CN1283782C - Photosynthetic bacteria culture medium and its use - Google Patents
Photosynthetic bacteria culture medium and its use Download PDFInfo
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- CN1283782C CN1283782C CN 200510012620 CN200510012620A CN1283782C CN 1283782 C CN1283782 C CN 1283782C CN 200510012620 CN200510012620 CN 200510012620 CN 200510012620 A CN200510012620 A CN 200510012620A CN 1283782 C CN1283782 C CN 1283782C
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- culture medium
- photosynthetic
- photosynthetic bacteria
- photosynthetic bacterium
- mistletoe
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Abstract
The present invention relates to a culture medium for photosynthetic bacteria. The culture medium is characterized in that every liter of the culture medium contains 2000 to 2.5g of coloed mistletoe herb extract measured by the original medicinal herb of the coloed mistletoe herb. The culture medium can reduce the growth period of the photosynthetic bacteria, increase the viable bacterium quantity of the photosynthetic bacteria and simultaneously eliminate the adherence phenomenon in the culturing process of the photosynthetic bacteria, and the culture medium is favorable for light to penetrate the wall of a container to accelerate the growth of the photosynthetic bacteria. The culture medium is suitable for the production of various photosynthetic bacterium preparations, for example, the culture medium can be used for the production of biofertilizer, feed additives and waste water purifiers from the photosynthetic bacteria and can also be used for the preparation of nanometer materials and antitumor drugs from the photosynthetic bacteria.
Description
Technical field
The present invention relates to a kind of microbiological culture media, specifically belong to a kind of photosynthetic bacteria culture medium and application thereof.
Background technology
(photosynthetic bacteria PSB) is the prokaryotic organism that a class has original luminous energy synthetic system to photosynthetic bacterium, and is very extensive in distributed in nature.Under different physical environments, have multiple functions such as fixed nitrogen, denitrogenation, solid carbon, sulfide oxidation.Since the eighties in last century, people progressively recognize the nutritive value of thalline and the physiologically active that some are special, be widely used in ecotope and human lives aspect, how in short period of time cheaply under the condition, obtain the highly active photosynthetic bacterial thallus of high density, produce actual needs to adapt to, the substratum of photosynthetic bacterium is a key factor.
The conventional substratum of a lot of photosynthetic bacteriums is disclosed in the prior art, as: the substratum that (1) University Of Shanxi photosynthetic bacterium research department uses: sodium acetate 1640mg, CaCl
2.2H
2O 75mg, MgSO
4.7H
2O 200mg, EDTA 20mg, yeast extract paste 1000mg, K
2HPO
4900mg, (NH
4)
2SO
41320mg, KH
2PO
4600mg, FeSO
4.7H
2O11.8mg, micro-1ml (wherein form: H by trace element
3BO
3280mg, Na
2MoO
4.2H
2O 75mg, CuSO
44mg, MnSO
4.4H
2O 210mg, ZnSO
4.7H
2O 24mg, deionized water 100ml), deionized water 1000ml, pH 6.8-7.2.
And for example: (2) Qian Cunrou, Huang Yixiu, " microbiology experiment study course " (BJ University Press, 2001:212) photosynthetic bacteria culture medium: NH of chief editor
4Cl 1000mg, NaHCO
31000mg, K
2HPO
4200mg, sodium acetate 1700mg, MgSO
4.7H
2O 200mg, NaCl 500mg, somatomedin 10ml (form: vitamins B by somatomedin
10.01mg the Buddhist nun restrains butyric acid 1mg, para-amino benzoic acid 1mg, vitamin H 0.01mg, deionized water 100ml), deionized water 1000ml, trace element solution 10ml (form: FeCl by trace element solution
3.6H
2O 5mg, CuSO
4.5H
2O 0.05mg, H
3BO
41mg, MnCl
2.4H
2O 0.05mg, ZnSO
4.7H
2O 1mg, Co (NO
3)
2.6H
2O 0.5mg, deionized water 1000ml), pH 6.8-7.2.
All there is following problem in various degree in these substratum: the photosynthetic bacterium growth cycle is long, and viable count is lower; Easily adherent in the photosynthetic bacterium growth process, light is not easy to see through container, influences photosynthetic bacterium growth.
Summary of the invention
The purpose of this invention is to provide and a kind ofly can effectively shorten the photosynthetic bacterium growth cycle, improve the photosynthetic bacterium vigor, increase the photosynthetic bacterium viable count, eliminate the photosynthetic bacteria culture medium of photosynthetic bacterium adherent phenomenon in process of growth.
A kind of photosynthetic bacteria culture medium provided by the invention is characterized in that with Herba Visci extract and water formulated.Contain Herba Visci extract in every liter of substratum by mistletoe crude drug 2000 grams-25 grams.
This substratum uses Herba Visci extract and water formulated fully, is suitable for utilizing photosynthetic bacterium to transform mistletoe and prepares in the antitumor drug and use.
Another kind of photosynthetic bacteria culture medium provided by the invention is characterized in that containing the conventional substratum of Herba Visci extract and photosynthetic bacterium.Contain Herba Visci extract in its every liter substratum by mistletoe crude drug 2000 grams-2.5 grams.
This photosynthetic bacteria culture medium not only can utilize photosynthetic bacterium to prepare bio-feritlizer, preparation fodder additives, prepare in the wastewater purificant and use, and also can utilize photosynthetic bacterium to prepare nano material, prepare in the antitumor drug and use.
Described Herba Visci extract is that mistletoe is pulverized, and with the pure liquid or the water of 7-30 times of volume of mistletoe crude drug quality, divides 2-4 time refluxing extraction 1-6 hour or decocting boiled 1-6 hour, reclaims alcohol in the extracting solution to there not being pure the flavor, and is concentrated into certain volume.Described mistletoe is the Loranthaceae mistletoe.Mistletoe is traditional Chinese medicine simply, and medicinal part is the exsiccant stem and branch with leaf.Its property is flat, and bitter has wind-damp dispelling, and invigorating the liver and kidney is nourished blood, and is antiabortive, hypotensive activity.Wherein contain flavonoid compound, alkaloid, triterpene, organic acid, phenylpropyl alcohol element, polypeptide, polysaccharide, protein, a small amount of sterol etc.Find that through test Herba Visci extract has nutrition to photosynthetic bacterium and promotes the effect of growth.
Described photosynthetic bacterium conventional substratum is meant that people often use in the prior art, and disclosed photosynthetic bacteria culture medium, as substratum cited in the background technology.
Advantage compared with prior art of the present invention and effect:
Substratum of the present invention not only can shorten the growth cycle of photosynthetic bacterium, improves the number of viable of photosynthetic bacterium; Simultaneously, can eliminate the adherent phenomenon in the photosynthetic bacterium culturing process, help light and see through wall of container, accelerate photosynthetic bacterium growth; This substratum is fit to the production of various photosynthetic bacteria preparations, can be used for photosynthetic bacterium and prepares bio-feritlizer, fodder additives, wastewater purificant, also can utilize photosynthetic bacterium to prepare nano material, prepare in the antitumor drug and use.
In addition, this patent has also been examined or check the influence of mistletoe to the photosynthetic bacterium dehydrogenase activity.Get the 2g wet thallus, with the polyvinyl alcohol is immobilization material, is fixing agent with saturated boric acid solution, makes immobilized cell 40g, after in conventional substratum, activating 12h, get 20g and be incubated in the conventional substratum of 250ml, 20g is incubated in the 250ml 25g mistletoe aqueous extract, after each cultivates 3d, discard nutrient solution, get immobilized cell with the distillation washed several times with water, measure absorbancy with triphenyltetrazolium chloride (TTC) method and characterize the photosynthetic bacterium dehydrogenase activity, the experiment triplicate.The immobilized cell absorbancy is 0.009 in the conventional substratum of photosynthetic bacterium, the immobilized cell absorbancy is 0.093 in the mistletoe extracting solution, be that the immobilized cell dehydrogenase activity improves more than 10 times than immobilized cell dehydrogenase activity in the conventional substratum in the mistletoe extracting solution, prove that mistletoe can improve the activity of photosynthetic bacterium.
Description of drawings
Fig. 1 is the influence of different concns mistletoe to the photosynthetic bacterium growth curve.
Among the figure: curve 1 is the growth curve of photosynthetic bacterium in the conventional substratum, and the about 20h of photosynthetic bacterium spends lag period, begins to enter exponential phase, and approximately 60h enters stationary phase, progresses into decline phase later on, and stationary phase, viable bacteria thalline content was 5.0 * 10
8Individual; Contain the growth curve of Herba Visci extract in the conventional substratum of curve 2 for 2 times of every liter of dilutions by photosynthetic bacterium in the nutrient solution of mistletoe crude drug 200g, photosynthetic bacterium is without lag period, directly enter exponential phase, approximately 40h enters stationary phase, and postpone to enter the time of decline phase, stationary phase, viable bacteria thalline content was 22.6 * 10
8Individual; Curve 3 contains the growth curve of Herba Visci extract by photosynthetic bacterium in the nutrient solution of mistletoe crude drug 12.5g in every liter of conventional substratum, photosynthetic bacterium is without lag period, directly enter exponential phase, approximately 50h enters stationary phase, and postpone to enter the time of decline phase, stationary phase, viable bacteria thalline content was 16.8 * 10
8Individual.Illustrate and contain the growth that Herba Visci extract is particularly suitable for photosynthetic bacterium in the substratum, can directly enter exponential phase, shortened the growth cycle of photosynthetic bacterium, and viable count is significantly improved without lag period.
Embodiment:
The invention will be further described below in conjunction with specific embodiment.
Embodiment 1: the photosynthetic bacteria culture medium experiment of Herba Visci extract and water preparation
With the stem and branch with leaf of mistletoe, behind natural air drying, pulverize.Mistletoe meal 2000g after the pulverizing divides 2 times with 40% ethanol 20000ml, each extraction 1 hour, merging filtrate, the ethanol in the reclaim under reduced pressure extracting solution, be concentrated into 1000ml, transfer pH to 7,121 ℃ of sterilization 30min are inoculated into photosynthetic bacterium seed liquor 60ml in the above-mentioned photosynthetic bacteria culture medium, under 1500Lux illumination, anaerobic condition, cultivated 5 days, and pressed the counting method of blood cell counting, the photosynthetic bacterium viable count is 42.6 * 10
8This embodiment illustrates that Herba Visci extract can be used as the perfect medium of photosynthetic bacterium growth.
Further to photosynthetic bacterium culture experiment under Herba Visci extract (by the mistletoe crude drug) different concns, the results are shown in Table 1 by this embodiment method.
Table 1 mistletoe different concns is to the influence (* 10 of photosynthetic bacterium growth viable count
8)
| Mistletoe is concentration (g/L) in substratum | 2000 | 1000 | 400 | 200 | 100 | 50 | 25 | 12.5 | 6.25 |
| The photosynthetic bacterium viable count | 42.6 | 40.4 | 28.3 | 21.5 | 12.6 | 8.7 | 3.6 | 1.4 | 0.2 |
Embodiment 2: contain the photosynthetic bacteria culture medium experiment of Herba Visci extract and conventional substratum
The Herba Visci extract preparation is with embodiment 1.Conventional substratum is with specification sheets (1) of the prior art substratum.With conventional substratum be diluted to the conventional substratum of 2 times, 4 times concentration with deionized water, be divided into 11 parts separately, and add Herba Visci extract make its separately concentration reach every liter 2000,1000,400,200,100,50,25,12.5,6.25,3.125 respectively, 0g (by the mistletoe crude drug), each sample placed 500 milliliters vial respectively, transfer pH to 7,121 ℃ of sterilization 30min, inoculate the photosynthetic bacterium seed liquor respectively, under 2000Lux illumination, anaerobic condition, cultivated 10 days, and pressed counting method of blood cell counting photosynthetic bacterium viable count.The results are shown in Table 2.
The conventional substratum of mistletoe of table 2 different concns and different concns is to the influence (* 10 of photosynthetic bacterium growth viable count
8)
As shown in Table 2, under conventional culture medium condition, along with reduce (2000-12.5g/L) of mistletoe concentration in the nutrient solution, the photosynthetic bacterium viable count is by 3.2 * 10
8Individually be increased to 16.8 * 10
8Individual, along with reduce (12.5-3.125g/L) of mistletoe concentration, the photosynthetic bacterium viable count is by 16.8 * 10 later on
8Individually reduce to 5.1 * 10
8Individual.Explanation is under conventional culture medium condition, when the mistletoe concentration ratio is low, mistletoe can promote the growth of photosynthetic bacterium, and be dose-effect relationship, but, begin to suppress the continued growth of photosynthetic bacterium to a certain extent along with the continuation of mistletoe concentration increases, when mistletoe concentration is 400g/L, fundamental sum does not have the conventional substratum viable count of Herba Visci extract identical, when concentration continues to increase, shows as more intense restraining effect; Be diluted under 2 times of conditions at conventional photosynthetic bacteria culture medium,, can promote the growth of photosynthetic bacterium always along with the increase of mistletoe extract concentration.Mistletoe concentration is during greater than 6.25g/L, the viable count 5.0 * 10 when viable count promptly begins to surpass conventional substratum
8Individual; Be diluted under 4 times of conditions at conventional photosynthetic bacteria culture medium,, can promote the growth of photosynthetic bacterium always along with the increase of mistletoe extract concentration.When mistletoe concentration during greater than 12.5g/L, the viable count 5.0 * 10 when bacterium liquid viable count begins greater than conventional substratum
8Individual, promptly Herba Visci extract can be used as the part substratum of photosynthetic bacterium growth.More than under the various culture condition, all do not have the adherent phenomenon of photosynthetic bacterium and take place.
Embodiment 3: the photosynthetic bacterium nutrient solution is to the restraining effect of mouse entity knurl Lewis lung cancer.
1. experiment material
The knurl strain: Lewis lung cancer, Chinese preventive medicine research institute provides, and University Of Shanxi goes down to posterity the photosynthetic bacterium research department and protects kind.Laboratory animal: C
57/ BL mouse, male and female half and half, body weight 18 ± 1.0g, Mountain Western Medicine S University's animal center provides.The contrast medicine: Cytoxan (CP), Hualian Pharmaceutical Co., Ltd., Shanghai produces, lot number: 0402061.
The photosynthetic bacterium nutrient solution, the photosynthetic bacterium nutrient solution of mistletoe concentration 1000g/L among the embodiment 1.
2. experimental technique
Get the fresh downright bad Lewis lung cancer tumor tissue that do not have, single cell suspension is made in homogenate under the aseptic condition, in 20 C
57The every mouse of/BL mouse right side armpit subcutaneous vaccination 0.2ml half-and-half is divided into 2 groups with mouse by male and female after 24 hours at random, 10 every group, establishes physiological saline control group, photosynthetic bacterium nutrient solution group.Give physiological saline control group every day mouse stomach administration physiological saline 1 time, dosage is 0.25ml/, give photosynthetic bacterium nutrient solution group every day mouse stomach administration 1 time, dosage press embodiment 1 photosynthetic bacterium nutrient solution 0.25ml/ (25g/kg * d), successive administration 15 days.Disconnected neck execution next day strips the knurl piece and weighs after the drug withdrawal, calculates tumour inhibiting rate by " tumour inhibiting rate=(1-test group knurl weight/physiological saline control group knurl is heavy) * 100% " formula.
3. experimental result: tumour inhibiting rate is 79.85%.
Embodiment 4: the application of photosynthetic bacterium nutrient solution in the celery plantation.
The photosynthetic bacterium nutrient solution that contains Herba Visci extract 12.5g/L with embodiment 2 made conventional substratum dilutes 12 times, flood seed and be degree seed soaking, young shoot plantation, growth stage 160 times of diluent foliage-sprays three times are arranged to seed, volume increase 40%, no insect pest takes place in the process of growth.
Embodiment 5: the application of photosynthetic bacterium nutrient solution in milk cattle cultivating.
The photosynthetic bacterium nutrient solution that contains Herba Visci extract 6.5g/L with embodiment 2 made conventional substratum adds feed with 1% ratio, mixes thoroughly and feeds, and also available atomizer evenly is sprayed in the feed feeds.Dilution is drunk 3ml/ day during drinking-water. and only, can increase the 4.6kg of giving milk every day, and the milk cow growth is fast, reduces sickness rate.
Embodiment 6: the application of photosynthetic bacterium nutrient solution in aquaculture.
The photosynthetic bacterium nutrient solution that contains Herba Visci extract 6.5g/L with embodiment 2 made conventional substratum is made following application test.
1. clean up the pond and spread at the end.Clean up the pond discharge water before, by every mu of 3kg consumption bacterium liquid and sandy soil are puddled, discharge water immediately after evenly being spread at the bottom of the pool.
2. the water surface is splashed.Every mu of water surface is evenly splashed in the water surface behind the bacterium liquid thin up with 3-5kg at every turn.Splashed 1 time in per 20 days, it is good that water quality keeps.
3. dipping bath is cured the disease.Bacterium liquid adds 20 times of dilutions of water, shrimp put into wherein, and dipping bath 10min, every day 1 time, for three days on end, the diseases prevention of being healthy and strong improves surviving rate.
Embodiment 7: the application of photosynthetic bacterium nutrient solution in percolate is handled.
It is centrifugal that conventional substratum is diluted to 4 times of photosynthetic bacterium nutrient solution 5000ml that contain Herba Visci extract 200g/L, makes the immobilization photosynthetic bacterium, is 72 hours at hydraulic detention time, and water inlet COD concentration is 8308mg/l, NH
3-N influent concentration is 1446mg/l, and the influent concentration of BOD is 2504mg/l, when the influent concentration of sulfide is 8.0mg/l, and, after 6 days dynamichandling, the clearance of sulfide is reached 97.5%, to NH
3The clearance of-N reaches 98.6%, and the clearance of BOD reaches 79.7%, and the clearance of COD reaches 82.6%.
Embodiment 8: the application of photosynthetic bacterium nutrient solution in the preparation nano PbS.
It is centrifugal that conventional substratum is diluted to 2 times of photosynthetic bacterium nutrient solution 1000ml that contain Herba Visci extract 400g/L, make the immobilization photosynthetic bacterium, after in conventional substratum, activating 12h, be incubated in the photosynthetic bacteria culture medium that 500ml contains Herba Visci extract, add the 300mg plumbic acetate simultaneously, co-cultivation gets nano level PbS 200mg.
Claims (3)
1, a kind of photosynthetic bacteria culture medium is characterized in that containing the conventional substratum of Herba Visci extract and photosynthetic bacterium.
2, the described photosynthetic bacteria culture medium of claim 1 is characterized in that containing in every liter of substratum Herba Visci extract by mistletoe crude drug 2000 grams-2.5 grams.
3, the described a kind of photosynthetic bacteria culture medium of claim 1 is utilizing photosynthetic bacterium to prepare application in bio-feritlizer, fodder additives, wastewater purificant, nano material, the antitumor drug.
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|---|---|---|---|
| CN 200510012620 CN1283782C (en) | 2005-06-18 | 2005-06-18 | Photosynthetic bacteria culture medium and its use |
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|---|---|---|---|
| CN 200510012620 CN1283782C (en) | 2005-06-18 | 2005-06-18 | Photosynthetic bacteria culture medium and its use |
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| CN1283782C true CN1283782C (en) | 2006-11-08 |
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| CN101602109B (en) * | 2009-06-15 | 2011-04-06 | 中北大学 | Preparation method for silver nano material |
| CN102224786B (en) * | 2011-04-28 | 2012-07-25 | 山西医科大学 | Method and application of photosynthetic bacteria culture solution in medicinal material planting |
| CN103409337A (en) * | 2013-07-01 | 2013-11-27 | 青岛浩澳环保科技有限公司 | Extracting raw material and extracting method of photosynthetic bacteria |
| CN103898022B (en) * | 2014-04-01 | 2017-01-25 | 湛江恒兴养殖技术服务有限公司 | Formula of photosynthetic bacteria culture medium and preparation method |
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Granted publication date: 20061108 Termination date: 20110618 |