CN1312273C - Additive for photosyntheric bacterial culture medium - Google Patents
Additive for photosyntheric bacterial culture medium Download PDFInfo
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- CN1312273C CN1312273C CNB021330743A CN02133074A CN1312273C CN 1312273 C CN1312273 C CN 1312273C CN B021330743 A CNB021330743 A CN B021330743A CN 02133074 A CN02133074 A CN 02133074A CN 1312273 C CN1312273 C CN 1312273C
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Abstract
The present invention relates to the culture of photosynthetic bacteria, particularly to an additive for a culture medium of the photosynthetic bacteria, and each liter of the culture medium contains 0.5 to 2g of sodium propionate and 10 to 100PPM of ferric ion salt. The product of the present invention can make the photosynthetic bacteria quickly propagate to form a photosynthetic bacterium product under suitable conditions for growth. The culture medium has low cost, wide range of application adaptability, high propagation speed of thalli and good production safety, and does not pollute ecological environment; the culture medium can be used in multiple fields of agriculture, environmental protection, medicine, etc. The photosynthetic bacteria are produced in large batches for being applied to agriculture, livestock and poultry industry and environmental protection, urban industrial organic sewage, industrial leftovers and agricultural rejected material are taken as raw materials after being comprehensively treated, the raw materials are inoculated with the photosynthetic bacteria for fermentation and culture, and a product with a high concentration of the photosynthetic bacteria is developed. When the method is applied to the industries of medicine, etc., the chemical reagents of analytical pure inorganic salt are taken as raw materials to produce a culture medium, and the culture medium is treated by inoculation, culture, purification and refinement to produce the photosynthetic bacterium product.
Description
Technical field
The present invention relates to the photosynthetic bacterium cultural method, specifically a kind of photosynthetic bacteria culture medium additive.
Technical background
The ecological distribution of photosynthetic bacterium (Photosynthetic Bacteria is called for short PSB) is very extensive, mainly is distributed in ocean, rivers, lakes and marhshes, active sludge and the soil, and be that a class is aquatic, the hydrosphere microorganism.Photosynthetic bacterium with sunlight as the energy, with multiple organism such as sodium-acetate, Sodium.alpha.-hydroxypropionate, N.F,USP MANNITOL, oxysuccinic acid, citric acid, glucose, glycerine is carbon source and nitrogenous source, and thalline has characteristics such as growth and breeding speed is fast, the substratum wide material sources are cultivated easily, meta-bolites is nutritious, and its product can be widely used in all many-sides such as fodder industry, agriculture production, environment protection; Therefore, more and more being subjected to scientific worker and business people payes attention to; In research in the past, people often pay attention to taxonomy status, molecular biology content, the photosynthetic fixed nitrogen of photosynthetic bacterium and put the process of hydrogen, photosynthetic fixed nitrogen and put in the hydrogen gene expression and regulation.In recent years, people are very extensive to the research of photosynthetic bacterium, applied research also obtains very big progress, many studies show that, photosynthetic bacterium all has higher using value at aspects such as agricultural environmental protection, medicine, after measured: photosynthetic bacterium dry mycelium protein content is 60.15%, and wherein composition of amino acid is reasonable; In addition, also contain required physiologically active substance of abundant VITAMIN and unsaturated fatty acids, bacterial polysaccharides, nucleic acid and various coenzyme and farm crop, animal and people's bulk-growth or the like.Be accompanied by the infiltration of market economy, people more and more pay attention to applied research and the utilisation technology exploitation of photosynthetic bacterium, photosynthetic bacterium are applied to aspects such as agricultural, Poultry farming industry, culture fishery, environment protection, soil sewage disposal and protective foods.But breeding that conventional its weak point of cultural method is strain growth is slow, bacteria containing amount is few, can not satisfy the requirement of production application far away.
Summary of the invention
The object of the present invention is to provide a kind of photosynthetic bacteria culture medium additive that can impel the quick growth and breeding of photosynthetic bacterium bacterial strain.
For achieving the above object, the technical solution used in the present invention is:
A kind of photosynthetic bacteria culture medium additive contains Sodium Propionate 0.5～2.0 gram in every liter of substratum, iron ion salt is 10～100PPM;
Wherein span is preferably: Sodium Propionate 0.6～1.5 gram/every liter of substratum, and iron ion salt is 40～100PPM; Better span is: Sodium Propionate 0.8～1.2 gram/every liter of substratum, and iron ion salt is 80～100PPM; Optimum value is: Sodium Propionate 1.0 gram/every liter of substratum, and iron ion salt is 100PPM;
Wherein said iron ion salt is organic iron ion salt (for example: ironic citrate, ferric ammonium citrate, ferric succinate, iron lactate etc.) or inorganic iron ion salt (for example: ferrous sulfate, iron(ic) chloride etc.);
Can also contain yeast extract paste 0.2～1.0 gram/every liter of substratum in the additive; Its preferably span be: 0.5～1.0 gram/every liter of substratum; Better span is: 0.8～1.0 gram/every liter of substratum; Optimum value is: 1.0 gram/every liter of substratum;
Described substratum be meant the laboratory use always city organic industrial sewage, agricultural byproducts waste or the industrial waste of seed fermentation substratum, comprehensive treating process in the usual way (for example: starch processing plant waste water tankage, bean-curd product factory waste water tankage or packing-house's waste water tankage) become can fermentation culture the nutrient raw material (substratum) of photosynthetic bacterium.
Using the cultural method that photosynthetic bacteria culture medium carries out the high density photosynthetic bacterium is: the photosynthetic bacterium seed liquor high amount of connecing (20～30%) is inoculated in the seed fermentation nutrient solution of expansion or is inoculated in the good fermentation culture of city organic industrial sewage, tankage comprehensive treating process, wherein add the photosynthetic bacteria culture medium additive, utilize natural lighting to make the photosynthetic bacterium breeding, every milliliter of photosynthetic bacterium is reached more than 2,000,000,000, and it is developed into the photosynthetic bacterium product.
Become photosynthetic bacterium seed liquor (seed liquor need be done the contrast indication with the standard bacterial classification and check to ensure the quality of products) according to the seed culture medium fermentation culture earlier at indoor or outdoors, seed culture is preferable composed as follows with fermention medium: ammonium chloride 1.0g, sodium-acetate 1.0g, sal epsom 0.2g, sodium bicarbonate 1.0g, sodium-chlor 0.5g, dipotassium hydrogen phosphate 0.2g, ironic citrate 0.005g, Sodium Propionate 1.0g, yeast extract paste 1.0g, peptone 1.0g, inorganic salts solution 10mL, growth cofactor 0.01g, distilled water 1000mL, PH=7.0.
The cofactor of wherein growing comprises: microbial culture such as VITMAIN B1, amino-benzene methyl alcohol, vitamin H are used material always.
Wherein inorganic salts solution is meant: the inorganic salts solution of trace, contain 5 milligrams of iron trichlorides, 0.05 milligram in calcium sulfate, 1 milligram of boric acid, 0.05 milligram of manganous chloride, 1 milligram in zinc sulfate in every liter of distilled water.
Require inoculum purity to want high, should note substratum sterilization and aseptic technique during operation.
Under laboratory condition, configuration seed fermentation substratum is a raw material with analytical pure inorganic salt chemical reagent and biochemical reagents, and the inoculation photosynthetic bacterium carries out fermentation culture.When producing photosynthetic bacterium in enormous quantities, be raw material after comprehensive treating process or be raw material, inoculate photosynthetic bacterium and carry out fermentation culture with analytical pure inorganic salt chemical reagent and biochemical reagents with city organic industrial sewage, industrial waste, agricultural byproducts waste.When the inoculation photosynthetic bacterium carries out fermentation culture, can utilize the nature sunlight to be the energy, can fluorescent lamp be the energy (perhaps in heliogreenhouse plastics booth) also, under 30～32 degree Celsius, cultivate, make its quick growth and breeding, photosynthetic bacterium can reach more than 2,000,000,000 for every milliliter, and it is developed into the photosynthetic bacterium product.
Can be first-selected during application to using in economic class crop such as vegetables, genseng and the fowl poultry kind animal.
The present invention has following advantage:
1. the photosynthetic bacterium breeding fast.Under identical adaptation growth conditions, to use the present invention and make the photosynthetic bacterium growth breeding fast, general collective media was cultivated fast 2～3 days, and the product bacterium is counted the concentration height, and general collective media is cultivated the bacterium number and is improved more than 30%.
2. culture medium raw material wide material sources.It is simply general that photosynthetic bacterium of the present invention is cultivated raw material; When producing photosynthetic bacterium in enormous quantities, can the city organic industrial sewage, industrial waste, agricultural byproducts waste be raw material after comprehensive treating process, inoculate and carry out fermentation culture behind the photosynthetic bacterium and be developed into high density photosynthetic bacterium product; Using at aspects such as pharmaceutical sectors, should be raw material with analytical pure inorganic salts chemical reagent then, makes substratum, inoculates, cultivates, purifies, makes the photosynthetic bacterium product.
3. purposes is wide.Can be used for all many-sides such as agricultural, environmental protection, medicine.
4. thalline adaptability is good, has reduced production cost.The present invention has reduced the requirement of photosynthetic bacterium to substratum nutrition, and production security is good, to eco-friendly.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment 1
Follow these steps to operation:
(1) plant female activation: the kind mother to photosynthetic bacterium activates (move and connect new inclined-plane, or be transferred to cultivate 3～5 days in the liquid nutrient medium); Wherein adopt the culture medium culturing that adds additive of the present invention to need 3 days to reach service requirements, adopt the culture medium culturing that does not add additive of the present invention then to need 5 days;
(2) seed fermentation is cultivated: the kind mother that photosynthetic bacterium is activated is linked in the seed fermentation substratum, (cultivate base unit weight is 70% to splendid attire in 2000 milliliters of transparent vials, surplus is an inoculum size), be placed on irradiation cultivation (temperature is at 20～30 ℃) under the nature sunlight, 7 days seed culture medium maturations, bacteria containing amount reaches per 200,000,000/ml, nutrient solution is scarlet or scarlet, the motion of test under microscope photosynthetic bacterium is active, and is neat, can use for enlarging fermentation culture;
The substratum of planting female activation and fermentation culture is:
Ammonium chloride 1.0g, sodium-acetate 1.0g, sal epsom 0.2g, sodium bicarbonate 1.0g, sodium-chlor 0.5g, dipotassium hydrogen phosphate 0.2g, ferrous sulfate 0.001g, Sodium Propionate 1.0g, yeast extract paste 1.0g, peptone 1.0g, growth cofactor (VITMAIN B1: amino-benzene methyl alcohol: vitamin H=1: 1: 1) 0.01g, inorganic salts solution 10mL, distilled water 1000mL, PH=7.0.
(3) enlarge fermentation culture: with photosynthetic bacterium seed fermentation nutrient solution with 30% big inoculum size, be encased in and enlarge in the fermentation culture, splendid attire (cultivating base unit weight is 70%) in 5000 milliliters of transparent vials (also can adopt the white plastic bucket) is placed on irradiation cultivation (in 20～30 ℃ of scopes of temperature) under the nature sunlight, enlarged the fermentation culture maturation in 10 days, bacteria containing amount reaches more than 1,000,000,000/milliliter, nutrient solution is scarlet or scarlet, the motion of test under microscope photosynthetic bacterium is active, the thalline stalwartness, pollution-free, free from extraneous odour, no abnormal phenomenon promptly can be applicable to the fertilisings of crops such as vegetables or genseng.Amount of application can be grasped and execute 1Kg photosynthetic bacterium expansion fermentation culture on per 100 square metres, and this nutrient solution can directly water root, also can spray rhizome base portions or blade face with 10 times of water dilution backs.
The compositing formula of this expansion fermention medium is as follows:
Ammonium chloride 1.0g, sodium-acetate 1.0g, sal epsom 0.2g, sodium bicarbonate 1.0g, sodium-chlor 0.5g, dipotassium hydrogen phosphate 0.2g, ferrous sulfate 0.001g, Sodium Propionate 1.0g, yeast extract paste 1.0g, peptone 1.0g, 10 milliliters of trace element solutions, 1000 milliliters in tap water
Embodiment 2
Can follow these steps to operation to the present invention:
(1) plants female activation: with embodiment 1 (1);
(2) seed fermentation is cultivated: with embodiment 1 (2);
(3) enlarge fermentation culture:
With photosynthetic bacterium seed fermentation nutrient solution (active photosynthetic bacterium number, every ml reaches 200,000,000), by 30% be transferred to enlarge expand in the fermention medium numerous, this expansion is selected soybean wastewater for use with fermention medium, the dress soybean wastewater is 7 kilograms in 10 kilograms of white plastic buckets, (consisting of of additive wherein: Sodium Propionate 1.0 grams/every liter, ferrous sulfate 0.001 gram/every liter, yeast extract paste 0.5 gram/every liter.) white plastic bucket after the switching can be placed under the nature solar irradiation and cultivate, 10 days maturations, its technical indicator can reach as following table 1.
Table 1
| PH | BOD(ppm) | COD(ppm) | The photosynthetic bacterium number (hundred million/ml) | |
| Before the cultivation | 6.4 | 7120 | 6840 | 0 |
| After the cultivation | 8.5 | 542 | 210 | 12 |
Embodiment 3
Can follow these steps to operation to the present invention:
(1) plants female activation: with embodiment 1 (1);
(2) seed fermentation is cultivated: with embodiment 1 (2);
(3) enlarge fermentation culture (under laboratory condition): inoculation and cultural method are with embodiment 1 (3);
Under laboratory condition, the fermention medium that adds additive is preferred, adorn 350 milliliters of non-additive substratum in 500 milliliters the bottle, the additive (its add-on is seen homogeneous design scheme table 2) that adds various combination, establish 9 processing altogether, be numbered 1,2,3,4,5,6,7,8,9 every then bottle graft kind photosynthetic bacterium seed liquor 30%, be placed under the nature solar irradiation and cultivated (25～30 ℃ of temperature) 10 days, measure photosynthetic bacterium bacterium number.
Table 2
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| Sodium Propionate (g/L) | 0 | 0.2 | 0.4 | 0.6 | 0.8 | 1.0 | 1.2 | 1.4 | 1.6 |
| Ironic citrate (g/L) | 0.002 | 0.006 | 0.01 | 0.014 | 0 | 0.004 | 0.008 | 0.012 | 0.016 |
| Yeast extract paste (g/L) | 0.3 | 0.2 | 0.1 | 0 | 0.35 | 0.25 | 0.15 | 0.05 | 0.4 |
Test is to organize in contrast with non-additive fermention medium;
Fermentative medium formula is composed as follows:
Ammonium chloride 1.0g, sodium-acetate 1.0g, sal epsom 0.2g, sodium bicarbonate 1.0g, sodium-chlor 0.5g, dipotassium hydrogen phosphate 0.2g, peptone 1.0g, inorganic salts solution 10mL, growth cofactor 0.01g, distilled water 1000mL, PH=7.0.The test-results such as the table 3 of different additive combination.
Table 3
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | Control group |
| Bacterium number (hundred million/milliliter) | 6.0 | 7.5 | 7.4 | 9.5 | 7.0 | 11.5 | 10.0 | 9.5 | 9.3 | 5.6 |
Control group is not contain the additive group in the fermention medium
Test-results shows, adds the best that is combined as of additive capacity (being Sodium Propionate 1.0 grams/every liter, ferrous sulfate 0.004 gram/every liter, yeast extract paste 0.25 gram/every liter) with No. 6 samples.
Embodiment 4
Can follow these steps to operation to the present invention:
(1) plants female activation: with embodiment 1 (1);
(2) seed fermentation is cultivated: with embodiment 1 (2);
(3), enlarge fermentation culture: photosynthetic bacterium seed fermentation nutrient solution (is contained the photosynthetic bacterium number more of living, every ml reaches 200,000,000), by 30% inoculum size be transferred to enlarge expand in the fermention medium numerous, this expansion fermention medium is selected for use and is butchered chicken house organic waste water, in 10 liters of white plastic buckets, be equipped with and butcher 7 kilograms of chicken house organic waste waters, (additive consists of: Sodium Propionate in interpolation again, ironic citrate, yeast extract paste), be placed on then and cultivate (20～30 ℃ of temperature) under the nature solar irradiation, 10 days, its photosynthetic bacterium content can reach as following table 4: every ml reaches 700,000,000, can use;
Table 4
| Handle numbering | Butcher chicken house waste water (Kg) | Sodium Propionate (g/L) | Ironic citrate (g/L) | Yeast extract paste (g/L) | Photosynthetic bacterium (hundred million/mL) |
| 1 | 5 | 2.0 | 0.01 | 1.0 | 9 |
| 2 | 5 | 1.0 | 0.005 | 0.5 | 8 |
| 3 | 5 | 0.5 | 0.001 | 0.25 | 5 |
| 4 | 5 | 0.25 | 0.0005 | 0.125 | 4.5 |
| 5 | 5 | 0.125 | 0.00025 | 0.0625 | 4 |
| 6 | 5 | 0.063 | 0.00013 | 0.0313 | 3 |
| 7 | 5 | 0 | 0 | 0 | 1.2 |
The all commercially available acquisition of the photosynthetic bacterium kind mother who is adopted in the embodiment of the invention there is no particular requirement to it.
The selection principle that enlarges fermention medium when the present invention uses is:
Use in agricultural, poultry industry, environmental protection industry, can the city organic industrial sewage, industrial waste, agricultural byproducts waste be raw material after comprehensive treating process, adds additive again; Using at aspects such as pharmaceutical sectors, should be raw material with analytical pure inorganic salts chemical reagent then, makes substratum, inoculation, cultivation.
(for example: photosynthetic bacterium 9218 bacterial classifications of Chinese Academy of Sciences's Shenyang preservation that ecological Studies separate), its effect is more remarkable for the photosynthetic bacterium bacterial classification of advantage for additive of the present invention.
Claims (9)
1. a photosynthetic bacteria culture medium additive is characterized in that: contain Sodium Propionate 0.5～2.0 gram in every liter of substratum, iron ion salt 10～16PPM.
2. according to the described photosynthetic bacteria culture medium additive of claim 1, it is characterized in that: wherein Sodium Propionate is 0.6～1.5 gram/every liter of substratum.
3. according to the described photosynthetic bacteria culture medium additive of claim 2, it is characterized in that: Sodium Propionate 0.8～1.2 gram/every liter of substratum wherein.
4. according to the described photosynthetic bacteria culture medium additive of claim 3, it is characterized in that: Sodium Propionate 1.0 gram/every liter of substratum wherein.
5. according to claim 1,2,3 or 4 described photosynthetic bacteria culture medium additives, it is characterized in that: wherein said iron ion salt is organic iron ion salt.
6. according to claim 1,2,3 or 4 described photosynthetic bacteria culture medium additives, it is characterized in that: wherein contain yeast extract paste 0.2～1.0 gram/every liter of substratum.
7. according to the described photosynthetic bacteria culture medium additive of claim 1, it is characterized in that: wherein said substratum is meant city organic industrial sewage, agricultural byproducts waste or the industrial waste of laboratory seed fermentation substratum commonly used or comprehensive treating process in the usual way.
8. a photosynthetic bacteria culture medium is characterized in that composed as follows: ammonium chloride 1.0g, sodium-acetate 1.0g, sal epsom 0.2g, sodium bicarbonate 1.0g, sodium-chlor 0.5g, dipotassium hydrogen phosphate 0.2g, ironic citrate 0.005g, Sodium Propionate 1.0g, yeast extract paste 1.0g, peptone 1.0g, inorganic salt solution 10mL, growth cofactor 0.01g, distilled water 1000mL, PH=7.0; Wherein said inorganic salt solution consists of: iron trichloride 5mg/L, calcium sulfate 0.05mg/L, boric acid 1mg/L, manganous chloride 0.05mg/L, zinc sulfate 1mg/L.
9. described photosynthetic bacteria culture medium Application of Additives of claim 1, it is characterized in that: with the photosynthetic bacterium seed liquor by weight 20～30% high amounts of connecing be inoculated in the substratum, wherein add the described photosynthetic bacteria culture medium additive of claim 1, utilize natural lighting to make the photosynthetic bacterium breeding.
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| CNB021330743A CN1312273C (en) | 2002-09-30 | 2002-09-30 | Additive for photosyntheric bacterial culture medium |
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| CNB021330743A CN1312273C (en) | 2002-09-30 | 2002-09-30 | Additive for photosyntheric bacterial culture medium |
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| CN1312273C true CN1312273C (en) | 2007-04-25 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102586371B (en) * | 2012-02-17 | 2014-01-01 | 中国科学院微生物研究所 | Method for producing swine Gp96 protein by using Pichia pastoris and special medium |
| US9555018B2 (en) * | 2013-03-15 | 2017-01-31 | Solenis Technologies, L.P. | Synergistic combinations of organic acid useful for controlling microoganisms in industrial processes |
| CN103243046B (en) * | 2013-05-02 | 2015-11-18 | 河北万林生物科技有限公司 | A kind of growth promoter and using method thereof impelling Photosynthetic bacterium strain Fast-propagation |
| CN112592852A (en) * | 2020-12-16 | 2021-04-02 | 中国热带农业科学院热带生物技术研究所 | Culture medium and application thereof in culture of marine photosynthetic bacteria |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993000443A1 (en) * | 1991-06-26 | 1993-01-07 | Bio-Technology General Corp. | Purification of recombinant apolipoprotein e from bacteria |
| JPH07184667A (en) * | 1993-12-24 | 1995-07-25 | Fuji Oil Co Ltd | Method for producing alcohols by using bacterium |
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2002
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993000443A1 (en) * | 1991-06-26 | 1993-01-07 | Bio-Technology General Corp. | Purification of recombinant apolipoprotein e from bacteria |
| JPH07184667A (en) * | 1993-12-24 | 1995-07-25 | Fuji Oil Co Ltd | Method for producing alcohols by using bacterium |
Non-Patent Citations (2)
| Title |
|---|
| 哈尔滨市污染水质中光合细菌的分离、鉴定及发酵条件研究 杨晨敏，哈尔滨师范大学自然科学学报，第15卷第6期 1999 * |
| 培养基组成对光合细菌产生类胡萝卜素的影响 肖念平，大连民族学院学报，第4卷第3期 2002 * |
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