CN104593280A - Probiotics medium and preparation method thereof - Google Patents
Probiotics medium and preparation method thereof Download PDFInfo
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- CN104593280A CN104593280A CN201310530558.6A CN201310530558A CN104593280A CN 104593280 A CN104593280 A CN 104593280A CN 201310530558 A CN201310530558 A CN 201310530558A CN 104593280 A CN104593280 A CN 104593280A
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- fungistat
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- 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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- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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Abstract
The invention relates to a novel medium capable of promoting rapid growth of probiotics, which belongs to the technical field of medical examination, and concretely relates to a probiotics medium and a preparation method thereof. The liquid medium comprises a basic medium, a growth stimulator, a nutrition agent and a bacterial depressant. The medium has the beneficial effect that the medium simultaneously contains nutrition component of the dual medium, rapid growth of probiotics can be promoted, and culture positive incidence is increased. According to the invention, Jerusalem artichoke powder is added as a tackifier and an active protective agent of probiotics, and activity of bacterium during a storage period can be ensured.
Description
Technical field
The present invention relates to a kind of novel culture medium that probiotic bacterium increases fast that can promote, belong to technical field of medical examination, specifically a kind of probiotic bacteria culture medium and preparation method thereof.
Background technology
Along with growth in the living standard, modern more payes attention to nutrition.But higher fatty acid, high protein is taken in too much, and foodstuff fibre is taken in very few, is easy to cause flora in body unbalance, thus causes state or even the disease of subhealth state.Regulated the microecological balance of human body by probiotic bacterium, while effectively preventing and treating various disease, make people away from Side effects of pharmaceutical drugs.Desire health long-lived a lot of people pursued, and the physical state that probiotic bacterium can help you to keep fit, eliminates healthy hidden danger.
Probiotic bacterium (Probiotics) is the living microorganism that a class is useful to host, is to be colonizated in human intestinal, reproductive system, can produce definite health efficacy thus improve host's microecological balance, plays the active beneficial microorganism general name of beneficial effect.Bacterium useful in human body, animal body or fungi mainly contain: clostridium butyricum, milk-acid bacteria, bifidus bacillus, Lactobacterium acidophilum, actinomycetes, yeast etc.The composite reactive probiotic bacterium of the most powerful product of the function studied in the world at present mainly above each quasi-microorganism composition, it is widely used in biotechnology, industrial or agricultural, food safety and life and health field.
In probiotic bacterium scale operation, the key of cost control improves the rate of loss of viable bacteria in microbial culture efficiency and the course of processing.The composition of the culture efficiency of probiotic bacterium and culture condition, substratum is closely related.Since the beginning of the nineties, panoramic " probiotic bacterium " class healthcare products are fashionable All Around The World.Meanwhile, the research of " probiotic bacterium " had become international popular research topic already.What is meant by probiotic bacterium? the universally recognized definition of international nutrition educational circles is: a kind of bacterium to animal health of prebiotic fungus strain, they can directly be taken as foodstuff additive, to maintain the balance of Intestinal flora.
Develop hundreds of probiotic bacterium healthcare product abroad, comprising: containing the yogurt milk of probiotic bacterium, yogourt, soybean milk yoghurt and oral liquid, tablet, capsule, powder agent, bacteriostasis spray etc. containing multiple probiotic bacterium.Few for people's research, the domestic cognition degree to probiotic bacterium is not also very strong, and this makes the product line of probiotic bacterium apply is not also very extensive, because survival rate is exactly a large technical barrier.
At present, the substratum generally used both at home and abroad, incubation time takes comparatively for a long time (usually needing 4 ~ 8 weeks), and positive rate not high (being generally 10 ~ 30%), far can not meet the needs of clinical diagnosis and treatment and diseases prevention.Therefore, in the urgent need to a kind of quick, sensitive probiotic bacteria culture medium.
Summary of the invention
The object of the present invention is to provide a kind of probiotic bacteria culture medium and preparation method thereof, based on traditional substratum, add liquid nutrient medium again and cultivate composition, this liquid nutrient medium comprises basic medium, growth promoter, nutrition agent and fungistat four kinds composition, this substratum is carefully selected materials in component and proportioning, make growth of probiotics health best in quality.
For achieving the above object, a kind of probiotic bacteria culture medium of the present invention, comprises basic medium, growth promoter, nutrition agent and fungistat four kinds composition, it is characterized in that: its composition and ratio is as follows:
Basic medium: copper sulfate 0.001 ~ 0.002g, zinc sulfate 0.001 ~ 0.002g, calcium chloride 0.001 ~ 0.002g, magnesium sulfate 0.01 ~ 0.02g, oleic acid 0.01 ~ 0.02g, Ferric Ammonium Citrate 0.05 ~ 0.1g, Sodium Citrate 0.1 ~ 0.2g, ammonium sulfate 0.2 ~ 0.4g, Sodium.alpha.-ketopropionate 0.5 ~ 1g, tween-80 0.5 ~ 1g, asparagine 1 ~ 2g, potassium primary phosphate 1 ~ 2g, SODIUM PHOSPHATE, MONOBASIC 2 ~ 4g, neutral glycerine 8 ~ 12ml, distilled water 1000ml.
Growth promoter: the former powder 150 ~ 300g of jerusalem artichoke, N.F,USP MANNITOL 20 ~ 40g, hydroxypropylmethylcellulose phthalate 50 ~ 200g, Vltra tears 20 ~ 30g, distilled water 1000ml.
Nutrition agent: amine DA-6 1 ~ 50g, emulsifying dispersant 1 ~ 5g, dispersion agent 2 ~ 10g, disintegrating agent 10 ~ 30g, distilled water 1000ml.
Fungistat: by concentration be respectively the penbritin of 50ug ~ lOOug/ml, amphotericin B, PXB, how pyridine ketone acid and azlocillin distilled water solution balanced mix form.
The component ratio of described basic medium, growth promoter, nutrition agent and fungistat is: basic medium 4 ~ 8ml, growth promoter 1 ~ 2ml, nutrition agent 8ml and fungistat 0.1 ~ 0.2ml.
Its preparation method of a kind of probiotic bacteria culture medium of the present invention, is characterized in that:
1, dissolved by basic medium each component distilled water, sterilizing 10 ~ 15min under 115 ~ 125 DEG C of high temperature, cooling, is placed in 4 DEG C of Refrigerator stores;
2, each component distilled water of growth promoter is dissolved, then sterilising filtration, packing, be placed in 4 DEG C of Refrigerator stores;
3, each component of fungistat is mixed with the solution that concentration is 50 ~ 1OOug/ml, then balanced mix with distilled water respectively;
4, finally basic medium, growth promoter and fungistat are added nutrition agent, the ingredient proportion of each component is: basic medium 4 ~ 8ml, growth promoter 1 ~ 2ml, nutrition agent 8ml, fungistat 0.1 ~ 0.2ml.
A kind of probiotic bacteria culture medium of the present invention and preparation method thereof, its beneficial effect is: the nutritive ingredient of this substratum simultaneously containing dual substratum, therefore can promote the quick growth of probiotic bacterium, improves and cultivates positive rate; Add jerusalem artichoke powder in the present invention as the increasing microbial inoculum of probiotic bacterium and activity protecting agent, ensure that the activity of lay up period bacterium.
Embodiment
Following examples are that the present invention goes out to set forth further.
Embodiment 1
1. basic medium moiety is as follows: copper sulfate 0.001g, zinc sulfate 0.001g, calcium chloride 0.001g, magnesium sulfate 0.01g, oleic acid 0.01g, Ferric Ammonium Citrate 0.05g, Sodium Citrate 0.1g, ammonium sulfate 0.2g, Sodium.alpha.-ketopropionate 0.5g, tween-80 0.5g, asparagine 1g, potassium primary phosphate 1g, SODIUM PHOSPHATE, MONOBASIC 2g, neutral glycerine 8ml, distilled water 1000ml; Compound method is first by above-mentioned each component dissolves, then 115 ~ 121 DEG C of autoclaving 10 ~ 15min, cooling, for subsequent use through the rearmounted 4 DEG C of Refrigerator stores of sterility test;
2. growth promoter moiety is as follows: pyridoxine hydrochloride 0.01g, vitamin H 0.01g, coenzyme A 0.1g, Triphosaden 0.1g, catalase 0.1g, a-naphthylacetic acid 0.1g, peptone 1g, glucose 10g, bovine serum albumin V5 factor 100g; Compound method is first dissolved by mentioned component, and then sterile filtration, packing, through the rearmounted 4 DEG C of Refrigerator stores of sterility test;
3. nutrition agent moiety is as follows: amine DA-6 1g, emulsifying dispersant 1g, dispersion agent 2g, disintegrating agent 10g, distilled water 1000ml; Compound method is first by above-mentioned each component dissolves, then 115 ~ 121 DEG C of autoclaving 10 ~ 15min, cooling, for subsequent use through the rearmounted 4 DEG C of Refrigerator stores of sterility test;
4. fungistat preparation: the distilled water solution respectively penbritin, amphotericin B, PXB, how pyridine ketone acid and azlocillin being mixed with 50ug ~ 1OOVg/ml concentration, gets balanced mix and forms.
5. get the liquid nutrient medium of above-mentioned assembly, wherein basis is cultivated 4ml, growth promoter 1ml, nutrition agent 4.9ml, fungistat 0.1ml, is mixed, and obtains required substratum.
Embodiment 2:
1. basic medium moiety is as follows: copper sulfate 0.0015g, zinc sulfate 0.0015g, calcium chloride 0.0016g, magnesium sulfate 0.018g, oleic acid 0.016g, Ferric Ammonium Citrate 0.07g, Sodium Citrate 0.15g, ammonium sulfate 0.3g, Sodium.alpha.-ketopropionate 0.7g, tween-80 0.7g, asparagine 1.3g, potassium primary phosphate 1.5g, SODIUM PHOSPHATE, MONOBASIC 3g, neutral glycerine 10ml, distilled water 1000ml; Compound method is by above-mentioned each composition water dissolution, then 115 ~ 121 DEG C of autoclaving 10 ~ 15min, cooling, for subsequent use through the rearmounted 4 DEG C of Refrigerator stores of sterility test;
2. growth promoter moiety is as follows: pyridoxine hydrochloride 0.015g, vitamin H 0.015g, coenzyme A 0.15g, Triphosaden 0.15g, catalase 0.15g, a-naphthylacetic acid 0.15g, peptone 1.5g, glucose 15g, bovine serum albumin V5 factor 150g; Compound method is dissolved by mentioned component with water, then sterile filtration, packing, through the rearmounted 4 DEG C of Refrigerator stores of sterility test; All the other conditions are with embodiment 1.
Claims (4)
1. a probiotic bacteria culture medium, comprises basic medium, growth promoter, nutrition agent and fungistat four kinds composition, it is characterized in that: its composition and ratio is as follows:
Basic medium: copper sulfate 0.001 ~ 0.002g, zinc sulfate 0.001 ~ 0.002g, calcium chloride 0.001 ~ 0.002g, magnesium sulfate 0.01 ~ 0.02g, oleic acid 0.01 ~ 0.02g, Ferric Ammonium Citrate 0.05 ~ 0.1g, Sodium Citrate 0.1 ~ 0.2g, ammonium sulfate 0.2 ~ 0.4g, Sodium.alpha.-ketopropionate 0.5 ~ 1g, tween-80 0.5 ~ 1g, asparagine 1 ~ 2g, potassium primary phosphate 1 ~ 2g, SODIUM PHOSPHATE, MONOBASIC 2 ~ 4g, neutral glycerine 8 ~ 12ml, distilled water 1000ml;
Growth promoter: the former powder 150 ~ 300g of jerusalem artichoke, N.F,USP MANNITOL 20 ~ 40g, hydroxypropylmethylcellulose phthalate 50 ~ 200g, Vltra tears 20 ~ 30g, distilled water 1000ml;
Nutrition agent: amine DA-6 1 ~ 50g, emulsifying dispersant 1 ~ 5g, dispersion agent 2 ~ 10g, disintegrating agent 10 ~ 30g, distilled water 1000ml;
Fungistat: by concentration be respectively the penbritin of 50ug ~ lOOug/ml, amphotericin B, PXB, how pyridine ketone acid and azlocillin distilled water solution balanced mix form.
2. a kind of probiotic bacteria culture medium as claimed in claim 1, is characterized in that: the component ratio of described basic medium, growth promoter, nutrition agent and fungistat is: basic medium 4 ~ 8ml, growth promoter 1 ~ 2ml, nutrition agent 8ml and fungistat 0.1 ~ 0.2ml.
3. an its preparation method for probiotic bacteria culture medium, is characterized in that:
1. dissolved by basic medium each component distilled water, sterilizing 10 ~ 15min under 115 ~ 125 DEG C of high temperature, cooling, is placed in 4 DEG C of Refrigerator stores;
2. each component distilled water of growth promoter is dissolved, then sterilising filtration, packing, be placed in 4 DEG C of Refrigerator stores;
3. each component of fungistat is mixed with the solution that concentration is 50 ~ 1OOug/ml, then balanced mix with distilled water respectively;
4. finally basic medium, growth promoter and fungistat are added nutrition agent mix obtained.
4. its preparation method of a kind of probiotic bacteria culture medium described in claim 3, is characterized in that: the ingredient proportion of described basic medium, growth promoter, fungistat, nutrition agent each component is: basic medium 4 ~ 8ml, growth promoter 1 ~ 2ml, nutrition agent 8ml, fungistat 0.1 ~ 0.2ml.
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| CN201310530558.6A CN104593280A (en) | 2013-10-31 | 2013-10-31 | Probiotics medium and preparation method thereof |
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| CN201310530558.6A CN104593280A (en) | 2013-10-31 | 2013-10-31 | Probiotics medium and preparation method thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101836629A (en) * | 2010-06-17 | 2010-09-22 | 陕西韦尔奇作物保护有限公司 | Plant growth regulator containing diethyl aminoethyl hexanoate |
| CN101843337A (en) * | 2010-06-07 | 2010-09-29 | 陕西科技大学 | Method for preparing probiotics tablets |
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2013
- 2013-10-31 CN CN201310530558.6A patent/CN104593280A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101843337A (en) * | 2010-06-07 | 2010-09-29 | 陕西科技大学 | Method for preparing probiotics tablets |
| CN101836629A (en) * | 2010-06-17 | 2010-09-22 | 陕西韦尔奇作物保护有限公司 | Plant growth regulator containing diethyl aminoethyl hexanoate |
Non-Patent Citations (3)
| Title |
|---|
| 孙天昊等: "干酪乳杆菌LC-STH-13高密度培养基优化", 《农产品加工》 * |
| 郭兴华: "《益生菌基础与应用》", 31 October 2002 * |
| 郭本恒: "《益生菌》", 31 January 2004 * |
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