CN102181389B - Method for breeding Bacillaceae with high spore yield - Google Patents
Method for breeding Bacillaceae with high spore yield Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 20
- 241001112741 Bacillaceae Species 0.000 title claims abstract description 19
- 238000009395 breeding Methods 0.000 title claims abstract description 12
- 230000001488 breeding effect Effects 0.000 title claims abstract description 12
- 238000010438 heat treatment Methods 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims description 75
- 230000001954 sterilising effect Effects 0.000 claims description 62
- 238000004659 sterilization and disinfection Methods 0.000 claims description 58
- 241000726221 Gemma Species 0.000 claims description 37
- 230000003068 static effect Effects 0.000 claims description 37
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 29
- 241000194108 Bacillus licheniformis Species 0.000 claims description 7
- 241000194107 Bacillus megaterium Species 0.000 claims description 7
- 244000063299 Bacillus subtilis Species 0.000 claims description 6
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 6
- 241000193755 Bacillus cereus Species 0.000 claims description 5
- 241000193749 Bacillus coagulans Species 0.000 claims description 5
- 241000193403 Clostridium Species 0.000 claims description 5
- 229940054340 bacillus coagulans Drugs 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
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- 230000004913 activation Effects 0.000 claims 1
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- 230000004151 fermentation Effects 0.000 abstract description 6
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- 230000003213 activating effect Effects 0.000 abstract 1
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- 230000002035 prolonged effect Effects 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 50
- 238000011081 inoculation Methods 0.000 description 32
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- 239000002504 physiological saline solution Substances 0.000 description 24
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- 239000007788 liquid Substances 0.000 description 19
- 235000015097 nutrients Nutrition 0.000 description 19
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 14
- 235000011187 glycerol Nutrition 0.000 description 7
- 238000010186 staining Methods 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241001478240 Coccus Species 0.000 description 2
- 241000186541 Desulfotomaculum Species 0.000 description 2
- 241000605909 Fusobacterium Species 0.000 description 2
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- 241001465754 Metazoa Species 0.000 description 2
- 241000192023 Sarcina Species 0.000 description 2
- 241000204117 Sporolactobacillus Species 0.000 description 2
- WJJMNDUMQPNECX-UHFFFAOYSA-N dipicolinic acid Chemical compound OC(=O)C1=CC=CC(C(O)=O)=N1 WJJMNDUMQPNECX-UHFFFAOYSA-N 0.000 description 2
- 241000894007 species Species 0.000 description 2
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- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for breeding Bacillaceae with high spore yield. In the method, the Bacillaceae with high spore yield is bred in a way of performing heating treatment stage by stage and then culturing according to the thermal stability characteristic of spores. The method comprises the following steps of: activating Bacillaceae, performing first-stage heating treatment, performing first-stage culture, performing second-stage heating treatment, performing second-stage culture, performing third-stage heating treatment, performing third-stage culture, and collecting, wherein in the first-stage heating treatment, heating conditions are that the Bacillaceae is heated for 1 to 6h in water bath at the temperature of between 50 and 60 DEG C; in the second-stage heating treatment, heating conditions are that the Bacillaceae is heated for 10 to 60min in water bath at the temperature of between 70 and 80 DEG C; and in the third-stage heating treatment, heating conditions are that the Bacillaceae is heated for 1 to 5min in water bath at the temperature of between 90 and 100 DEG C. The Bacillaceae can quickly form the spores in the fermentation production process, so that the fermentation time is shortened, the production cost is reduced, the preservation time of finished products is prolonged, and the using effect is improved. Moreover, the process is simple, the cost is low, and the method has the advantage of strong operability.
Description
Technical field
The present invention relates to the method for a kind of seed selection genus bacillus, specifically a kind of method of breeding Bacillaceae with high spore yield belongs to the Biotechnology and Bioengineering field.
Background technology
Genus bacillus (Bacillaceae), the section of bacterium can form bacillus or the coccus of gemma (statospore).Be generally aerobic or amphimicrobian, gramstaining is positive.Comprise bacillus, Sporolactobacillus, fusobacterium, Desulfotomaculum and gemma Sarcina etc.Under certain environment, endobacillary structure changes, and through the prespore stage, forms a complete gemma.Gemma has very strong resistibility to heat, radioactive rays and chemical substance etc.Aspect chemical constitution, contain the calcium salt of non-existent dipicolinic acid in a large amount of vegetative cells at gemma; In configuration aspects, the protoplasma of gemma is with trilamellar membrane outward, is thick cortex, spore coat and exine from inside to outside.In bacillus, to Species Division be the shape of size, spore with thalline and in endobacillary position, utilization and the product thereof of sugar, can go back orthonitric acid, and can grow under the salt condition of high density that to wait be foundation.It extensively is distributed in water, air and the soil and animal intestinal etc. is located.Representative species is subtilis, bacillus natto, Bacillus licheniformis, cured shape genus bacillus, bacillus megaterium etc.Genus bacillus is widely used, and wherein subtilis, Bacillus licheniformis can be used for animal husbandry and fishery fodder additives and water surrounding purification; The Su Yun genus bacillus can be used for plant protection, kill pests; Bacillus megaterium has the solid potassium effect of phosphorus decomposing and is used for producing biological organic fertilizer; Bacillus polymyxa has the ability of fixed member attitude nitrogen.
Whether genus bacillus can form the effect of stablizing the whole production of gemma state relation and later stage preservation use thereof by rapid, high volume in processing, wherein the genus bacillus kind of high yield gemma rate seems and is even more important, and overlaps fast and effectively that the method for breeding Bacillaceae with high spore yield is very necessary so take measures to set up one.
Summary of the invention
The object of the present invention is to provide a kind of method of breeding Bacillaceae with high spore yield.
Technical scheme of the present invention: according to the thermostability characteristics of gemma, adopt stepped heating to process rear means of cultivating, select the genus bacillus of high yield gemma rate, thereby can in fermentation production process, shorten fermentation time, reduce production costs, prolong the finished product shelf time, improve result of use.Realize the technical program, may further comprise the steps:
1. genus bacillus is activated: will treat the aseptic streak inoculation of genus bacillus of seed selection on the solid medium of sterilization, 30～38 ℃ of static cultivations 3～5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place water-bath water-bath 1～6h of 50～60 ℃;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30～38 ℃ of static cultivations 3～5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place water-bath water-bath 10～60min of 70～80 ℃;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30～38 ℃ of static cultivations 3～5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place water-bath water-bath 1～5min of 90～100 ℃;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, and the bacterium colony on the solid medium is collected in 30～38 ℃ of static cultivations 5～7 days, obtains high yield gemma rate genus bacillus kind;
The solid medium of described sterilization is the substratum of same prescription, and its component comprises by mass percentage: extractum carnis 0.30%, sodium-chlor 0.50%, peptone 1.00%, agar 2.00%, and pH 7.2; Its sterilising conditions is 121 ℃, 0.11Mpa, sterilization 20～30min;
Described genus bacillus is for forming bacillus or the coccus of gemma (statospore), comprise bacillus, Sporolactobacillus, fusobacterium, Desulfotomaculum and gemma Sarcina etc., preferred preserving number is the subtilis of ATCC6633, preserving number is the Bacillus licheniformis of ATCC 11946, preserving number is the bacillus megaterium of ATCC14581, preserving number is the Bacillus coagulans of ATCC 7050, preserving number is the bacillus cereus of ATCC14579, preserving number is that bacillus natto and the preserving number of AS 1.1086 is the clostridium butylicum of ATCC 8260.
The method of a kind of breeding Bacillaceae with high spore yield of the present invention has following beneficial effect:
Use the present invention to select the genus bacillus of high yield gemma rate, can in fermentation production process, form fast gemma, thereby shorten fermentation time, reduce production costs, prolong the finished product shelf time, improve result of use.And technique is simple, with low cost, has advantages of workable.
Embodiment:
In order better to understand the present invention, below the invention will be further described with embodiment.
Embodiment 1:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of subtilis in the glycerine (ATCC 6633) on the solid medium of sterilization, 30 ℃ of static cultivations 4 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 55 ℃ water-bath water-bath 3h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 4 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 75 ℃ water-bath water-bath 30min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 4 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 95 ℃ water-bath water-bath 2min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 30 ℃ of static cultivations obtain the subtilis (solid medium of described sterilization of high gemma rate after 5 days, its component comprises by mass percentage: extractum carnis 0.30%, sodium-chlor 0.50%, peptone 1.00%, agar 2.00%, and pH 7.2; Its sterilising conditions is 121,0.11Mpa, sterilization 25min);
The high gemma rate subtilis that obtains is inoculated in (the liquid nutrient medium of described sterilization on the liquid nutrient medium of sterilization, its component comprises by mass percentage: extractum carnis 0.30%, peptone 1.00%, sodium-chlor 0.50%, yeast extract paste 0.04%, iron(ic) chloride 0.02%, calcium carbonate 0.02%, manganous sulfate 0.01%, and pH 7.2; Its sterilising conditions is 121 ℃, 0.11Mpa, sterilization 25min), after 30 ℃, 150rpm are cultivated 48h, violet staining oil sem observation, 96% forms gemma.
Embodiment 2:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of Bacillus licheniformis in the glycerine (ATCC 11946) on the solid medium of sterilization, 37 ℃ of static cultivations 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 60 ℃ water-bath water-bath 5h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 37 ℃ of static cultivations 5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 78 ℃ water-bath water-bath 35min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 37 ℃ of static cultivations 5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 96 ℃ water-bath water-bath 2min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 37 ℃ of static cultivations are after 7 days, obtain the Bacillus licheniformis (prescription of the solid medium of described sterilization is with the solid medium of embodiment 1) of high gemma rate;
The Bacillus licheniformis of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with embodiment 1 liquid nutrient medium), after 37 ℃, 150rpm are cultivated 72h, violet staining oil sem observation, 94% forms gemma.
Embodiment 3:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of bacillus megaterium in the glycerine (ATCC 14581) on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 58 ℃ water-bath water-bath 5h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 74 ℃ water-bath water-bath 40min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 98 ℃ water-bath water-bath 3min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 30 ℃ of static cultivations are after 7 days, obtain the bacillus megaterium (prescription of the solid medium of described sterilization is with the solid medium of embodiment 1) of high gemma rate;
The bacillus megaterium of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with the liquid nutrient medium of embodiment 1), after 30 ℃, 150rpm are cultivated 48h, violet staining oil sem observation, 98% forms gemma.
Embodiment 4:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of Bacillus coagulans in the glycerine (ATCC 7050) on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 60 ℃ water-bath water-bath 5h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 78 ℃ water-bath water-bath 50min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 98 ℃ water-bath water-bath 4min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 30 ℃ of static cultivations obtain the Bacillus coagulans (prescription of the solid medium of described sterilization is with embodiment 1 solid medium) of high gemma rate after 7 days;
The Bacillus coagulans of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with embodiment 1 liquid nutrient medium), after 30 ℃, 150rpm are cultivated 48h, violet staining oil sem observation, 97% forms gemma.
Embodiment 5:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of bacillus cereus in the glycerine (ATCC 14579) on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 58 ℃ water-bath water-bath 6h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 4 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 75 ℃ water-bath water-bath 50min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 4 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 97 ℃ water-bath water-bath 4min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 30 ℃ of static cultivations obtain the bacillus cereus (prescription of the solid medium of described sterilization is with the solid medium of embodiment 1) of high gemma rate after 6 days;
The bacillus cereus of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with embodiment 1 solid medium), after 30 ℃, 150rpm are cultivated 48h, violet staining oil sem observation, 96% forms gemma.
Embodiment 6:
1. genus bacillus is activated: will-70 ℃ be stored in the aseptic streak inoculation of bacillus natto in the glycerine (AS 1.1086) on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 56 ℃ water-bath water-bath 6h;
3. one-level is cultivated: the aseptic streak inoculation of bacteria suspension in will be 2. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 80 ℃ water-bath water-bath 50min;
5. secondary is cultivated: the aseptic streak inoculation of bacteria suspension in will be 4. in the test tube on the solid medium of sterilization, 30 ℃ of static cultivations 5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 98 ℃ water-bath water-bath 3min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 30 ℃ of static cultivations obtain the bacillus natto (prescription of the solid medium of described sterilization is with embodiment 1 solid medium) of high gemma rate after 7 days;
The bacillus natto of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with embodiment 1 liquid nutrient medium), after 30 ℃, 150rpm are cultivated 48h, violet staining oil sem observation, 95% forms gemma.
Embodiment 7:
1. genus bacillus is activated: incite somebody to action-70 ℃ and be stored in the aseptic streak inoculation of clostridium butylicum in the glycerine (ATCC 8260) on the solid medium of sterilization, 37 ℃ of static anaerobism were cultivated 5 days;
2. one-level heat treated: be contained in after the bacterium colony in will be 1. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 55 ℃ water-bath water-bath 5h;
3. one-level is cultivated: 2. the aseptic streak inoculation of bacteria suspension in the middle test tube is on the solid medium of sterilization, and 37 ℃ of static anaerobism were cultivated 5 days;
4. secondary heat treated: be contained in after the bacterium colony in will be 3. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 75 ℃ water-bath water-bath 50min;
5. secondary is cultivated: 4. the aseptic streak inoculation of bacteria suspension in the middle test tube is on the solid medium of sterilization, and 37 ℃ of static anaerobism were cultivated 5 days;
6. three grades of heat treated: be contained in after the bacterium colony in will be 5. on the solid medium washes with stroke-physiological saline solution in the sterile test tube with silica gel plug, place 98 ℃ water-bath water-bath 4min;
7. three grades of cultivations and collection: the aseptic streak inoculation of bacteria suspension in will be 6. in the test tube is on the solid medium of sterilization, 37 ℃ of static anaerobism are cultivated the clostridium butylicum (prescription of the solid medium of described sterilization is with embodiment 1 solid medium) that obtains high gemma rate after 7 days;
The clostridium butylicum of this high gemma rate is inoculated on the liquid nutrient medium of sterilization (prescription of the liquid nutrient medium of described sterilization is with embodiment 1 liquid nutrient medium), after 37 ℃, static anaerobism are cultivated 48h, violet staining oil sem observation, 98% forms gemma.
Claims (5)
1. the method for a breeding Bacillaceae with high spore yield, the means of cultivating after it is characterized in that adopting stepped heating to process, select the genus bacillus of high yield gemma rate, its step comprises: genus bacillus activation, one-level heat treated, one-level cultivation, secondary heat treated, secondary cultivate, three grades of heat treated, three grades of cultivations and collection;
Described one-level heat treated, its heating condition are heating in water bath 1～6h of 50～60 ℃;
Described secondary heat treated, its heating condition are heating in water bath 10～60min of 70～80 ℃;
Described three grades of heat treated, its heating condition are heating in water bath 1～5min of 90～100 ℃;
Described genus bacillus is: preserving number is that the subtilis of ATCC 6633, the Bacillus licheniformis that preserving number is ATCC11946, the bacillus megaterium that preserving number is ATCC 14581, the Bacillus coagulans that preserving number is ATCC 7050, the bacillus cereus that preserving number is ATCC 14579, bacillus natto or the preserving number that preserving number is AS 1.1086 are the clostridium butylicum of ATCC 8260.
2. the method for a kind of breeding Bacillaceae with high spore yield according to claim 1 is characterized in that the culture condition that described one-level is cultivated is 30～38 ℃ of static cultivations 3～5 days; The culture condition that described secondary is cultivated is 30～38 ℃ of static cultivations 3～5 days; The culture condition of described three grades of cultivations is 30～38 ℃ of static cultivations 5～7 days.
3. the method for a kind of breeding Bacillaceae with high spore yield according to claim 1 is characterized in that the culture condition that described genus bacillus is activated is 30～38 ℃ of static cultivations 3～5 days.
4. the method for the described a kind of breeding Bacillaceae with high spore yield of arbitrary claim according to claim 1～3, it is characterized in that one-level is cultivated, secondary is cultivated and three grades of cultivations in used solid medium be the substratum of same prescription, this substratum contains following component by weight percentage: extractum carnis 0.30%, sodium-chlor 0.50%, peptone 1.00%, agar 2.00%, pH 7.2.
5. the method for a kind of breeding Bacillaceae with high spore yield according to claim 4, the sterilising conditions that it is characterized in that described solid medium is 121 ℃, 0.11Mpa, sterilization 20～30min.
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| CN102352329B (en) * | 2011-09-28 | 2014-03-26 | 重庆泰平药业有限公司 | Production method for microbial germ powder |
| CN102337238B (en) * | 2011-09-28 | 2014-03-26 | 重庆泰平药业有限公司 | Method for culturing microbes through fermentation |
| CN103289916A (en) * | 2013-03-20 | 2013-09-11 | 广州格拉姆生物科技有限公司 | A solid fermentation production method for Bacillus Subtilis natto |
| CN105368746B (en) * | 2015-12-01 | 2019-04-12 | 山东新华医疗器械股份有限公司 | The preparation method of bacillus stearothermophilus gemma |
| CN112941005A (en) * | 2021-01-22 | 2021-06-11 | 武汉微康益生菌研究院有限公司 | Method for rapidly producing spores by bacillus coagulans |
| CN113215056A (en) * | 2021-06-02 | 2021-08-06 | 亚太星原农牧科技海安有限公司 | Screening method of bacillus coagulans capable of easily forming spores |
| CN113564223A (en) * | 2021-07-26 | 2021-10-29 | 福建亲亲股份有限公司 | Simple identification method for bacillus cereus for konjac glucomannan |
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