CN102000343A - Protective agent of live bacillus subtilis preparation - Google Patents
Protective agent of live bacillus subtilis preparation Download PDFInfo
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- CN102000343A CN102000343A CN2009100703097A CN200910070309A CN102000343A CN 102000343 A CN102000343 A CN 102000343A CN 2009100703097 A CN2009100703097 A CN 2009100703097A CN 200910070309 A CN200910070309 A CN 200910070309A CN 102000343 A CN102000343 A CN 102000343A
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- bacillus subtilis
- protective agent
- vitamin
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- active bacteria
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Abstract
The invention discloses a protective agent of a live bacillus subtilis preparation, comprising the following components (W/V): 3-4.5% of lactose, 0.5-1% of sodium carboxymethylcellulose, 0.5-1% of potassium sorbate, 0.1-0.5% of vitamin E, 0.1-0.5% of vitamin C, and 0.1-0.2% of water quality lemon. The protective agent of the invention solves the problem that the bacillus subtilis preparation produced by the conventional method is difficult to be stored, and has the advantages of simple and convenient method, easy operation, suitability for the batch preparation of the bacillus subtilis preparation, and high application value.
Description
Technical field
The present invention relates to field of biological product, provide a kind of protective agent of bacillus subtilis active bacteria formulation specifically.
Background technology
Bacillus subtilis is an obligate aerobe, and it is very capable to take oxygen by force, but can not grow under anaerobic state.In the birds intestinal, though can find bacillus subtilis, be to count seldom, even some propagation, also can very fast metabolism.Therefore develop the bacillus subtilis goods, replenish the bacillus subtilis quantity of intestinal field planting in vivo,, improve immunity, promote healthy, significant for adjusting the intestinal microecology balance by the external source mode.
In recent years, bacillus subtilis goods industry development is swift and violent both at home and abroad, becomes one of the fastest industry of global evolution, occupies more and more important position, and product category is various.Yet quality problems such as many bacillus subtilis goods ubiquity viable bacteria contents of present listing are low, the viable bacteria preservation term is short.Especially liquid product.Adopt the method for freezing, though can the liquid viable bacteria preservation term of significant prolongation, it is higher to keep freezing expense, and transportation is inconvenient, and is strong excessively to the dependency at freezing center.Bacillus subtilis living bacterial liquid body preparation adopts the bacillus subtilis excellent species through liquid fermentation and culture, and concentrating and separating is miscible with the protective agent medium.Liquid preparation not only viable bacteria content height, preservation term is long, and preservation and convenient transportation, and the mechanization degree height is convenient to industrialized mass production; Both can be directly oral, can be used as feed additive again, have wide promotion prospect.
Therefore the protective agent of studying the bacillus subtilis active bacteria formulation highly significant in the survival rate that keeps bacillus subtilis to greatest extent, makes the bacillus subtilis active bacteria formulation be easy to long preservation and vigor does not reduce, and is applicable to the demand of suitability for industrialized production more.
Summary of the invention
The protective agent that the purpose of this invention is to provide a kind of bacillus subtilis active bacteria formulation.
The special-purpose protecting agent that is used for the bacillus subtilis active bacteria formulation provided by the present invention, (W/V) forms by following component: lactose 3-4.5%, sodium carboxymethyl cellulose 0.5-1%, potassium sorbate 0.5-1%, vitamin E 0.1-0.5%, vitamin C 0.1-0.5%, water quality Fructus Citri Limoniae 0.1-0.2%.
The protective agent of above-mentioned bacillus subtilis active bacteria formulation, preferred content (W/V) lactose 4.5%, sodium carboxymethyl cellulose 0.5%, potassium sorbate 0.5%, vitamin E 0.1%, vitamin C 0.1%, water quality Fructus Citri Limoniae 0.1%.
The protective agent of above-mentioned bacillus subtilis active bacteria formulation is characterized in that containing 1.0 * 10 in every 1ml protective agent
9-1.0 * 10
14Cfu.
Beneficial effect of the present invention is: contain the protectant bacillus subtilis active bacteria formulation of the present invention and preserve 12 months viable counts at normal temperatures and still remain on higher level, and under the environment of temperature higher (37 ℃), can keep 6 months effectively.And the present invention is easy and simple to handle, is fit to preparation bacillus subtilis active bacteria formulation in enormous quantities, has solved the problem that the bacillus subtilis goods room temperature of producing with conventional method is preserved difficulty, has very high using value.
The specific embodiment
Method among the following embodiment if no special instructions, is conventional method.
Embodiment 1: the preparation of bacillus subtilis active bacteria formulation
Bacillus subtilis strain activates bacterial classification inoculation on the beef extract-peptone solid medium.Put in the incubator then and cultivate,, cultivate 16h at 37 ℃;
The fermentation culture of bacillus subtilis joins the bacterium colony eluting of above-mentioned activation culture in the beef extract-peptone fluid medium with aseptic manipulation, and 37 ℃, 200 commentaries on classics/min cultivate 16h.
The preparation of bacillus subtilis active bacteria formulation imports centrifuge with the fermentation liquid of above-mentioned fermentation by pipeline, and is centrifugal with the high capacity centrifuge under the condition of low temperature (4 ℃), 6000rpm, centrifugal 15min.Collect mud shape thalline in the concentrator bowl, 40 ℃ of drying under reduced pressure get bacillus subtilis dry bacterium powder 10g, and the check viable count is 2.2 * 10
16Cfu/g.
Protective agent preparation: take by weighing lactose 45g, sodium carboxymethyl cellulose 5.0g, potassium sorbate 5.0g, vitamin e1 .0g, vitamin C 1.0g, water quality Fructus Citri Limoniae 1.0g, get an amount of beef extract-peptone fluid medium dissolving after, mix homogeneously is settled to 1000ml.
Add protective agent and get above-mentioned protective agent solution 500ml, add above-mentioned bacillus subtilis dry bacterium powder 4.55g, stir, and be settled to 1000ml with protective agent solution.
Embodiment 2: the Detection of Stability of bacillus subtilis active bacteria formulation
The bacillus subtilis active bacteria formulation that will prepare according to the method for embodiment 1 is divided into 2 parts, be positioned over room temperature and 37 ℃ of casees that keep sample, the sample that room temperature is placed sampling in per 3 months detects viable count and viable bacteria index, and sample sampling every month of 37 ℃ of placements detects a viable count.
The viable count detection method:
1. the shake well fluid sample is drawn the 0.5ml fluid sample in test tube, adds the 4.5ml diluent, shake well, existing 1: 10 test liquid;
2. dilution: draw the 0.5ml test liquid, add the 4.5ml diluent in another test tube, fully mixing makes into 1: 100 test liquid, repeats aforesaid operations, carries out 10 times of serial dilutions, until required dilution factor;
3. drip sample: the bacterium liquid 0.2ml that draws the dilution factor (generally being 3 dilution factors reciprocal) of required mensuration drips in aseptic plate, pours the beef-protein medium that aseptic temperature is bathed into, leaves standstill behind the mixing and solidifies, and makees 6 plates altogether, and each dilution factor repeats 2 times;
4. cultivate: treat to put upside down in incubator after plate solidifies, cultivated 16 hours for 37 ℃;
5. colony counting: cultivate the back and observe the growing state of each plate bacterium colony and on plate, put the note clump count, overlap if any 2 or 2 above bacterium colonies, should all count, notice that the plate clump count should be between 20-300, otherwise should adjust diluent, redeterminate, behind the some meter, by formula calculate viable count, fill in record;
6. computing formula:
Viable count (cfu/ml)=2 plate clump count sum/2 * 5 * dilution factor
The viable count testing result of the preparation that room temperature is placed is as shown in table 1, and the result shows, according to the living bacterial liquid body preparation of the method preparation of embodiment 1, room temperature after 12 months viable count still 2 * 10
9More than the cfu/ml;
The viable count testing result of the active bacteria formulation of 37 ℃ of placements is as shown in table 2, and the result shows, uses viable bacteria goods of the present invention, places after 6 months for 37 ℃ still to be higher than 2 * 10
9More than the cfu/ml.
The active bacteria formulation room temperature preservation situation of table 1 the present invention preparation
Annotate: "-" expression viable count is lower than 1.0 * 10 in the table
9Cfu/ml.
The result shows that bacillus subtilis active bacteria formulation of the present invention is preserved 12 months viable counts at normal temperatures and still remained on 2.0 * 10
9Cfu/ml preserves 6 months viable counts and still remains on 4.9 * 10 under the environment of accelerated test temperature higher (37 ℃)
9Cfu/ml.
Claims (3)
1. a protective agent that is used to prepare the bacillus subtilis active bacteria formulation is characterized in that being made up of following component (W/V): lactose 3-4.5%, sodium carboxymethyl cellulose 0.5-1%, potassium sorbate 0.5-1%, vitamin E 0.1-0.5%, vitamin C 0.1-0.5%, water quality Fructus Citri Limoniae 0.1-0.2%.
2. protective agent according to claim 1 is characterized in that its described constituent content (W/V) is preferably lactose 4.5%, sodium carboxymethyl cellulose 0.5%, potassium sorbate 0.5%, vitamin E 0.1%, vitamin C 0.1%, water quality Fructus Citri Limoniae 0.1%.
3. the protective agent of bacillus subtilis active bacteria formulation according to claim 1 and 2 is characterized in that containing 1.0 * 10 in every 1ml protective agent
9-1.0 * 10
14Cfu.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100703097A CN102000343A (en) | 2009-09-01 | 2009-09-01 | Protective agent of live bacillus subtilis preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100703097A CN102000343A (en) | 2009-09-01 | 2009-09-01 | Protective agent of live bacillus subtilis preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102000343A true CN102000343A (en) | 2011-04-06 |
Family
ID=43808211
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100703097A Pending CN102000343A (en) | 2009-09-01 | 2009-09-01 | Protective agent of live bacillus subtilis preparation |
Country Status (1)
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| CN (1) | CN102000343A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283938A (en) * | 2012-10-23 | 2013-09-11 | 广州格拉姆生物科技有限公司 | Method for preparing bacillus subtilis active microbial agent |
| CN104212716A (en) * | 2013-05-30 | 2014-12-17 | 北京世纪阿姆斯生物技术股份有限公司 | Jelly-liked paenibacillus industrialization protective agent |
| CN113016825A (en) * | 2021-05-18 | 2021-06-25 | 郭云 | Bacillus subtilis preparation and production method thereof |
| CN113462573A (en) * | 2021-07-12 | 2021-10-01 | 河北科技大学 | Preservation method of agricultural bacillus liquid microbial inoculum |
-
2009
- 2009-09-01 CN CN2009100703097A patent/CN102000343A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103283938A (en) * | 2012-10-23 | 2013-09-11 | 广州格拉姆生物科技有限公司 | Method for preparing bacillus subtilis active microbial agent |
| CN104212716A (en) * | 2013-05-30 | 2014-12-17 | 北京世纪阿姆斯生物技术股份有限公司 | Jelly-liked paenibacillus industrialization protective agent |
| CN104212716B (en) * | 2013-05-30 | 2019-01-25 | 北京世纪阿姆斯生物技术有限公司 | A kind of gel-shaped series bacillus industrialization protective agent |
| CN113016825A (en) * | 2021-05-18 | 2021-06-25 | 郭云 | Bacillus subtilis preparation and production method thereof |
| CN113462573A (en) * | 2021-07-12 | 2021-10-01 | 河北科技大学 | Preservation method of agricultural bacillus liquid microbial inoculum |
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Application publication date: 20110406 |