CN110499268A - White staphylococcus powder and its application - Google Patents
White staphylococcus powder and its application Download PDFInfo
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- CN110499268A CN110499268A CN201910761869.0A CN201910761869A CN110499268A CN 110499268 A CN110499268 A CN 110499268A CN 201910761869 A CN201910761869 A CN 201910761869A CN 110499268 A CN110499268 A CN 110499268A
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- 239000000843 powder Substances 0.000 title claims abstract description 31
- 241000191940 Staphylococcus Species 0.000 title claims abstract description 15
- 230000001954 sterilising effect Effects 0.000 claims abstract description 70
- 238000002360 preparation method Methods 0.000 claims abstract description 37
- 239000001963 growth medium Substances 0.000 claims abstract description 31
- 238000004321 preservation Methods 0.000 claims abstract description 16
- 238000000855 fermentation Methods 0.000 claims abstract description 12
- 230000004151 fermentation Effects 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 10
- 238000002156 mixing Methods 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 9
- 230000002779 inactivation Effects 0.000 claims abstract description 8
- 238000004806 packaging method and process Methods 0.000 claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 41
- 239000006052 feed supplement Substances 0.000 claims description 39
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 34
- 240000008042 Zea mays Species 0.000 claims description 33
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 33
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 33
- 235000005822 corn Nutrition 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 31
- 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 30
- 239000008103 glucose Substances 0.000 claims description 29
- 241000894006 Bacteria Species 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 20
- 238000004090 dissolution Methods 0.000 claims description 19
- 239000001888 Peptone Substances 0.000 claims description 18
- 108010080698 Peptones Proteins 0.000 claims description 18
- 238000001514 detection method Methods 0.000 claims description 18
- 235000019319 peptone Nutrition 0.000 claims description 18
- 238000004659 sterilization and disinfection Methods 0.000 claims description 18
- 239000011780 sodium chloride Substances 0.000 claims description 17
- 238000003756 stirring Methods 0.000 claims description 17
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 15
- 239000011229 interlayer Substances 0.000 claims description 15
- 239000000284 extract Substances 0.000 claims description 13
- 239000002609 medium Substances 0.000 claims description 13
- 238000012360 testing method Methods 0.000 claims description 13
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 12
- 235000013372 meat Nutrition 0.000 claims description 12
- 238000010899 nucleation Methods 0.000 claims description 12
- 230000001079 digestive effect Effects 0.000 claims description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- 230000012010 growth Effects 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000012856 packing Methods 0.000 claims description 9
- 230000003519 ventilatory effect Effects 0.000 claims description 9
- 238000010792 warming Methods 0.000 claims description 9
- 238000009395 breeding Methods 0.000 claims description 8
- 230000001488 breeding effect Effects 0.000 claims description 8
- 238000003860 storage Methods 0.000 claims description 8
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 239000013530 defoamer Substances 0.000 claims description 7
- 229910052710 silicon Inorganic materials 0.000 claims description 7
- 239000010703 silicon Substances 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 claims description 6
- 241000233866 Fungi Species 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 238000004925 denaturation Methods 0.000 claims description 6
- 230000036425 denaturation Effects 0.000 claims description 6
- 239000006260 foam Substances 0.000 claims description 6
- 238000011534 incubation Methods 0.000 claims description 6
- 239000006916 nutrient agar Substances 0.000 claims description 6
- 235000015097 nutrients Nutrition 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 241001478240 Coccus Species 0.000 claims description 4
- 241000196324 Embryophyta Species 0.000 claims description 3
- 241000191963 Staphylococcus epidermidis Species 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 230000032683 aging Effects 0.000 claims description 3
- 230000003698 anagen phase Effects 0.000 claims description 3
- 230000000954 anitussive effect Effects 0.000 claims description 3
- 239000002518 antifoaming agent Substances 0.000 claims description 3
- 238000003556 assay Methods 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 claims description 3
- 229960001231 choline Drugs 0.000 claims description 3
- 230000007850 degeneration Effects 0.000 claims description 3
- 230000029087 digestion Effects 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 230000036512 infertility Effects 0.000 claims description 3
- 230000001788 irregular Effects 0.000 claims description 3
- 238000009533 lab test Methods 0.000 claims description 3
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- 238000011169 microbiological contamination Methods 0.000 claims description 3
- 238000000386 microscopy Methods 0.000 claims description 3
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- 229920002401 polyacrylamide Polymers 0.000 claims description 3
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- 238000011160 research Methods 0.000 claims description 3
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- 238000009777 vacuum freeze-drying Methods 0.000 claims description 3
- 230000003442 weekly effect Effects 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 229940124584 antitussives Drugs 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- PCHPORCSPXIHLZ-UHFFFAOYSA-N diphenhydramine hydrochloride Chemical compound [Cl-].C=1C=CC=CC=1C(OCC[NH+](C)C)C1=CC=CC=C1 PCHPORCSPXIHLZ-UHFFFAOYSA-N 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 claims 1
- 238000005057 refrigeration Methods 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 229940032021 tetramune Drugs 0.000 claims 1
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- 230000000694 effects Effects 0.000 abstract description 3
- 239000004615 ingredient Substances 0.000 abstract description 3
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- 235000013343 vitamin Nutrition 0.000 abstract description 3
- 229930003231 vitamin Natural products 0.000 abstract description 3
- 239000011782 vitamin Substances 0.000 abstract description 3
- 229940088594 vitamin Drugs 0.000 abstract description 3
- 150000003722 vitamin derivatives Chemical class 0.000 abstract description 3
- 239000002244 precipitate Substances 0.000 abstract 1
- 235000015099 wheat brans Nutrition 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 206010011224 Cough Diseases 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- OROGSEYTTFOCAN-DNJOTXNNSA-N codeine Chemical compound C([C@H]1[C@H](N(CC[C@@]112)C)C3)=C[C@H](O)[C@@H]1OC1=C2C3=CC=C1OC OROGSEYTTFOCAN-DNJOTXNNSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010010256 Dietary Proteins Proteins 0.000 description 1
- 102000015781 Dietary Proteins Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 230000003906 autonomic nervous system functioning Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229960004126 codeine Drugs 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 235000021245 dietary protein Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- OROGSEYTTFOCAN-UHFFFAOYSA-N hydrocodone Natural products C1C(N(CCC234)C)C2C=CC(O)C3OC2=C4C1=CC=C2OC OROGSEYTTFOCAN-UHFFFAOYSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 210000001767 medulla oblongata Anatomy 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/14—Antitussive agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Pulmonology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses White staphylococcus powder and its application comprising following steps: the preparation of step 1) strain；The preservation of step 2 strain；The preparation of step 3) culture medium；Step 4) sterilizing；Step 5) culture；Step 6) inactivation；The configuration of step 7) flocculant；Step 8) precipitates drying；Step 9) crushes；Step 10) mixing；Step 11) packaging.Culture effect of the present invention is better than conventional culture medium, and low in cost, saves cost of material to pharmacy corporation, improves the added value of industry.Vitamin and other active materials will not be destroyed, nutritional ingredient balanced and reasonable is low in cost, market can be replaced completely often to use culture medium, fermentation efficiency also correspondinglys increase.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to White staphylococcus powder and its application.
Background technique
White staphylococcus powder is a kind of oral bacterial preparation of non-pathogenic, is to be trained by non-pathogenic White staphylococcus liquid
Dry thallus is inactivated obtained by supporting to be made.Experiment shows that it has the antibechic intensity of approximate codeine, directly acts on medulla oblongata cough
It coughs maincenter, obvious to acute cough's antitussive effect, dosage is bigger, and antitussive effect is stronger, and has adjusting autonomic nervous system function,
Amount of expectoration can be made to significantly reduce.Fermentation method prepares unique way that White staphylococcus powder is pharmacy corporation production White staphylococcus powder
Diameter, but the cost of culture medium restricts the development of enterprise uses dextrose peptone medium compared with multiple enterprises at present, raw material at
This height, cell density, the profit of enterprise substantially reduces." with the optimised process item of solution fermentation production Vaccinum Bronehitidis piece
Part research, Shandong medical industry 1999 ", discloses a kind of White staphylococcus culture medium, including peptone 0.5%, urea
2%, corn pulp 1%, glucose 1%, sodium citrate 0.3%, magnesium sulfate 0.05% and sodium chloride 0.5%, the culture medium at
This is higher, and the cell concentration obtained only reaches 109CFU/mL rank.Develop a kind of White staphylococcus powder preparation of low cost
Technique is our technical issues that need to address.
Wheat bran (wheat bran) is the principal by product of flour processing, and in the past, wheat bran is primarily used to animal feed, economical
Value is not high, also has many producers arbitrarily to abandon wheat bran.Wheat bran carbohydrate rich in, protein,
(functional component of wheat bran and its application study progress, Jia Tianguang, the food such as vitamin E, phenolic compound and inorganic mineral
Research and development 2014), be gradually applied to other field, including health care product, food additives, extract dietary fiber and
Dietary proteins etc., have broad application prospect and Development volue.But due to wheat bran complicated component, treatment process is difficult to
It grasps.In Bacteria Culture field, existing research mostly uses greatly wheat bran to carry out simple process, the carbon source as bacteria culture media
Part complements, for reducing the cost of culture medium；There are no occur using wheat bran as the white grape of primary raw material so far
Coccus culture medium.
Summary of the invention
Present invention aim to address the prior art technical problem high there are fermentation costs, provide White staphylococcus powder and
It is applied, with overcome the deficiencies in the prior art.
The present invention is achieved by the following technical solutions:
White staphylococcus powder of the present invention comprising following steps:
The preparation of step 1) strain:
(1) strain: staphylococcus albus, strain are upper white 71-1, and the long white 75-1 or white 72-1 in Hunan is 1-2 plants optional, by force
Chinese institute of biological products provides；
Bacterium colony: be creamy white colony on nutrient agar, and neat in edge is opaque, not chromogenesis；
Bacterial type: should be Gram-positive, in the coccus of botryoidalis irregular alignment；
(2) fungi preservation:
| Store method | Storage temperature | Holding time |
| Vacuum freezedrying preservation method | 0-4℃ | 3-5 |
| Atoleine preservation method | 0-4℃ | 6 months |
| Periodically transplanting preservation method | 0-4℃ | 1 month |
(3) breeding of strain:
Strain in use, when there is aging, degeneration and variation phenomenon, using Pure strain separation method in time to strain into
Row breeding, rejuvenation select more satisfactory strain for producing and using；
(4) preparation of culture medium:
The preparation of nutrient agar:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml, agar 15-20g；
It takes peptone and sodium chloride to be added in meat extract, after tepor dissolution, 15-20g agar is added and boils, it is molten that glucose is added
Solution, adjusting pH value make after sterilizing to be 7.2 ± 0.2, and packing, 121 DEG C sterilize 20 minutes, cold rear bevel；
The preparation of nutrient broth medium:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml；
Peptone, sodium chloride and glucose is taken to be added in meat extract, tepor dissolution, adjusting pH value makes after sterilizing to be 7.2 ± 0.2,
Packing, 121 DEG C sterilize 20 minutes, spare after cold；
(5) preparation of strain:
By sterility requirements, the strain in freeze-drying pipe is reached several test tube slants, 37 ± 1 DEG C culture 18-24 hours, it is cold
Hiding saves, and the strain of test tube slant is passed several flat bottle inclined-planes, 37 ± 1 DEG C culture 8-10 hours, use nutrient broth medium
Or uniform bacteria suspension is made for producing and using in physiological saline, can pass several test tube slants while passing flat bottle inclined-plane, cultivate
After save, and fill in strain passage record；
The digestion of step 2) basal liquid:
(1) dosage of basal liquid: corn pulp 450kg and water 900kg (or corn water 1600kg)；
(2) basal liquid digests: corn pulp and water (or corn water) being squeezed into digester with dnockout pumps, interlayer is warming up to
At 50-60 DEG C, with NaOH tune pH value to 8.0-8.5, steaming is continuously heating to 100 DEG C in tank, keeps the temperature 1 hour, at least places 10
Hour, cold water is cooled to 50 DEG C hereinafter, with hydrochloric acid tune pH value to 6.0-6.5 before filtering, is filtered with filter press spare；
Step 3) culture medium is prepared:
(1) in feed supplement bottle each composition of feed supplement liquid proportion, dosage and preparation:
It after above each composition precise, sufficiently dissolves, is poured into feed supplement bottle respectively, added with water to full dose, after wrapping
Sterilizing；
(2) in feed supplement tank each composition of feed supplement liquid proportion, dosage and preparation
After above each composition precise, respectively change in sugar bowl, choline tank and mixer after completely dissolution, with water mend to
Required volume is pumped into respectively in each feed supplement tank with vacuum pump, carries out real tank sterilizing, can also according to actual amount, according to the proportion into
Row is prepared；
(3) culture medium is prepared:
(3.1) proportion, dosage and preparation of seed tank culture base:
| Title | Corn pulp (corn water) digestive juice | Peptone | Sodium chloride | Oral glucose | Organic silicon defoamer | Total amount |
| Proportion | 15% (or 50%) | 1% | 0.2% | 0.3% | —— | —— |
| Dosage | 90kg (or 300kg) | 6.0kg | 1.2kg | 1.8kg | 200ml | 600000 ml |
Corn pulp (or corn water) digestive juice is squeezed into seeding tank, each material is added, is mended with water to full dose, stirring is abundant
After dissolution, after sodium hydroxide tune pH value to 7.4-7.6, sterilizing；
(3.2) proportion of fermentation tank culture medium, dosage and preparation:
| Title | Corn pulp (corn water) digestive juice | Peptone | Sodium chloride | Oral glucose | Organic silicon defoamer | Total amount |
| Proportion | >=15% (or 31.4%) | 1% | 0.2% | 0.3% | —— | —— |
| Dosage | >=525kg (or 1100kg) | 35.0kg | 7.0kg | 10.5kg | 1000ml | 3500000 ml |
Corn pulp (or corn water) digestive juice is squeezed into material-compound tank, each material is added, is mended with water to prescribed volume, stirring
After completely dissolution, with after sodium hydroxide tune pH value to 7.4-7.6, the culture medium after preparation is squeezed into fermentor with dnockout pumps,
Sterilizing；Step 4) sterilizing:
(1) in feed supplement bottle feed supplement liquid and required article sterilizing:
After feed supplement bottle and extraneous connector gauze, moistureproof paper wrapping, it is put into compact hand operated sterilizer, spare
Seed bottle, after racemosus pipeline is wrapped up with same method, while sterilizing；Sterilization pressure is 0.11-0.12MPa, temperature 121-
125 DEG C, 45 minutes 40% Glucose Liquids of heat preservation will individually sterilize, sterilization pressure 0.09-0.11MPa, temperature 110-121
DEG C, keep the temperature 30 minutes；
(2) slack tank sterilizes:
Just using tank or occurs microbiological contamination with tank, interruption and need to carry out slack tank sterilization treatment；Pre-temperature tank body, interlayer are warming up to first
It 90 DEG C, opens in tank with tank bottom inlet valve, allows steam to circulate in tank after twenty minutes, turn down exhaust steam valve, rise to 0.11- to pressure
0.12MPa, temperature are 121-125 DEG C, keep the temperature 45 minutes, after sterilizing, open exhaust steam valve, are reduced to zero to pressure inside the tank
When, cover can be opened；
(3) real tank sterilizing:
(3.1) medium sterilization:
Cover is fastened, interlayer heating is preheating to 90 DEG C, opens tank inside admission valve, when pressure inside the tank is raised to 0.05MPa, closes
Small exhaust steam valve, pressure rise to 0.11-0.12MPa, and temperature is 121-125 DEG C, keep the temperature 30 minutes；After medium sterilization, close each
Inlet valve (seeding tank connects sterilized racemosus pipeline under the protection of flames)；It is then turned on exhaust steam valve, is down to pressure inside the tank
When 0.02-0.03MPa, compressed air valve is opened, while cooling down is spare to 37 ± 1 DEG C；
(3.2) feed supplement liquid sterilizes:
Feed supplement tank steam valve is opened, interlayer heating when pressure rises to 0.05MPa, turns down exhaust steam valve, makes pressure inside the tank liter
To 0.11-0.12MPa, 121-125 DEG C of temperature, 45 minutes are kept the temperature；The sterilization pressure of 40% glucose feed supplement liquid is 0.09-
0.11MPa, temperature are 110-121 DEG C, keep the temperature 30 minutes；After sterilizing, each inlet valve is closed, opens exhaust steam valve and cooling
Valve is passed through microbiological free air when pressure is down to 0.02-0.03MPa, and be cooled to 37 ± 1 DEG C it is spare；
(4) sterilizing filter sterilizes:
While real tank sterilizes, the steam valve of sterilizing filter is opened, steam is made sufficiently to circulate, pressure 0.15-
0.20MPa, pressure maintaining 30 minutes, then pass to compressed air dry up 30 minutes or more it is spare；In addition the filter paper of sterilizing filter
Every weekly check replacement is primary；
Step 5) culture:
(1) it sows, culture transferring:
When temperature is down to 37 DEG C in seeding tank, by ready seed to be cultivated in sterile formality access seeding tank, to bacterium
Body grows into logarithmic growth phase, and when reaching a certain concentration (general 20,000,000,000/ml), takes kind and moves into and continues to train in fermentor
Support (sterilizing in 30-40 minutes before culture transferring of culture transferring pipeline is good spare)；
(2) it cultivates:
In incubation, temperature is observed at any time, foam in ventilatory capacity and tank, ventilatory capacity should be maintained at 0.05- when culture
0.08MPa, temperature should be maintained between 36-38 DEG C, pH value adjust every time after between 7.2-7.4, in the logarithmic growth of thallus
When the phase, ventilatory capacity will be increased accordingly；
(3) it samples:
Sow, before culture transferring with sterile formality (sample tap is led into steam sterilizing 15-20 minutes) sampling, detect the PH of culture medium
Value concentration, absorbs angle value, microscopy, and does steriling test；Inclined-plane is sowed 4 hours, and sample detection is started, and big bottle strain sows 2
Hour starts sample detection, 2 hours beginning sample detections after fermentor culture transferring, and detection pH value, bacterial type, absorbs angle value at concentration, often
Sub-sampling operator operates together with laboratory personnel, detects projects by lab technician, and result of laboratory test is notified operator in time
Member；Operator wants per half an hour to detect and adjusts pH value when thallus raised growth is bred；
(4) feed supplement:
In incubation, if foam can excessively take the circumstances into consideration that defoaming agent is added, according to the pH value concentration of detection, hydrogen after sterilizing is added
Sodium hydroxide solution makes pH value adjusted between 7.2-7.4, while adding suitable glucose solution；
Step 6) inactivation:
After the breeding of thallus raised growth, if concentration no longer obviously rises in 2-3 hours, pH value and absorption angle value are basically unchanged
In the case where, it is qualified through detection, formalin is added in 0.6% ratio, stirs 5-10 minutes, static 2 hours, thoroughly inactivates
Afterwards, precipitation operation personnel precipitation and separation is notified；
The preparation of step 7) flocculant:
(1) it dissolves:
Add water 780kg, interlayer is warming up to 40-60 DEG C, and polyacrylamide 150kg is added, and stirs 10-14 hours and dissolves；
(2) denaturation treatment:
After dissolution, add dimethylamine 28.8kg, formaldehyde 15kg, stirs 4-6 hours；
(3) it is acidified:
After denaturation treatment, with sulfuric acid tune pH value 3.8-4.1, stirring uses after 1 hour；
Step 8) precipitating, drying:
Fermentation liquid after inactivation is pressed into settling tank, between sulfuric acid tune pH value 6.0-6.5, in the ratio of 1.5-2.5%
Flocculant is added, stands 30 minutes after being thoroughly mixed, separates；Bacterium mud uniformly to be placed in baking pan, thickness is no more than 5cm,
85-95 DEG C 30-60 minutes dry, makes loss on drying≤6.0%；
Step 9) crushes:
By the fungus block after drying, first pass through 10 mesh coarse powder and be broken into little particle, then is finely divided, it is finely divided after bacterium powder should lead to
It sieves with 100 mesh sieve；
Step 10) mixing:
Bacterium powder is set in mixing machine, operates 8 minutes, bacterium powder is put into charging basket, the bacterium powder for finally mixing homogeneous is a batch；
Step 11) packaging:
Qualified bacterium powder will be examined by 20kg/ barrels of packagings, interior is double-layer plastic bag, is outside fiber drum, packing slip, bucket are put in bucket
Outer labelling, the name of an article, lot number, quantity should be consistent inside and outside bucket, conscientiously to check, be put in storage after the assay was approved.
Culture effect of the present invention is better than conventional culture medium, and low in cost, saves cost of material to pharmacy corporation,
Improve the added value of industry.Vitamin and other active materials will not be destroyed, nutritional ingredient balanced and reasonable is low in cost, can
Market is replaced often to use culture medium completely, fermentation efficiency also correspondinglys increase.
Specific embodiment
White staphylococcus powder of the present invention comprising following steps:
The preparation of step 1) strain:
(1) strain: staphylococcus albus, strain are upper white 71-1, and the long white 75-1 or white 72-1 in Hunan is 1-2 plants optional, by force
Chinese institute of biological products provides；
Bacterium colony: be creamy white colony on nutrient agar, and neat in edge is opaque, not chromogenesis；
Bacterial type: should be Gram-positive, in the coccus of botryoidalis irregular alignment；
(2) fungi preservation:
| Store method | Storage temperature | Holding time |
| Vacuum freezedrying preservation method | 0-4℃ | 3-5 |
| Atoleine preservation method | 0-4℃ | 6 months |
| Periodically transplanting preservation method | 0-4℃ | 1 month |
(3) breeding of strain:
Strain in use, when there is aging, degeneration and variation phenomenon, using Pure strain separation method in time to strain into
Row breeding, rejuvenation select more satisfactory strain for producing and using；
(4) preparation of culture medium:
The preparation of nutrient agar:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml, agar 15-20g；
It takes peptone and sodium chloride to be added in meat extract, after tepor dissolution, 15-20g agar is added and boils, it is molten that glucose is added
Solution, adjusting pH value make after sterilizing to be 7.2 ± 0.2, and packing, 121 DEG C sterilize 20 minutes, cold rear bevel；
The preparation of nutrient broth medium:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml；
Peptone, sodium chloride and glucose is taken to be added in meat extract, tepor dissolution, adjusting pH value makes after sterilizing to be 7.2 ± 0.2,
Packing, 121 DEG C sterilize 20 minutes, spare after cold；
(5) preparation of strain:
By sterility requirements, the strain in freeze-drying pipe is reached several test tube slants, 37 ± 1 DEG C culture 18-24 hours, it is cold
Hiding saves, and the strain of test tube slant is passed several flat bottle inclined-planes, 37 ± 1 DEG C culture 8-10 hours, use nutrient broth medium
Or uniform bacteria suspension is made for producing and using in physiological saline, can pass several test tube slants while passing flat bottle inclined-plane, cultivate
After save, and fill in strain passage record；
The digestion of step 2) basal liquid:
(1) dosage of basal liquid: corn pulp 450kg and water 900kg (or corn water 1600kg)；
(2) basal liquid digests: corn pulp and water (or corn water) being squeezed into digester with dnockout pumps, interlayer is warming up to
At 50-60 DEG C, with NaOH tune pH value to 8.0-8.5, steaming is continuously heating to 100 DEG C in tank, keeps the temperature 1 hour, at least places 10
Hour, cold water is cooled to 50 DEG C hereinafter, with hydrochloric acid tune pH value to 6.0-6.5 before filtering, is filtered with filter press spare；
Step 3) culture medium is prepared:
(1) in feed supplement bottle each composition of feed supplement liquid proportion, dosage and preparation:
| Title | With measuring bottle (kg) | Dose volume (ml) |
| 40% glucose | 3.2 | 8000 |
| 20% sodium hydroxide | 1.6 | 8000 |
| 10% organic silicon defoamer | 1.0 | 10000 |
It after above each composition precise, sufficiently dissolves, is poured into feed supplement bottle respectively, added with water to full dose, after wrapping
Sterilizing；
(2) in feed supplement tank each composition of feed supplement liquid proportion, dosage and preparation
After above each composition precise, respectively change in sugar bowl, choline tank and mixer after completely dissolution, with water mend to
Required volume is pumped into respectively in each feed supplement tank with vacuum pump, carries out real tank sterilizing, can also according to actual amount, according to the proportion into
Row is prepared；
(3) culture medium is prepared:
(3.1) proportion, dosage and preparation of seed tank culture base:
| Title | Corn pulp (corn water) digestive juice | Peptone | Sodium chloride | Oral glucose | Organic silicon defoamer | Total amount |
| Proportion | 15% (or 50%) | 1% | 0.2% | 0.3% | —— | —— |
| Dosage | 90kg (or 300kg) | 6.0kg | 1.2kg | 1.8kg | 200ml | 600000 ml |
Corn pulp (or corn water) digestive juice is squeezed into seeding tank, each material is added, is mended with water to full dose, stirring is abundant
After dissolution, after sodium hydroxide tune pH value to 7.4-7.6, sterilizing；
(3.2) proportion of fermentation tank culture medium, dosage and preparation:
| Title | Corn pulp (corn water) digestive juice | Peptone | Sodium chloride | Oral glucose | Organic silicon defoamer | Total amount |
| Proportion | >=15% (or 31.4%) | 1% | 0.2% | 0.3% | —— | —— |
| Dosage | >=525kg (or 1100kg) | 35.0kg | 7.0kg | 10.5kg | 1000ml | 3500000 ml |
Corn pulp (or corn water) digestive juice is squeezed into material-compound tank, each material is added, is mended with water to prescribed volume, stirring
After completely dissolution, with after sodium hydroxide tune pH value to 7.4-7.6, the culture medium after preparation is squeezed into fermentor with dnockout pumps,
Sterilizing；Step 4) sterilizing:
(1) in feed supplement bottle feed supplement liquid and required article sterilizing:
After feed supplement bottle and extraneous connector gauze, moistureproof paper wrapping, it is put into compact hand operated sterilizer, spare
Seed bottle, after racemosus pipeline is wrapped up with same method, while sterilizing；Sterilization pressure is 0.11-0.12MPa, temperature 121-
125 DEG C, 45 minutes 40% Glucose Liquids of heat preservation will individually sterilize, sterilization pressure 0.09-0.11MPa, temperature 110-121
DEG C, keep the temperature 30 minutes；
(2) slack tank sterilizes:
Just using tank or occurs microbiological contamination with tank, interruption and need to carry out slack tank sterilization treatment；Pre-temperature tank body, interlayer are warming up to first
It 90 DEG C, opens in tank with tank bottom inlet valve, allows steam to circulate in tank after twenty minutes, turn down exhaust steam valve, rise to 0.11- to pressure
0.12MPa, temperature are 121-125 DEG C, keep the temperature 45 minutes, after sterilizing, open exhaust steam valve, are reduced to zero to pressure inside the tank
When, cover can be opened；
(3) real tank sterilizing:
(3.1) medium sterilization:
Cover is fastened, interlayer heating is preheating to 90 DEG C, opens tank inside admission valve, when pressure inside the tank is raised to 0.05MPa, closes
Small exhaust steam valve, pressure rise to 0.11-0.12MPa, and temperature is 121-125 DEG C, keep the temperature 30 minutes；After medium sterilization, close each
Inlet valve (seeding tank connects sterilized racemosus pipeline under the protection of flames)；It is then turned on exhaust steam valve, is down to pressure inside the tank
When 0.02-0.03MPa, compressed air valve is opened, while cooling down is spare to 37 ± 1 DEG C；
(3.2) feed supplement liquid sterilizes:
Feed supplement tank steam valve is opened, interlayer heating when pressure rises to 0.05MPa, turns down exhaust steam valve, makes pressure inside the tank liter
To 0.11-0.12MPa, 121-125 DEG C of temperature, 45 minutes are kept the temperature；The sterilization pressure of 40% glucose feed supplement liquid is 0.09-
0.11MPa, temperature are 110-121 DEG C, keep the temperature 30 minutes；After sterilizing, each inlet valve is closed, opens exhaust steam valve and cooling
Valve is passed through microbiological free air when pressure is down to 0.02-0.03MPa, and be cooled to 37 ± 1 DEG C it is spare；
(4) sterilizing filter sterilizes:
While real tank sterilizes, the steam valve of sterilizing filter is opened, steam is made sufficiently to circulate, pressure 0.15-
0.20MPa, pressure maintaining 30 minutes, then pass to compressed air dry up 30 minutes or more it is spare；In addition the filter paper of sterilizing filter
Every weekly check replacement is primary；
Step 5) culture:
(1) it sows, culture transferring:
When temperature is down to 37 DEG C in seeding tank, by ready seed to be cultivated in sterile formality access seeding tank, to bacterium
Body grows into logarithmic growth phase, and when reaching a certain concentration (general 20,000,000,000/ml), takes kind and moves into and continues to train in fermentor
Support (sterilizing in 30-40 minutes before culture transferring of culture transferring pipeline is good spare)；
(2) it cultivates:
In incubation, temperature is observed at any time, foam in ventilatory capacity and tank, ventilatory capacity should be maintained at 0.05- when culture
0.08MPa, temperature should be maintained between 36-38 DEG C, pH value adjust every time after between 7.2-7.4, in the logarithmic growth of thallus
When the phase, ventilatory capacity will be increased accordingly；
(3) it samples:
Sow, before culture transferring with sterile formality (sample tap is led into steam sterilizing 15-20 minutes) sampling, detect the PH of culture medium
Value concentration, absorbs angle value, microscopy, and does steriling test；Inclined-plane is sowed 4 hours, and sample detection is started, and big bottle strain sows 2
Hour starts sample detection, 2 hours beginning sample detections after fermentor culture transferring, and detection pH value, bacterial type, absorbs angle value at concentration, often
Sub-sampling operator operates together with laboratory personnel, detects projects by lab technician, and result of laboratory test is notified operator in time
Member；Operator wants per half an hour to detect and adjusts pH value when thallus raised growth is bred；
(4) feed supplement:
In incubation, if foam can excessively take the circumstances into consideration that defoaming agent is added, according to the pH value concentration of detection, hydrogen after sterilizing is added
Sodium hydroxide solution makes pH value adjusted between 7.2-7.4, while adding suitable glucose solution；
Step 6) inactivation:
After the breeding of thallus raised growth, if concentration no longer obviously rises in 2-3 hours, pH value and absorption angle value are basically unchanged
In the case where, it is qualified through detection, formalin is added in 0.6% ratio, stirs 5-10 minutes, static 2 hours, thoroughly inactivates
Afterwards, precipitation operation personnel precipitation and separation is notified；
The preparation of step 7) flocculant:
(1) it dissolves:
Add water 780kg, interlayer is warming up to 40-60 DEG C, and polyacrylamide 150kg is added, and stirs 10-14 hours and dissolves；
(2) denaturation treatment:
After dissolution, add dimethylamine 28.8kg, formaldehyde 15kg, stirs 4-6 hours；
(3) it is acidified:
After denaturation treatment, with sulfuric acid tune pH value 3.8-4.1, stirring uses after 1 hour；
Step 8) precipitating, drying:
Fermentation liquid after inactivation is pressed into settling tank, between sulfuric acid tune pH value 6.0-6.5, in the ratio of 1.5-2.5%
Flocculant is added, stands 30 minutes after being thoroughly mixed, separates；Bacterium mud uniformly to be placed in baking pan, thickness is no more than 5cm,
85-95 DEG C 30-60 minutes dry, makes loss on drying≤6.0%；
Step 9) crushes:
By the fungus block after drying, first pass through 10 mesh coarse powder and be broken into little particle, then is finely divided, it is finely divided after bacterium powder should lead to
It sieves with 100 mesh sieve；
Step 10) mixing:
Bacterium powder is set in mixing machine, operates 8 minutes, bacterium powder is put into charging basket, the bacterium powder for finally mixing homogeneous is a batch；
Step 11) packaging:
Qualified bacterium powder will be examined by 20kg/ barrels of packagings, interior is double-layer plastic bag, is outside fiber drum, packing slip, bucket are put in bucket
Outer labelling, the name of an article, lot number, quantity should be consistent inside and outside bucket, conscientiously to check, be put in storage after the assay was approved.
Culture effect of the present invention is better than conventional culture medium, and low in cost, saves cost of material to pharmacy corporation,
Improve the added value of industry.Vitamin and other active materials will not be destroyed, nutritional ingredient balanced and reasonable is low in cost, can
Market is replaced often to use culture medium completely, fermentation efficiency also correspondinglys increase.
Although above having used general explanation, specific embodiment and test, the present invention is made to retouch in detail
It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Claims (2)
1. a kind of White staphylococcus powder, it is characterized in that including the following steps:
The preparation of step 1) strain:
(1) strain: staphylococcus albus, strain are upper white 71-1, and the long white 75-1 or white 72-1 in Hunan is 1-2 plants optional, are given birth to by Wuhan
Tetramune research institute provides；
Bacterium colony: be creamy white colony on nutrient agar, and neat in edge is opaque, not chromogenesis；
Bacterial type: should be Gram-positive, in the coccus of botryoidalis irregular alignment；
(2) fungi preservation:
Vacuum freezedrying preservation method, 0-4 DEG C of storage temperature, holding time 3-5；Atoleine preservation method, storage temperature 0-4
DEG C, the holding time 6 months；Periodically transplanting preservation method, 0-4 DEG C of storage temperature, the holding time 1 month；
(3) breeding of strain:
Strain in use, when there is aging, degeneration and variation phenomenon, in time selects strain using Pure strain separation method
It educates, rejuvenation, selects more satisfactory strain for producing and using；
(4) preparation of culture medium:
The preparation of nutrient agar:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml, agar 15-20g；
It takes peptone and sodium chloride to be added in meat extract, after tepor dissolution, 15-20g agar is added and boils, glucose dissolution is added, adjusts
Saving pH value makes after sterilizing to be 7.2 ± 0.2, and packing, 121 DEG C sterilize 20 minutes, cold rear bevel；
The preparation of nutrient broth medium:
Peptone 10g, sodium chloride 5g, glucose 1g, meat extract 1000ml；
Peptone, sodium chloride and glucose is taken to be added in meat extract, tepor dissolution, adjusting pH value makes after sterilizing to be 7.2 ± 0.2, packing,
121 DEG C sterilize 20 minutes, spare after cold；
(5) preparation of strain:
By sterility requirements, the strain in freeze-drying pipe is reached several test tube slants, 37 ± 1 DEG C culture 18-24 hours, refrigeration protect
Deposit, the strain of test tube slant passed several flat bottle inclined-planes, 37 ± 1 DEG C culture 8-10 hours, with nutrient broth medium or life
Uniform bacteria suspension is made for producing and using in reason salt water, can pass several test tube slants while passing flat bottle inclined-plane, protect after culture
It deposits, and fills in strain passage record；
The digestion of step 2 basal liquid:
(1) dosage of basal liquid: corn pulp 450kg and water 900kg (or corn water 1600kg)；
(2) basal liquid digests: corn pulp and water (or corn water) being squeezed into digester with dnockout pumps, interlayer is warming up to 50-60
DEG C when, with NaOH tune pH value to 8.0-8.5, steaming is continuously heating to 100 DEG C in tank, keeps the temperature 1 hour, at least place 10 hours,
Cold water is cooled to 50 DEG C hereinafter, with hydrochloric acid tune pH value to 6.0-6.5 before filtering, is filtered with filter press spare；
Step 3) culture medium is prepared:
(1) in feed supplement bottle each composition of feed supplement liquid proportion, dosage and preparation:
After 40% glucose, 20% sodium hydroxide and 10% organic silicon defoamer precise, sufficiently dissolves, pour into feed supplement bottle respectively
It is interior, it is added with water to full dose, is sterilized after wrapping；
(2) in feed supplement tank each composition of feed supplement liquid proportion, dosage and preparation:
After 40% glucose, 20% sodium hydroxide and 10% organic silicon defoamer precise, is changing sugar bowl, choline tank respectively and matching
In charging basket after completely dissolution, it is mended with water to required volume, is pumped into respectively with vacuum pump in each feed supplement tank, carry out real tank sterilizing,
It can be prepared according to the proportion according to actual amount；
(3) culture medium is prepared:
(3.1) proportion, dosage and preparation of seed tank culture base:
Corn pulp (or corn water) digestive juice is squeezed into seeding tank, each material is added, is mended with water to full dose, sufficiently dissolution is stirred
Afterwards, it with after sodium hydroxide tune pH value to 7.4-7.6, sterilizes；
(3.2) proportion of fermentation tank culture medium, dosage and preparation:
Corn pulp (or corn water) digestive juice is squeezed into material-compound tank, each material is added, is mended with water to prescribed volume, stirring is abundant
After dissolution, after sodium hydroxide tune pH value to 7.4-7.6, the culture medium after preparation is squeezed into fermentor with dnockout pumps, is sterilized；
Step 4) sterilizing:
(1) in feed supplement bottle feed supplement liquid and required article sterilizing:
After feed supplement bottle and extraneous connector gauze, moistureproof paper wrapping, it is put into compact hand operated sterilizer, spare kind
Sub- bottle after racemosus pipeline is wrapped up with same method, while sterilizing；Sterilization pressure is 0.11-0.12MPa, temperature 121-125
DEG C, 45 minutes 40% Glucose Liquids of heat preservation will individually sterilize, sterilization pressure 0.09-0.11MPa, and temperature is 110-121 DEG C, protect
Temperature 30 minutes；
(2) slack tank sterilizes:
Just using tank or occurs microbiological contamination with tank, interruption and need to carry out slack tank sterilization treatment；Pre-temperature tank body, interlayer are warming up to 90 first
DEG C, it opens in tank with tank bottom inlet valve, allows steam to circulate in tank after twenty minutes, turn down exhaust steam valve, rise to 0.11- to pressure
0.12MPa, temperature are 121-125 DEG C, keep the temperature 45 minutes, after sterilizing, open exhaust steam valve, are reduced to zero to pressure inside the tank
When, cover can be opened；
(3) real tank sterilizing:
(3.1) medium sterilization:
Cover is fastened, interlayer heating is preheating to 90 DEG C, opens tank inside admission valve, when pressure inside the tank is raised to 0.05MPa, the row of turning down
Steam valve, pressure rise to 0.11-0.12MPa, and temperature is 121-125 DEG C, keep the temperature 30 minutes；After medium sterilization, close respectively into vapour
Valve (seeding tank connects sterilized racemosus pipeline under the protection of flames)；It is then turned on exhaust steam valve, is down to 0.02- to pressure inside the tank
When 0.03MPa, compressed air valve is opened, while cooling down is spare to 37 ± 1 DEG C；
(3.2) feed supplement liquid sterilizes:
Feed supplement tank steam valve is opened, interlayer heating when pressure rises to 0.05MPa, turns down exhaust steam valve, rises to pressure inside the tank
0.11-0.12MPa, keeps the temperature 45 minutes by 121-125 DEG C of temperature；The sterilization pressure of 40% glucose feed supplement liquid is 0.09-
0.11MPa, temperature are 110-121 DEG C, keep the temperature 30 minutes；After sterilizing, each inlet valve is closed, opens exhaust steam valve and cooling
Valve is passed through microbiological free air when pressure is down to 0.02-0.03MPa, and be cooled to 37 ± 1 DEG C it is spare；
(4) sterilizing filter sterilizes:
While real tank sterilizes, the steam valve of sterilizing filter is opened, steam is made sufficiently to circulate, pressure 0.15-0.20MPa,
Pressure maintaining 30 minutes, then pass to compressed air dry up 30 minutes or more it is spare；In addition the every weekly check of the filter paper of sterilizing filter
Replacement is primary；
Step 5) culture:
(1) it sows, culture transferring:
It is raw to thallus by ready seed to be cultivated in sterile formality access seeding tank when temperature is down to 37 DEG C in seeding tank
It is long to enter logarithmic growth phase, and when reaching a certain concentration (general 20,000,000,000/ml), it takes kind and moves into fermentor and continue culture and (move
The sterilizing in 30-40 minutes before culture transferring of kind pipeline is good spare)；
(2) it cultivates:
In incubation, temperature is observed at any time, foam in ventilatory capacity and tank, ventilatory capacity should be maintained at 0.05- when culture
0.08MPa, temperature should be maintained between 36-38 DEG C, pH value adjust every time after between 7.2-7.4, in the logarithmic growth of thallus
When the phase, ventilatory capacity will be increased accordingly；
(3) it samples:
Sow, before culture transferring with sterile formality (sample tap is led into steam sterilizing 15-20 minutes) sampling, detect culture medium pH value,
Concentration absorbs angle value, microscopy, and does steriling test；Inclined-plane is sowed 4 hours, and sample detection is started, and big bottle strain is sowed 2 small
When start sample detection, 2 hours beginning sample detections after fermentor culture transferring, detection pH value, bacterial type, absorbs angle value at concentration, every time
Sampling operation personnel operate together with laboratory personnel, detect projects by lab technician, and result of laboratory test is notified operator in time；
Operator wants per half an hour to detect and adjusts pH value when thallus raised growth is bred；
(4) feed supplement:
In incubation, if foam can excessively take the circumstances into consideration that defoaming agent is added, according to the pH value concentration of detection, hydroxide after sterilizing is added
Sodium solution makes pH value adjusted between 7.2-7.4, while adding suitable glucose solution；
Step 6) inactivation:
After the breeding of thallus raised growth, if concentration no longer obviously rises in 2-3 hour, pH value and the feelings that are basically unchanged of absorption angle value
It is qualified through detection under condition, formalin is added in 0.6% ratio, stirs 5-10 minutes, static 2 hours, thoroughly after inactivation, leads to
Know precipitation operation personnel's precipitation and separation；
The preparation of step 7) flocculant:
(1) it dissolves:
Add water 780kg, interlayer is warming up to 40-60 DEG C, and polyacrylamide 150kg is added, and stirs 10-14 hours and dissolves；
(2) denaturation treatment:
After dissolution, add dimethylamine 28.8kg, formaldehyde 15kg, stirs 4-6 hours；
(3) it is acidified:
After denaturation treatment, with sulfuric acid tune pH value 3.8-4.1, stirring uses after 1 hour；
Step 8) precipitating, drying:
Fermentation liquid after inactivation is pressed into settling tank, between sulfuric acid tune pH value 6.0-6.5, wadding is added in the ratio of 1.5-2.5%
Solidifying agent, stands 30 minutes after being thoroughly mixed, and separates；Bacterium mud is uniformly placed in baking pan, thickness be no more than 5cm, 85-95 DEG C
It is 30-60 minutes dry, make loss on drying≤6.0%；
Step 9) crushes:
By the fungus block after drying, first pass through 10 mesh coarse powder and be broken into little particle, then is finely divided, it is finely divided after bacterium powder should pass through
100 meshes；
Step 10) mixing:
Bacterium powder is set in mixing machine, operates 8 minutes, bacterium powder is put into charging basket, the bacterium powder for finally mixing homogeneous is a batch；
Step 11) packaging:
Qualified bacterium powder will be examined by 20kg/ barrels of packagings, interior is double-layer plastic bag, is outside fiber drum, packing slip, bucket outer patch are put in bucket
Label, the name of an article, lot number, quantity should be consistent inside and outside bucket, conscientiously to check, be put in storage after the assay was approved.
2. White staphylococcus powder according to claim 1 is preparing the application in antitussive medicine.
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| CN108018249A (en) * | 2018-01-25 | 2018-05-11 | 仁和堂药业有限公司 | The processing of White staphylococcus powder and quality determining method |
| CN108048367A (en) * | 2018-01-25 | 2018-05-18 | 仁和堂药业有限公司 | White staphylococcus powder and its application |
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