CN110295113A - Marine protease bacillus stability protective agent - Google Patents
Marine protease bacillus stability protective agent Download PDFInfo
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- CN110295113A CN110295113A CN201910694814.2A CN201910694814A CN110295113A CN 110295113 A CN110295113 A CN 110295113A CN 201910694814 A CN201910694814 A CN 201910694814A CN 110295113 A CN110295113 A CN 110295113A
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- concentration
- protective agent
- tris
- mannitol
- edta
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- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 31
- 108091005804 Peptidases Proteins 0.000 title claims abstract description 31
- 239000004365 Protease Substances 0.000 title claims abstract description 31
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title claims abstract description 31
- 239000003223 protective agent Substances 0.000 title claims abstract description 20
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 62
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 42
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims abstract description 38
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 25
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 21
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 21
- 229930195725 Mannitol Natural products 0.000 claims abstract description 21
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 21
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 21
- 239000000594 mannitol Substances 0.000 claims abstract description 21
- 235000010355 mannitol Nutrition 0.000 claims abstract description 21
- 239000001103 potassium chloride Substances 0.000 claims abstract description 19
- 235000011164 potassium chloride Nutrition 0.000 claims abstract description 19
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims abstract description 14
- 239000000872 buffer Substances 0.000 claims description 26
- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 claims description 24
- 229910052787 antimony Inorganic materials 0.000 claims description 14
- 235000010290 biphenyl Nutrition 0.000 claims description 14
- 239000004305 biphenyl Substances 0.000 claims description 14
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 claims description 14
- OBETXYAYXDNJHR-SSDOTTSWSA-M (2r)-2-ethylhexanoate Chemical compound CCCC[C@@H](CC)C([O-])=O OBETXYAYXDNJHR-SSDOTTSWSA-M 0.000 claims description 13
- 239000011814 protection agent Substances 0.000 claims description 13
- 239000004615 ingredient Substances 0.000 claims description 7
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000460 chlorine Substances 0.000 claims description 2
- 229910052801 chlorine Inorganic materials 0.000 claims description 2
- 239000011591 potassium Substances 0.000 claims description 2
- 229910052700 potassium Inorganic materials 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 12
- -1 2-ethyl diphenyl antimony hexanoate Chemical compound 0.000 abstract description 8
- 239000007788 liquid Substances 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 2
- 239000007853 buffer solution Substances 0.000 abstract 2
- 241000006384 Jeotgalibacillus marinus Species 0.000 abstract 1
- 230000036425 denaturation Effects 0.000 abstract 1
- 238000004925 denaturation Methods 0.000 abstract 1
- 238000005057 refrigeration Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 28
- 241000894006 Bacteria Species 0.000 description 18
- 238000004519 manufacturing process Methods 0.000 description 9
- 229910052739 hydrogen Inorganic materials 0.000 description 7
- 239000001257 hydrogen Substances 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 6
- 239000011574 phosphorus Substances 0.000 description 6
- 229910052698 phosphorus Inorganic materials 0.000 description 6
- YFEPPSDKRDHTLB-UHFFFAOYSA-N diphenylantimony Chemical compound C=1C=CC=CC=1[Sb]C1=CC=CC=C1 YFEPPSDKRDHTLB-UHFFFAOYSA-N 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 235000014786 phosphorus Nutrition 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 4
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000040710 Chela Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- WATWJIUSRGPENY-UHFFFAOYSA-N antimony atom Chemical compound [Sb] WATWJIUSRGPENY-UHFFFAOYSA-N 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
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
- C12N1/04—Preserving or maintaining viable microorganisms
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a bacillus marinus stability protective agent, belonging to the technical field of microorganisms. The protective agent comprises Tris-HCl buffer solution, glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate and 2-ethyl diphenyl antimony hexanoate, wherein the Tris-HCl buffer solution, the glycerol, the mannitol, the EDTA, the potassium chloride, the magnesium sulfate, the disodium hydrogen phosphate and the 2-ethyl diphenyl antimony hexanoate are prepared into a liquid protective agent according to a certain proportion. The stability protective agent for the marine protease bacillus overcomes the problems of denaturation and low survival rate of marine protease bacillus strains under the condition of low-temperature refrigeration, and can stabilize the survival rate of the strains to be more than 90%.
Description
Technical field
The present invention relates to the protections that microorganism preservation field more particularly to a kind of ocean produce protease bacillus stability
Agent.
Background technique
With scale, the rapid development of intensive aquaculture industry, aquiculture disease frequently occurs.In recent years, people
Begin trying that cultivation life can be improved using the producing enzyme probiotics in beneficial microbe, especially ocean in breeding production
State environment improves the immunity of cultivated animals, inhibits pathogenic microorganism, to reduce the generation of disease.Wherein, source Yu Haiyang
The bacillus for producing protease produces the property of protease because its is distinctive, is developed diversified usage mode, Ke Yizhi
It connects and breeding water body or be added in feed is added uses, marine cultured animal can also be acted on by food organisms, it can be with
It is used by injection or dipping bath.
Due to such production protease bacillus source Yu Haiyang, growing survival condition should be with marine eco-environment phase
Seemingly, but usually it is low, active that the strain turned out under the environmental condition of laboratory simulation will appear survival rate when stored refrigerated
The problems such as decline.Therefore, seeking a kind of ocean white protective agent for not having bacillus stability of laying eggs that is able to maintain is correlative study
The project that personnel are probing into always.
Summary of the invention
The object of the present invention is to provide a kind of oceans to produce the stability protection agent of protease bacillus, is increased using the protective agent
The activity of strong bacterium and the survival rate for improving bacterium.
To realize the above-mentioned technical purpose, the technical solution adopted by the present invention are as follows: a kind of ocean production protease bacillus is steady
Qualitative protective agent, it is characterised in that: guarantor's protective agent is by adding glycerol, mannitol, EDTA, chlorine into Tris-HCl buffer
Change potassium, magnesium sulfate, disodium hydrogen phosphate and 2 ethyl hexanoic acid diphenyl antimony to be made.
Further, the Tris-HCl buffer refers to that 10 mM Tris-HCl buffers, the pH value of buffer are
7.5。
Further, the concentration of the glycerol is 25-35%(v/v), the concentration of the mannitol is 5.5-7.5%(w/v),
The concentration of the EDTA is 1.5-3.5%(w/v), the potassium chloride concentration be 0.5-0.7%(w/v), the magnesium sulfate concentration be
0.35-0.55%(w/v), the disodium hydrogen phosphate concentration is 0.25-0.45%(w/v), the 2 ethyl hexanoic acid diphenyl antimony
Concentration is 0.03-0.08%(v/v), the concentration of above-mentioned each ingredient is calculated on the basis of protectant volume.
Further, the concentration of glycerol is 32%(v/v), the concentration of the mannitol is 5.5%(w/v), the EDTA
Concentration is 1.5%(w/v), the potassium chloride concentration be 0.55%(w/v), the magnesium sulfate concentration be 0.35%(w/v), the phosphorus
Sour disodium hydrogen concentration is 0.32%(w/v), the concentration of the 2 ethyl hexanoic acid diphenyl antimony be 0.05%(v/v).
Further, the stabilizer is the preparation method is as follows: glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphoric acid hydrogen
Disodium, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5), normal
It is stirred evenly under temperature, is completely dissolved solute, after standing 10 min, with the solution of 10 mM Tris-HCl buffers (pH 7.5)
Constant volume.
A kind of ocean of the present patent application produces the stability protection agent of protease bacillus, specifically buffers in Tris-HCl
Liquid is to be added with glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate, 2 ethyl hexanoic acid diphenyl in basic liquid
Antimony.Wherein, Tris-HCl buffer makes the pH of solution keep stablizing for providing stable liquid environment, and pH7.6 close to
Ocean produces the growth pH environment of protease bacillus in the seawater；Glycerol is intracellular for being freely spread to, and reduces intracellular
Water activity, inhibit bacterial strain biochemical reaction progress；Mannitol is used to balance the osmotic pressure inside and outside strain cell；EDTA is as chela
Antibacterial agent is closed, for inhibiting varied bacteria growing；Potassium chloride, magnesium sulfate, disodium hydrogen phosphate can provide ocean and produce protease bacillus
The inorganic ion of required consumption in marine environment；2 ethyl hexanoic acid diphenyl antimony effectively can inhibit strain to fail, and be a kind of
Important biostability.The activity and survival rate of protease bacillus can be greatly improved by protectant application.
Specific embodiment
Below by specific embodiment to ocean provided by the invention produce the stability protection agent of protease bacillus do into
One step explanation.
Embodiment 1
A kind of ocean production protease bacillus stability protection agent, takes glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphorus
Sour disodium hydrogen, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5),
It stirs evenly at normal temperature, is completely dissolved solute, after standing 10 min, with 10 mM Tris-HCl buffers (pH 7.5)
Solution constant volume obtains the concentration of each ingredient in final protective agent are as follows: 32% glycerol (v/v), 5.50% mannitol (w/v), 1.50%
EDTA(w/v), 0.55% potassium chloride (w/v), 0.35% magnesium sulfate (w/v), 0.32% disodium hydrogen phosphate (w/v), 0.05% 2- ethyl
Caproic acid diphenyl antimony (v/v).Wherein mM unit meaning is mM every liter.
It takes the ocean that viable bacteria content is 5,000,000,000 cfu/mL to produce protease bacillus living bacterium solution, is added and stablizes into bacterium solution
Property protective agent 5%, saved 30 days under the conditions of 4 DEG C, every 5 days take out be coated on 2216E plate, be placed in 30 DEG C of constant temperature incubations
24 h are cultivated in case, carry out count plate, and statistics survival rate is as shown in the table.
Embodiment 2
A kind of ocean production protease bacillus stability protection agent, takes glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphorus
Sour disodium hydrogen, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5),
It stirs evenly at normal temperature, is completely dissolved solute, after standing 10 min, with 10 mM Tris-HCl buffers (pH 7.5)
Solution constant volume obtains the concentration of each ingredient in final protective agent are as follows: 35% glycerol (v/v), 6.2% mannitol (w/v), 3.20%
EDTA(w/v), 0.50% potassium chloride (w/v), 0.47% magnesium sulfate (w/v), 0.30% disodium hydrogen phosphate (w/v), 0.06% 2- ethyl
Caproic acid diphenyl antimony (v/v), with the solution constant volume of 10 mM Tris-HCl buffers (pH 7.5).Wherein mM unit meaning is milli
Mole every liter.
It takes the ocean that viable bacteria content is 5,000,000,000 cfu/mL to produce protease bacillus living bacterium solution, is added and stablizes into bacterium solution
Property protective agent 5%, saved 30 days under the conditions of 4 DEG C, every 5 days take out be coated on 2216E plate, be placed in 30 DEG C of constant temperature incubations
24 h are cultivated in case, carry out count plate, and statistics survival rate is as shown in the table.
Embodiment 3
A kind of ocean production protease bacillus stability protection agent, takes glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphorus
Sour disodium hydrogen, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5),
It stirs evenly at normal temperature, is completely dissolved solute, after standing 10 min, with 10 mM Tris-HCl buffers (pH 7.5)
Solution constant volume obtains the concentration of each ingredient in final protective agent are as follows: 27% glycerol (v/v), 5.8% mannitol (w/v), 1.8%
EDTA(w/v), 0.60% potassium chloride (w/v), 0.55% magnesium sulfate (w/v), 0.45% disodium hydrogen phosphate (w/v), 0.07% 2- ethyl
Caproic acid diphenyl antimony (v/v), with the solution constant volume of 10 mM Tris-HCl buffers (pH 7.5).Wherein mM unit meaning is milli
Mole every liter.
It takes the ocean that viable bacteria content is 5,000,000,000 cfu/mL to produce protease bacillus living bacterium solution, is added and stablizes into bacterium solution
Property protective agent 5%, saved 30 days under the conditions of 4 DEG C, every 5 days take out be coated on 2216E plate, be placed in 30 DEG C of constant temperature incubations
24 h are cultivated in case, carry out count plate, and statistics survival rate is as shown in the table.
Embodiment 4
A kind of ocean production protease bacillus stability protection agent, takes glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphorus
Sour disodium hydrogen, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5),
It stirs evenly at normal temperature, is completely dissolved solute, after standing 10 min, with 10 mM Tris-HCl buffers (pH 7.5)
Solution constant volume obtains the concentration of each ingredient in final protective agent are as follows: 30% glycerol (v/v), 6.5% mannitol (w/v), 3.4%
EDTA(w/v), 0.47% potassium chloride (w/v), 0.45% magnesium sulfate (w/v), 0.35% disodium hydrogen phosphate (w/v), 0.08% 2- ethyl
Caproic acid diphenyl antimony (v/v), with the solution constant volume of 10 mM Tris-HCl buffers (pH 7.5).Wherein mM unit meaning is milli
Mole every liter.
It takes the ocean that viable bacteria content is 5,000,000,000 cfu/mL to produce protease bacillus living bacterium solution, is added and stablizes into bacterium solution
Property protective agent 5%, saved 30 days under the conditions of 4 DEG C, every 5 days take out be coated on 2216E plate, be placed in 30 DEG C of constant temperature incubations
24 h are cultivated in case, carry out count plate, and statistics survival rate is as shown in the table.
Embodiment 5
A kind of ocean production protease bacillus stability protection agent, takes glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, phosphorus
Sour disodium hydrogen, 2 ethyl hexanoic acid diphenyl antimony are added in proportion in the solution of 10 mM Tris-HCl buffers (pH 7.5),
It stirs evenly at normal temperature, is completely dissolved solute, after standing 10 min, with 10 mM Tris-HCl buffers (pH 7.5)
Solution constant volume obtains the concentration of each ingredient in final protective agent are as follows: 25% glycerol (v/v), 5.8% mannitol (w/v), 3.5%
EDTA(w/v), 0.70% potassium chloride (w/v), 0.50% magnesium sulfate (w/v), 0.35% disodium hydrogen phosphate (w/v), 0.04% 2- ethyl
Caproic acid diphenyl antimony (v/v), with the solution constant volume of 10 mM Tris-HCl buffers (pH 7.5).Wherein mM unit meaning is milli
Mole every liter.
It takes the ocean that viable bacteria content is 5,000,000,000 cfu/mL to produce protease bacillus living bacterium solution, is added and stablizes into bacterium solution
Property protective agent 5%, saved 30 days under the conditions of 4 DEG C, every 5 days take out be coated on 2216E plate, be placed in 30 DEG C of constant temperature incubations
24 h are cultivated in case, carry out count plate, and statistics survival rate is as shown in the table.
Inventor is by the analysis of comparison above embodiments as a result, finding that ocean of the invention produces protease bacillus stability and protects
Shield agent can keep ocean to produce protease bacillus survival rate and be maintained at 90% or more, wherein the effect of embodiment 1 is best, reaches
To 94.16%.
Claims (4)
1. a kind of ocean produces the stability protection agent of protease bacillus, it is characterised in that: guarantor's protective agent is by Tris-
Glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate and 2 ethyl hexanoic acid diphenyl antimony are added in HCl buffer
It is made.
2. a kind of ocean according to claim 1 produces the stability protection agent of protease bacillus, it is characterised in that: described
Tris-HCl buffer refers to that 10 mM Tris-HCl buffers, the pH value of buffer are 7.5.
3. a kind of ocean according to claim 2 produces the stability protection agent of protease bacillus, it is characterised in that: described
The concentration of glycerol is 25-35%(v/v), the concentration of the mannitol is 5.5-7.5%(w/v), the concentration of the EDTA be 1.5-
3.5%(w/v), the potassium chloride concentration is 0.5-0.7%(w/v), the magnesium sulfate concentration be 0.35-0.55%(w/v), it is described
Disodium hydrogen phosphate concentration is 0.25-0.45%(w/v), the concentration of the 2 ethyl hexanoic acid diphenyl antimony be 0.03-0.08%(v/
V), the concentration of above-mentioned each ingredient is calculated on the basis of protectant volume.
4. a kind of ocean according to claim 3 produces the stability protection agent of protease bacillus, it is characterised in that: glycerol
Concentration be 32%(v/v), the concentration of the mannitol is 5.5%(w/v), the concentration of the EDTA be 1.5%(w/v), the chlorine
Change potassium concn is 0.55%(w/v), the magnesium sulfate concentration be 0.35%(w/v), the disodium hydrogen phosphate concentration be 0.32%(w/
V), the concentration of the 2 ethyl hexanoic acid diphenyl antimony is 0.05%(v/v).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910694814.2A CN110295113B (en) | 2019-07-30 | 2019-07-30 | Marine protease bacillus stability protective agent |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910694814.2A CN110295113B (en) | 2019-07-30 | 2019-07-30 | Marine protease bacillus stability protective agent |
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| Publication Number | Publication Date |
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| CN110295113A true CN110295113A (en) | 2019-10-01 |
| CN110295113B CN110295113B (en) | 2022-05-13 |
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ID=68032189
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA870647A (en) * | 1971-05-11 | R. Leebrick John | Diphenylantimony 2-ethylhexanoate and preparation | |
| WO2000007606A2 (en) * | 1998-08-07 | 2000-02-17 | Ganeden Biotech, Inc. | Methods for increasing the solubility of nutritional materials using probiotic lactic acid-producing bacteria |
| JP2003320000A (en) * | 2002-05-02 | 2003-11-11 | Fuji Silysia Chemical Ltd | Sterilizing method and sterilizing apparatus |
| WO2004063036A1 (en) * | 2003-01-09 | 2004-07-29 | Baxter Healthcare S.A. | Safety containers for biologically active substances and method for producing said container |
| CN101511950A (en) * | 2006-08-29 | 2009-08-19 | 3M创新有限公司 | Resin systems including reactive surface-modified nanoparticles |
-
2019
- 2019-07-30 CN CN201910694814.2A patent/CN110295113B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA870647A (en) * | 1971-05-11 | R. Leebrick John | Diphenylantimony 2-ethylhexanoate and preparation | |
| WO2000007606A2 (en) * | 1998-08-07 | 2000-02-17 | Ganeden Biotech, Inc. | Methods for increasing the solubility of nutritional materials using probiotic lactic acid-producing bacteria |
| JP2003320000A (en) * | 2002-05-02 | 2003-11-11 | Fuji Silysia Chemical Ltd | Sterilizing method and sterilizing apparatus |
| WO2004063036A1 (en) * | 2003-01-09 | 2004-07-29 | Baxter Healthcare S.A. | Safety containers for biologically active substances and method for producing said container |
| CN101511950A (en) * | 2006-08-29 | 2009-08-19 | 3M创新有限公司 | Resin systems including reactive surface-modified nanoparticles |
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| Publication number | Publication date |
|---|---|
| CN110295113B (en) | 2022-05-13 |
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