CN101032527A - Active clostridium butyrium agent and the preparing method thereof - Google Patents
Active clostridium butyrium agent and the preparing method thereof Download PDFInfo
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- CN101032527A CN101032527A CNA2006100444347A CN200610044434A CN101032527A CN 101032527 A CN101032527 A CN 101032527A CN A2006100444347 A CNA2006100444347 A CN A2006100444347A CN 200610044434 A CN200610044434 A CN 200610044434A CN 101032527 A CN101032527 A CN 101032527A
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- 241000193403 Clostridium Species 0.000 title claims description 22
- 238000000034 method Methods 0.000 title claims description 12
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims abstract description 114
- 241000894006 Bacteria Species 0.000 claims abstract description 73
- 239000001963 growth medium Substances 0.000 claims abstract description 34
- 238000011218 seed culture Methods 0.000 claims abstract description 29
- 238000000855 fermentation Methods 0.000 claims abstract description 28
- 230000004151 fermentation Effects 0.000 claims abstract description 27
- 238000002360 preparation method Methods 0.000 claims abstract description 16
- 238000001035 drying Methods 0.000 claims abstract description 10
- 239000000843 powder Substances 0.000 claims abstract description 9
- 238000012856 packing Methods 0.000 claims abstract description 7
- 241001465754 Metazoa Species 0.000 claims abstract description 6
- 239000000203 mixture Substances 0.000 claims abstract description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 20
- 239000003795 chemical substances by application Substances 0.000 claims description 20
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- 239000002609 medium Substances 0.000 claims description 15
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- 239000001888 Peptone Substances 0.000 claims description 10
- 108010080698 Peptones Proteins 0.000 claims description 10
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 10
- 238000004108 freeze drying Methods 0.000 claims description 10
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 10
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 10
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 10
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 10
- 239000002994 raw material Substances 0.000 claims description 10
- 239000012138 yeast extract Substances 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 9
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- 239000002054 inoculum Substances 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 229920002472 Starch Polymers 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
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- 239000010931 gold Substances 0.000 claims 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses one kind of live butyric acid bacteria preparation and its preparation process. The live butyric acid bacteria preparation is prepared through three stage seed culture of butyric acid bacteria in the seed culture medium, fermentation in the butyric acid bacteria fermenting culture medium, collecting thallus, drying to obtain butyric acid bacteria powder, adding supplementary material, packing and other steps. The composition of the seed culture medium and that of the butyric acid bacteria fermenting culture medium are also disclosed. The live butyric acid bacteria preparation has high spore rate, high live butyric acid bacteria content and low cost, and may be used for preventing and treating diseases of both human body and animal, including human gastrointestinal tract disease, hog and chicken white flux, etc.
Description
Technical field the invention belongs to the microorganism formulation technical field, relates to a kind of active clostridium butyrium agent and preparation method thereof.
The background technology butyric acid bacteria is that Clostridium butyricum (Clostridium butyrium) is the Gram-positive anaerobic spore-bearing bacilli that belongs to fusobacterium, is present in the intestinal of soil, animal and human's body.Its main metabolites is butanoic acid, acetic acid, and wherein butanoic acid is the indispensable energy of animal body gut epithelium histiocyte metabolism, and the reparation of intestinal epithelial tissue due to the intestinal illness is had a very important role; Butyric acid bacteria is one of normal flora in the humans and animals body, it can promote the propagation and the growth of profitable strain (bacillus bifidus, lactobacillus etc.) in the body intestinal, suppress the growth and breeding of the interior putrefaction bacteria of intestinal and harmful bacterium, correct flora disorder in the intestinal, reduce and reduce the generation of enterotoxin; Different with intestinal beneficial bacteriums such as bacillus bifidus, lactobacilluss is, because butyric acid bacteria has spore, thereby environment has stronger resistance to external world, it be heated to 90 ℃ 10 minutes, 100 ℃ 5 minutes can inactivation, still can survive in the environment of PH 1.0-5.0, can be fit to its development growth in the environment of PH 4.0-9.8, its solid live bacteria preparation is preserved under room temperature, drying regime has good stable, preserve more than 2 years, do not see that viable bacteria significantly reduces.
Have its unique biological characteristics just because of butyric acid bacteria, as far back as the forties, Japan just is developed to it the medicine of treatment gastroenteropathy.Nearly half a century, the active clostridium butyrium agent that Japan produces---the refined series of products of rice are in Asian countries such as Japan, Korea S, China and regionally be widely used in the treatment of diseases such as alteration of intestinal flora, acute and chronic diarrhea, irritable bowel syndrome, antibiotic dependency enteritis, constipation or alternating diarrhea and constipation disease that various reasons such as digestive system, children's, surgery, tumor, department of obstetrics and gynecology cause, determined curative effect, in addition, the butyric acid bacteria preparation also can be used as enteritis, various hepatopathy, chemoradiotherapy and causes that body immunity descends, the good adjuvant therapy medicaments of dysbacteriosis.
Before this century, China does not have the active clostridium butyrium agent product oneself produced, and Chinese various big hospital is the japanese product of using clinically always.Over the past two years, domestic priority has two tame enterprises to obtain the New Drug Certificate of active clostridium butyrium agent product, and realized the industrialization of this product, but it is less than normal also to exist production scale, lyophilization mycopowder viable bacteria content is high not enough, the spore formation rate is on the low side, production cost height, problems such as product shelf stability difference.But, do not find the production and the application of active clostridium butyrium agent for animals at present as yet.
Summary of the invention the purpose of this invention is to provide the active clostridium butyrium agent of a kind of high spore formation rate, high viable bacteria content.
Another object of the present invention provides a kind of preparation method of active clostridium butyrium agent
For achieving the above object, the present invention has taked following technical scheme:
Active clostridium butyrium agent of the present invention, by butyric acid bacteria in the butyric acid bacteria seed culture medium through three grades of seed amplification culture with in the butyric acid bacteria fermentation medium, ferment, make through microorganism collection, dry powder process, mycopowder check, batching, product inspection, packaging process again;
Said butyric acid bacteria seed culture medium is made up of following raw material and weight proportion:
Peptone 0.5%～2.0%, yeast extract 0.3%～1.8%, dipotassium hydrogen phosphate 0.02%～0.5%, potassium dihydrogen phosphate 0.02%～0.5%, magnesium sulfate 0.02%～0.2%, glucose 0.8%～2.0%, water 94.5%～96.8%, the PH 5.0～8.5 of culture medium;
Said butyric acid bacteria fermentation medium is made up of following raw material and weight proportion:
Peptone 0.5%～2.0%, yeast extract 0.3%～1.8%, dipotassium hydrogen phosphate 0.02%～0.5%, potassium dihydrogen phosphate 0.02%～0.5%, magnesium sulfate 0.02%～0.2%, starch 0.2%～2.0%, calcium carbonate 0.1%～1.0%, water 93.8%～97.0%, the PH 5.0～8.5 of culture medium.
The preparation method of active clostridium butyrium agent of the present invention is made up of following processing step successively:
1, first order seed is cultivated: get butyric acid bacteria seeding lyophilizing pipe strain, be inoculated in the butyric acid bacteria seed culture medium, cultivate 14～18h 30～38 ℃ of anaerobism;
2, secondary seed is cultivated: first order seed is inoculated in the butyric acid bacteria seed culture medium with 5～10% inoculum concentration, cultivates 14～18h 30～38 ℃ of anaerobism;
3, three grades of seed culture: secondary seed is inoculated in the butyric acid bacteria seed culture medium with 5～10% inoculum concentration, cultivates 13～18h 30～38 ℃ of anaerobism, and seed culture at different levels finish and all should check its growth conditions and purity;
4, fermentation: three grades of seeds are inoculated in the butyric acid bacteria fermentation medium with 5～10% inoculum concentration, control tank pressure 0.04MPa～0.06MPa cultivates 16～24h 30～38 ℃ of anaerobic fermentations, until spore formation rate＞80%, fermentation finishes, and should to check that it has pollution-free, should give discarded if any pollution;
5, microorganism collection: the zymocyte liquid through being up to the standards, adopt the high speed tube centrifuge to collect thalline, rotating speed 〉=16000r/min obtains wet bacterium mud;
6, dry powder process: wet bacterium mud is weighed, weight ratio by 1: 2～10, the adding weight ratio is 20% aseptic defat milk, mix homogeneously carries out vacuum lyophilization, earlier at-40 ℃ of pre-freeze 2～3h, evacuation then, vacuum is 2～20Pa, and temperature rises to 28 ℃ gradually in the dry run, and whole dry run is 20～28h.Pulverize after drying finishes, cross 80 mesh sieves, make mycopowder;
7, mycopowder check: every gram mycopowder contains the butyric acid bacteria viable count and is no less than 1.0 * 10
9Individual;
8, batching: the prior mycopowder of making in corn starch and the step 6 of 120 ℃ of drying 4～6h of general 9～50: 1 ratio by weight ratio carries out batch mixes, promptly gets the finished product of active clostridium butyrium agent of the present invention;
9, product inspection: require every gram finished product to contain the butyric acid bacteria viable count and be no less than 1.0 * 10
8Individual;
10, packing is dispatched from the factory: packing can be dispatched from the factory after qualified after sampling is examined entirely.
The optimum PH of described culture medium is 6.5～7.5, and the suitableeest fermentation temperature is 36～37 ℃.
The active clostridium butyrium agent of the present invention's preparation, spore rate height, the viable bacteria content height, cheap, the people beast is dual-purpose, can be used for the human body intestinal canal flora imbalance that a variety of causes causes, acute and chronic diarrhea, irritable bowel syndrome, with antibiotic dependency enteritis, treatment of diseases such as constipation or alternating diarrhea and constipation disease, and pass through pig, the toxicity of chicken, pharmacology and pharmacodynamic study and large-scale clinical trial prove: add 3.0 ‰ active clostridium butyrium agent in feedstuff, to pig, chicken has no adverse effects, the piglet diarrhea sickness rate has reduced by 36.9%, and prevention and growth promoting function are preferably arranged; Treat piglet, DISEASE IN FLOCKS with active clostridium butyrium agent, curative effect is preferably all arranged, average cure rate is respectively 80.64% and 95.91%, has met or exceeded the average cure rate of antibiotics; In feedstuff, add 0.5 ‰～1.0 ‰ active clostridium butyrium agent, can effectively prevent the intestinal tract disease of chicken, pig, and promote their growth effectively.
The specific embodiment is an example with 500 liters of fermentation tanks
Embodiment 1
The butyric acid bacteria seed culture medium is made up of following raw material and weight proportion:
Peptone 0.5, yeast extract 1.8, dipotassium hydrogen phosphate 0.5, potassium dihydrogen phosphate 0.5, magnesium sulfate 0.2, glucose 2.0, water, 94.5 (water is by ml, and surplus person is by g), the PH5.0 of seed culture～6.0;
The butyric acid bacteria fermentation medium is made up of following raw material and weight proportion:
Peptone 0.5, yeast extract 1.8, dipotassium hydrogen phosphate 0.5, potassium dihydrogen phosphate 0.5, magnesium sulfate 0.2, starch 2.0, calcium carbonate 1.0, water 93.5 (water is by ml, and surplus person is by g), the PH5.0 of fermentation medium～6.0;
Butyric acid bacteria cryovial strain is provided by the Jiangsu Institute of Microbiology Co., Ltd
Finish by following processing step:
1, the cultivation of first order seed: seed culture medium 100ml is housed in the 150ml triangular flask, behind 121 ℃ of sterilization 30min, inserts butyric acid bacteria cryovial strain, cultivate down in anaerobic condition, 30～31 ℃ of temperature are cultivated 14h.
2, secondary seed is cultivated: seed culture medium 1000ml is housed in the 2000ml triangular flask, inserts long good first order seed 50ml behind 121 ℃ of sterilization 30min, cultivate down in anaerobic condition, 30～31 ℃ of temperature are cultivated 14h.
3, three grades of seed culture: 19 liters of culture medium are housed in 50 liters of jars, and 121 ℃ of sterilization 30min insert 1 liter of long good secondary seed, cultivate down in anaerobic condition, and 30～31 ℃ of temperature are cultivated 13～15h.
4,500 liters of fermentor cultivation: with 380 liters of fermentation medium loading amounts, 121 ℃ of sterilization 30min insert 20 liters in long three grades of good seeds, control tank pressure 0.04MPa～0.06MPa, 30～31 ℃ of fermentation temperatures, incubation time 16～24h.
5, microorganism collection: cultured fermentation liquid is collected thalline with tube centrifuge, and rotating speed 〉=16000r/min obtains wet bacterium mud.
6, dry powder process: will wet bacterium mud and weight ratio are 20% aseptic defatted milk by 1: 2 weight ratio mixing, pack into and carry out lyophilization in the freezer dryer, earlier at-40 ℃ of pre-freeze 2～3h, evacuation then, vacuum is 2～20Pa, temperature rises to 28 ℃ gradually in the dry run, and whole dry run is 20～28h.Drying finishes, and dried truffle is pulverized, crossed 80 mesh sieves, makes mycopowder.Adopt above-mentioned technology can get lyophilization mycopowder 2.8～3.0Kg, mycopowder contains butyric acid bacteria viable count 〉=1.0 * 10
9Individual/gram.
7, finished product preparation: an amount of auxiliary materials and mixing of viable count adding according to mycopowder is made finished product, and every gram finished product contains the butyric acid bacteria viable count and is no less than 1.0 * 10
8Individual.Adjuvant is the corn starch of prior 120 ℃ of drying 4～6h, is packaged into the product of different size after mycopowder and adjuvant are fully mixed.
Embodiment 2 is according to embodiment 1, and the butyric acid bacteria seed culture medium is made up of following raw material and weight proportion:
Peptone 1.25, yeast extract 1.15, dipotassium hydrogen phosphate 0.26, potassium dihydrogen phosphate 0.26, magnesium sulfate 0.11, glucose 1.4, water 95.57 (water is by ml, and surplus person is by g), the PH6.5 of seed culture～7.5;
The butyric acid bacteria fermentation medium is made up of following raw material and weight proportion:
Peptone 1.25, yeast extract 1.25, dipotassium hydrogen phosphate 0.26, potassium dihydrogen phosphate 0.26, magnesium sulfate 0.11, starch 1.4, calcium carbonate 0.55, water 93.5 (water is by ml, and surplus person is by g), the PH6.5 of fermentation medium～7.5;
1, the cultivation of first order seed: the butyric acid bacteria cryovial is seeded to respectively in 2 150ml triangular flasks that the 110ml seed culture medium is housed, cultivates down, temperature 34-35 ℃, cultivate 16h in anaerobic condition.
2, the cultivation of secondary seed: dress culture medium 1390ml in 2 2000ml triangular flasks, insert long good first order seed 110ml behind 121 ℃ of sterilization 30min respectively, cultivate down in anaerobic condition, temperature 34-35 ℃, cultivate 16h.
3, the cultivation of three grades of seeds: the dress culture medium is 27.8 liters in 50 liters of jars, inserts 2.25 liters of long good secondary seeds behind 121 ℃ of sterilization 30min, cultivates down in anaerobic condition, temperature 34-35 ℃, cultivates 15～17h.
4,500 liters of fermentor cultivation: with 370 liters of above-mentioned fermentation medium loading amounts, 121 ℃ of sterilization 30min insert long 30 liters of good three grades of seeds, control tank pressure 0.04MPa～0.06MPa, 36～37 ℃ of fermentation temperatures, incubation time 16～24h.
5, microorganism collection: method is with embodiment 1.
6, dry powder process: the aseptic defatted milk of will wet bacterium mud and 20% carries out lyophilization in 1: 6 ratio mixing in the freezer dryer of packing into, vacuum is 2～20Pa, and temperature is-40 ℃～28 ℃, and whole dry run is 20～28h.Drying finishes, and dried truffle is pulverized, crossed 80 mesh sieves, makes mycopowder.Adopt above-mentioned technology can get lyophilization mycopowder 3.0～3.2Kg, mycopowder contains butyric acid bacteria viable count 〉=1.0 * 10
9Individual/gram.
7, finished product preparation: method is with embodiment 1.
Embodiment 3 is according to embodiment 1, and the butyric acid bacteria seed culture medium is made up of following raw material and weight proportion:
Peptone 2.0, yeast extract 0.3, dipotassium hydrogen phosphate 0.02, potassium dihydrogen phosphate 0.02, magnesium sulfate 0.02, glucose 0.4, water 96.84 (water is by ml, and surplus person is by g), the PH7.5 of seed culture medium～8.5;
The butyric acid bacteria fermentation medium is made up of following raw material and weight proportion:
Peptone, 2.0 yeast extracts, 0.3 dipotassium hydrogen phosphate, 0.02 potassium dihydrogen phosphate, 0.02 magnesium sulfate, 0.02 starch, 0.2 calcium carbonate, 0.1 water, 97.34 (water is by ml, and surplus person is by g), the PH7.5 of fermentation medium～8.5;
1, the cultivation of first order seed: the butyric acid bacteria cryovial is seeded to respectively in 2 250ml triangular flasks that the 200ml seed culture medium is housed, cultivates down in anaerobic condition, 37～38 ℃ of temperature are cultivated 16～18h.
2, the cultivation of secondary seed: in 2 2000ml triangular flasks, adorn culture medium 1800ml respectively, insert long good first order seed 200ml behind 121 ℃ of sterilization 30min respectively, cultivate down, temperature 37-38 ℃, cultivate 16～18h in anaerobic condition.
3, the cultivation of three grades of seeds: the dress culture medium is 36 liters in 50 liters of jars, inserts 4 liters of long good secondary seeds behind 121 ℃ of sterilization 30min, cultivates down in anaerobic condition, temperature 37-38 ℃, cultivates 16～18h.
4,500 liters of fermentor cultivation: with 360 liters of above-mentioned fermentation medium loading amounts, 121 ℃ of sterilization 30min insert long 40 liters of good three grades of seeds, control tank pressure 0.04MPa～0.06MPa, 37～38 ℃ of temperature, incubation time 16～24h.
5, microorganism collection: method is with embodiment 1.
6, dry powder process: the aseptic defatted milk of will wet bacterium mud and 20% carries out lyophilization in 1: 10 ratio mixing in the freezer dryer of packing into, vacuum is 2～20Pa, and temperature is-40 ℃～28 ℃, and whole dry run is 20～28h.Drying finishes, and dried truffle is pulverized, crossed 80 mesh sieves, makes mycopowder.Adopt above-mentioned technology can get lyophilization mycopowder 3.0～3.2Kg, mycopowder contains butyric acid bacteria viable count 〉=1.0 * 10
9Individual/gram.
7, finished product preparation: method is with embodiment 1.
The present invention compares with domestic like product, and strain and production technology are all inequality.Mainly have the following advantages: 1, butyric acid bacteria is that the Jiangsu Institute of Microbiology Co., Ltd is separated acquisition voluntarily from soil, and strain good stability, product more than 2 years, still can have higher viable bacteria content in room temperature preservation; 2, seed culture medium and fermentation culture based component are all fairly simple, and all are daily buying, the low prices of being easy to, and obtain every kilogram of butyric acid bacteria lyophilized powder and (contain butyric acid bacteria viable count 〉=1.0 * 10
9Individual/gram) cost of material is 200 yuan, and generally need 400～500 yuan approximately; 3, butyric acid bacteria lyophilization powder viable bacteria content height, viable count is 1.0 * 10
9～1.0 * 10
10Individual/gram, spore content height, spore number 〉=80%; 4, active clostridium butyrium agent not only can be used for treating human intestines and stomach's tract disease, and the Hakuri of pig, chicken is had prevention and therapeutical effect preferably, and can effectively promote their growth.This at home still first.
Claims (3)
1, a kind of active clostridium butyrium agent, it is characterized in that by butyric acid bacteria in the butyric acid bacteria seed culture medium through three grades of seed amplification culture with in the butyric acid bacteria fermentation medium, ferment, make through microorganism collection, dry powder process, mycopowder check, batching, product inspection, packaging process again;
Said butyric acid bacteria seed culture medium is made up of following raw material and weight proportion:
Peptone 0.5%～2.0%, yeast extract 0.3%～1.8%, dipotassium hydrogen phosphate 0.02%～0.5%, potassium dihydrogen phosphate 0.02%～0.5%, magnesium sulfate 0.02%～0.2%, glucose 0.8%～2.0%, water 94.5%～96.8%, the PH 5.0～8.5 of culture medium;
Said butyric acid bacteria fermentation medium is made up of following raw material and weight proportion:
Peptone 0.5%～2.0%, yeast extract 0.3%～1.8%, dipotassium hydrogen phosphate 0.02%～0.5%, potassium dihydrogen phosphate 0.02%～0.5%, magnesium sulfate 0.02%～0.2%, starch 0.2%～2.0%, calcium carbonate 0.1%～1.0%, water 93.8%～97.0%, the PH 5.0～8.5 of culture medium.
2, its feature of the preparation method of active clostridium butyrium agent according to claim 1 is made up of following processing step successively:
(1), first order seed is cultivated: get butyric acid bacteria seeding lyophilizing pipe strain, be inoculated in the butyric acid bacteria seed culture medium, cultivate 14～18h 30～38 ℃ of anaerobism;
(2), secondary seed is cultivated: first order seed is inoculated in the butyric acid bacteria seed culture medium with 5～10% inoculum concentration, cultivates 14～18h 30～38 ℃ of anaerobism;
(3), three grades of seed culture: secondary seed is inoculated in the butyric acid bacteria seed culture medium with 5～10% inoculum concentration, cultivates 13～18h 30～38 ℃ of anaerobism, and seed culture at different levels finish and all should check its growth conditions and purity;
(4), fermentation: three grades of seeds are inoculated in the butyric acid bacteria fermentation medium with 5～10% inoculum concentration, control tank pressure 0.04MPa～0.06MPa cultivates 16～24h 30～38 ℃ of anaerobism, until spore formation rate＞80%, fermentation finishes, and should to check that it has pollution-free, should give discarded if any pollution;
(5), microorganism collection: the zymocyte liquid through being up to the standards, employing high speed tube centrifuge collection thalline, rotating speed 〉=16000r/min, the wet bacterium mud of acquisition;
(6), dry powder process: wet bacterium mud is weighed, weight ratio by 1: 2～10, the adding weight ratio is 20% aseptic defat milk, mix homogeneously carries out vacuum lyophilization, earlier at-40 ℃ of pre-freeze 2～3h, evacuation then, vacuum is 2～20Pa, and temperature rises to 28 ℃ gradually in the dry run, and whole dry run is 20～28h.Pulverize after drying finishes, cross 80 mesh sieves, make mycopowder;
(7), mycopowder check: every gram mycopowder contains the butyric acid bacteria viable count and is no less than 1.0 * 10
9Individual;
(8), batching: the prior mycopowder of making in corn starch and the step (6) of 120 ℃ of drying 4～6h of general 9～50: 1 ratio by weight ratio carries out batch mixes, promptly gets the finished product of the present invention's active clostridium butyrium agent for animals;
(9), product inspection: require every gram finished product to contain the butyric acid bacteria viable count and be no less than 1.0 * 10
8Individual;
(10), packing dispatches from the factory: packing is after gold is carried out in sampling examines, and dispatches from the factory after qualified.
3, the preparation method of active clostridium butyrium agent according to claim 2, the optimum PH that it is characterized in that described culture medium is 6.5～7.5, the suitableeest fermentation temperature is 36～37 ℃.
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| CNA2006100444347A CN101032527A (en) | 2006-03-09 | 2006-03-09 | Active clostridium butyrium agent and the preparing method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102199590A (en) * | 2011-04-02 | 2011-09-28 | 江苏省苏微微生物研究有限公司 | Method for preparing butyric acid bacteria powder by microencapsulated propagation culture and application |
| CN102321552A (en) * | 2011-08-12 | 2012-01-18 | 北京金泰得生物科技股份有限公司 | Clostridium butyricum used for feeding, and application thereof |
| CN102337238A (en) * | 2011-09-28 | 2012-02-01 | 重庆泰平药业有限公司 | Method for culturing microbes through fermentation |
| CN102352329A (en) * | 2011-09-28 | 2012-02-15 | 重庆泰平药业有限公司 | Production method for microbial germ powder |
| CN102660473A (en) * | 2012-05-04 | 2012-09-12 | 湖北绿雪生物产业有限公司 | Method for producing clostridium butyricum preparation by using continuous fermentation method |
| CN105039229A (en) * | 2015-09-08 | 2015-11-11 | 河北旺达饲料兽药开发有限公司 | Microorganism complex microbial agent and preparation method and application thereof |
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- 2006-03-09 CN CNA2006100444347A patent/CN101032527A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102199590A (en) * | 2011-04-02 | 2011-09-28 | 江苏省苏微微生物研究有限公司 | Method for preparing butyric acid bacteria powder by microencapsulated propagation culture and application |
| CN102199590B (en) * | 2011-04-02 | 2012-07-11 | 江苏省苏微微生物研究有限公司 | Method for preparing butyric acid bacteria powder by microencapsulated propagation culture and application |
| CN102321552A (en) * | 2011-08-12 | 2012-01-18 | 北京金泰得生物科技股份有限公司 | Clostridium butyricum used for feeding, and application thereof |
| CN102321552B (en) * | 2011-08-12 | 2014-04-16 | 北京金泰得生物科技股份有限公司 | Clostridium butyricum used for feeding, and application thereof |
| CN102337238A (en) * | 2011-09-28 | 2012-02-01 | 重庆泰平药业有限公司 | Method for culturing microbes through fermentation |
| CN102352329A (en) * | 2011-09-28 | 2012-02-15 | 重庆泰平药业有限公司 | Production method for microbial germ powder |
| CN102660473A (en) * | 2012-05-04 | 2012-09-12 | 湖北绿雪生物产业有限公司 | Method for producing clostridium butyricum preparation by using continuous fermentation method |
| CN105039229A (en) * | 2015-09-08 | 2015-11-11 | 河北旺达饲料兽药开发有限公司 | Microorganism complex microbial agent and preparation method and application thereof |
| CN105039229B (en) * | 2015-09-08 | 2019-06-14 | 河北普德动物药业有限公司 | A kind of complex microbial inoculum and its preparation method and application |
| CN109082395A (en) * | 2017-11-15 | 2018-12-25 | 浙江惠嘉生物科技股份有限公司 | Miyarisan cultural method |
| CN109082395B (en) * | 2017-11-15 | 2019-08-30 | 浙江惠嘉生物科技股份有限公司 | Miyarisan cultural method |
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