | 95.0 |
Found out by data in above-mentioned table, the flocculant proportion is：Ammonium sulfate is 30%~40%, diatomite be 3%~
8%；It is 2.2 × 10 to obtain living bacteria count in the gemma bacterium powder11~2.4 × 1011CFU/ml, the yield of effective viable bacteria >=
95%.
In conclusion the purpose of the present invention is to provide a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders,
By providing a kind of novel production gemma culture medium, while cultivate on novel production gemma culture medium the bud of bacillus amyloliquefaciens G5
Spore zymotic fluid, then using ammonium sulfate and diatomite as flocculant, then carry out plate-frame filtering, obtain bacillus amyloliquefaciens G5 buds
Spore bacterium powder.Present invention production gemma culture medium raw material is relatively common, is conveniently easy to get, greatly reduces production cost；In fermentation process,
Zymotechnique is simple, easy to control, does not need to carry out feed supplement；Number of spores in obtained bacillus amyloliquefaciens G5 gemma zymotic fluids
It is more, gemma rate >=98%；Meanwhile using ammonium sulfate and diatomite as flocculant, shorten the time prepared by bacterium powder, obtained Xie Dian
Living bacteria count is 4~6 times higher than the prior art in afnyloliquefaciens G5 gemma bacterium powders, yield >=95% of effective viable bacteria.
Certainly, the present invention can also have other various embodiments, without deviating from the spirit and substance of the present invention, ripe
It knows those skilled in the art and makes various corresponding changes and deformation, but these corresponding changes and change in accordance with the present invention
Shape should all belong to the protection domain of appended claims of the invention.
Claims (9)
1. a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders, which is characterized in that include the following steps：
The first step prepares production gemma culture medium
Producing gemma culture medium raw material is：Carbon source, nitrogen source and inorganic salts；
The production gemma culture medium each group is divided into：10~40g/L of cornstarch, 0.5~3g/L of glucose, 5~25g/L of dregs of beans,
0.1~1.5g/L of calcium carbonate, 0.1~1.0g/L of ammonium chloride, 0.1~1.0g/L of dipotassium hydrogen phosphate, 0.01~0.7g/L of magnesium sulfate
With 0.01~0.7g/L of manganese sulfate；
The production gemma culture medium each component is poured into fermentation tank, water is added to adjust concentration；It is 7.5~7.8 to adjust pH again, then
The high-temperature sterilization under the conditions of 121~123 DEG C；Obtain production gemma culture medium；
Second step culture gemma zymotic fluid
Under aseptic condition, the seed liquor of bacillus amyloliquefaciens G5 is transferred in the fermentation tank with volume ratio 5~6%, in institute
State fermented and cultured in production gemma culture medium；After fermentation, the fermentation tank is warming up to 50~60 DEG C, maintains 20~30min,
Obtain the gemma zymotic fluid of the bacillus amyloliquefaciens G5；
Third step prepares gemma bacterium powder
Ammonium sulfate and diatomite are added into the gemma zymotic fluid, is stirred evenly；Then plate-frame filtering is carried out, collects filter cake；
The filter cake is dried with boiling drying equipment again, 25~40min is dried under the conditions of 70~80 DEG C；It is finally crushed and dried with pulverizer
The filter cake afterwards crosses 40~60 mesh sieve；Obtain the gemma bacterium powder of bacillus amyloliquefaciens G5.
A kind of 2. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
It is the dominant strain isolated and purified in spoil fermented sample to state bacillus amyloliquefaciens G5.
A kind of 3. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
The seed liquor preparation process for stating bacillus amyloliquefaciens G5 includes：
Step 1 actication of culture
Under aseptic condition, the bacillus amyloliquefaciens G5 is seeded to activation culture in beef-protein medium, 36~
22~28h is cultivated under the conditions of 38 DEG C；It crosses and is forwarded on beef extract-peptone tablet again, detach single bacterium colony, 36~38 DEG C of conditions
22~28h of lower culture is activated 2~3 times repeatedly；
Step 2 prepares seed liquor
Under aseptic condition, single bacterium colony described in picking is forwarded in the beef extract-peptone fluid nutrient medium of sterilizing, 36~38 DEG C of items
8~12h is cultivated under part, obtains the seed liquor.
A kind of 4. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
The carbon source raw material for stating production gemma culture medium further includes：Maize flour and sucrose；Nitrogen source raw material further includes：Fish meal and peptone；It is inorganic
Salt raw material further includes：Ammonium sulfate, potassium dihydrogen phosphate and ferric trichloride.
A kind of 5. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
The condition of culture for stating fermentation tank is：Under the conditions of 36~38 DEG C cultivate 23~36h, rotating speed be 180~200rpm, ventilation quantity 1：
1.2~1.5, tank pressure is 0.05~0.08Mpa.
A kind of 6. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
The charge for stating fermentation tank is 60~70%.
A kind of 7. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 1, which is characterized in that institute
It states ammonium sulfate and the diatomite accounts for the proportion of the gemma zymotic fluid and is respectively：Ammonium sulfate is 30%~40%, and diatomite is
3%~8%.
8. a kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 3~7 any one,
It is characterized in that, gemma rate >=98% in the gemma zymotic fluid.
A kind of 9. preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders according to claim 8, which is characterized in that institute
State effective viable bacteria yield >=95% in gemma bacterium powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810095745.9A CN108148783A (en) | 2018-01-31 | 2018-01-31 | A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810095745.9A CN108148783A (en) | 2018-01-31 | 2018-01-31 | A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108148783A true CN108148783A (en) | 2018-06-12 |
Family
ID=62459134
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810095745.9A Pending CN108148783A (en) | 2018-01-31 | 2018-01-31 | A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108148783A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109468259A (en) * | 2018-10-24 | 2019-03-15 | 亚太星原农牧科技海安有限公司 | A kind of culture medium for promoting gemma to generate |
| CN110878274A (en) * | 2019-12-30 | 2020-03-13 | 中化农业生态科技(湖北)有限公司 | Bacillus amyloliquefaciens culture process method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102719383A (en) * | 2012-06-29 | 2012-10-10 | 武汉工业学院 | Bacillus amyloliquefaciens, inoculant and application thereof |
| CN103131655A (en) * | 2013-03-06 | 2013-06-05 | 江苏苏滨生物农化有限公司 | Bacillus amyloliquefaciens K-8 and bactericide thereof |
| CN103814892A (en) * | 2014-03-11 | 2014-05-28 | 江苏省苏科农化有限责任公司 | Bacillus amyloliquefaciens water dispersible granule |
| US20140352376A1 (en) * | 2013-05-28 | 2014-12-04 | BiOWiSH Technologies, Inc. | Fertilizer compositions methods of making and using same |
-
2018
- 2018-01-31 CN CN201810095745.9A patent/CN108148783A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102719383A (en) * | 2012-06-29 | 2012-10-10 | 武汉工业学院 | Bacillus amyloliquefaciens, inoculant and application thereof |
| CN103131655A (en) * | 2013-03-06 | 2013-06-05 | 江苏苏滨生物农化有限公司 | Bacillus amyloliquefaciens K-8 and bactericide thereof |
| US20140352376A1 (en) * | 2013-05-28 | 2014-12-04 | BiOWiSH Technologies, Inc. | Fertilizer compositions methods of making and using same |
| CN103814892A (en) * | 2014-03-11 | 2014-05-28 | 江苏省苏科农化有限责任公司 | Bacillus amyloliquefaciens water dispersible granule |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109468259A (en) * | 2018-10-24 | 2019-03-15 | 亚太星原农牧科技海安有限公司 | A kind of culture medium for promoting gemma to generate |
| CN109468259B (en) * | 2018-10-24 | 2022-04-05 | 亚太星原农牧科技海安有限公司 | Culture medium for promoting spore generation |
| CN110878274A (en) * | 2019-12-30 | 2020-03-13 | 中化农业生态科技(湖北)有限公司 | Bacillus amyloliquefaciens culture process method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106479927B (en) | Utilize the method and its application of bacillus licheniformis biosynthesis nanometer selenium | |
| CN102725397B (en) | Dead lactobacillus biomass for antimicrobial use and a production method therefor | |
| CN104106370B (en) | A kind of edible fungi liquid strain and production method thereof | |
| CN103190538B (en) | Aflatoxin B1 degradation agent and application thereof | |
| CN109306329B (en) | Bacillus subtilis and application thereof | |
| CN103146617A (en) | Bacillus subtilis and culture method and application thereof | |
| CN103396956B (en) | A kind of saccharomyces cerevisiae and its screening and culturing method and the method for bean pulp fermentation | |
| CN105925497A (en) | Bacillus pumilus and application thereof in decomposing phosphate and potassium and producing acid | |
| CN105132312A (en) | Bacillus subtilis and application thereof as well as microbial fungicide containing bacillus subtilis and preparation method of microbial fungicide | |
| CN105123763A (en) | Applications of bacillus subtilis fermentation broth, microbial fungicide containing bacillus subtilis, and preparation method of microbial fungicide | |
| CN106011001A (en) | Livestock intestinal ecological restoration microbial inoculum with antiviral and antibacterial effects and capable of replacing antibiotics and preparation method of livestock intestinal ecological restoration microbial inoculum | |
| CN110358710A (en) | One plant of bacillus laterosporus and preparing the application in disease-resistant saline-alkali tolerant functional microorganism preparation | |
| CN105820986B (en) | The special rhizosphere growth-promoting A Shi bacillus of one plant of blueberry and its application | |
| CN101851121B (en) | Compound bacterial agent for efficiently converting pig excrements and preparation method and application thereof | |
| CN113549574A (en) | Bacillus coagulans and application thereof | |
| CN106754371A (en) | A kind of microbial bacterial agent preparation method and application for improving soil phosphorus utilization | |
| CN108148783A (en) | A kind of preparation method of bacillus amyloliquefaciens G5 gemma bacterium powders | |
| CN105439726A (en) | Bacillus megaterium bacterial fertilizer for onsite fermentation in farm and applications thereof | |
| CN110684693A (en) | Fermentation method of bacillus subtilis and preparation method of microbial inoculum thereof | |
| CN102719363B (en) | Preparation method of antibacterial fermentation liquid of Solidago canadesis endophytic fungi | |
| CN106635919A (en) | Spirulina culture method | |
| CN111733117B (en) | Bacillus marinus for producing antibacterial peptide and fermentation method and application thereof | |
| CN104757544B (en) | A kind of dietary supplements with bacteriostasis and its preparation method and application | |
| CN116622547A (en) | Bacillus mojavensis YL-78 and application thereof | |
| CN116024114A (en) | Intestinal bacillus subtilis strain and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180612 |
|
| RJ01 | Rejection of invention patent application after publication |