CN1142265C - Bacterial strain resisting selenium, its preparing process and its application in preventing and treating arsenism - Google Patents
Bacterial strain resisting selenium, its preparing process and its application in preventing and treating arsenism Download PDFInfo
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Abstract
The present invention provides a strain with resistance to high selenium, which is prepared by the processes of primary selection, secondary selection and habituated culture of strains. The strain is high in bioactivity, and is greatly reduced in toxicity. The strain with resistance to high selenium can grow normally in a culture medium with 50PPM of selenium. The present invention also provides a preparing method of the strain with resistance to high selenium and the application of the strain with resistance to high selenium in medicines for preventing and curing arsenic poisoning.
Description
Technical field
The present invention relates to a kind of be used for the treatment of endemic microorganism and preparation method thereof and the application of this microorganism in the medicine of control arseniasis, specifically, the present invention relates to the application in the medicine of preparation control arseniasis of a kind of strain resisting selenium for the treatment of arseniasis and preparation method thereof and this strain resisting selenium.
Background technology
Endemic arsenic poisoning is popular in many countries, it is one of common multiple disease of serious threat human health, its harm not only shows as recent infringement to health, particularly serious is to cause that many physiological system damages of human body general and malignization at a specified future date change---canceration, sudden change and distortion, therefore the prevention and the treatment of arseniasis are paid much attention to by the World Health Organization and the Chinese government.
Find promptly that in China Taiwan the popular of endemic arsenic poisoning disease arranged the sixties, finds successively that after the eighties also there be the popular of endemic arsenic poisoning disease in provinces such as Xinjiang, the Inner Mongol, Guizhou, Shanxi, and the lesion constantly enlarges, and the state of an illness is on the rise.
At present, the method for treatment, control endemic arsenic poisoning disease roughly has two kinds, and the first is cut off high arsenic water source, changes the low arsenic water of drink.Facts have proved certain effect is arranged, can not get rid of Excessive Arsenic and reparation damaged tissue that human body is accumulated effectively though change the method for the low arsenic water of drink.In certain areas, need digging well to get water simultaneously, make that the fixture investment is bigger, under the present economic condition of China, be difficult to the assurance each department and can both bear so big burden economically; In addition, during owing to digging well to get water, the arsenic content in the motor-pumped well water is difficult to prediction, makes that digging well to get water operation easier in practice is bigger.Second method is to take medicine; At present China's first aid medicine that is used for heavy metal poisoning commonly used is a sulfhydryl compound, but the toxic side effect of sulfhydryl compound is bigger, and this medicine can not life-time service, and sign and damaged tissue are not had obvious repair.The real problem that goes out of another one is, sulfhydryl compound for endemic arsenic poisoning pernicious change in contingent long term all do not have preventive effect, and can only be used for acute treatment.
Selenium is the necessary trace element of human body and animal, the moiety of Selenoperoxidase, the imbalance of selenium content can cause some diseases in the human body, for example Keshan disease, Kaschin-Beck disease, cancer, cardiovascular and cerebrovascular disease, selenium deficiency, diabetes, the brown quality precipitation of matter, microcirculation disturbance, cataract or immunity and reproductive system underactivity etc.
Selenium and arsenic is toxinicide each other, and selenium has and suppresses and anticancer change, sudden change, distored pharmacology characteristic.But the inorganic selenium preparation is a violent in toxicity, and clinical application has certain danger.Therefore, seek that the efficient selenium preparation of low toxicity is applied to medicine industry and clinical treatment is the medicine scholar of many countries, the target that the chemist explores for many years always.
In sum, there is not the effective method of preventing and treating for endemic arsenic poisoning at present.Therefore, seek a kind of efficient, inexpensive, take medicine easy, the treatment endemic arsenic poisoning that is easy to apply, be a very urgent problem.Seeking a kind of microorganism that can be applied to treat endemic arsenic poisoning also becomes the problem of solution that the researchist presses for.
Summary of the invention
One of purpose of the present invention is to provide a kind of strain resisting selenium, and this bacterial classification has than high biological activity and its toxicity and reduces significantly, described strain resisting selenium can be in the seleno culture medium of 50PPM normal growth.
Two of purpose of the present invention is to provide the preparation method of described strain resisting selenium.Three of purpose of the present invention is to provide the application of described strain resisting selenium in the medicine of preparation control arseniasis.
To describe technical scheme of the present invention in detail below.
Selenium-resisting bacterium of the present invention is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), (abbreviation CGMCC) preservation at China Committee for Culture Collection of Microorganisms common micro-organisms center, preserving number is CGMCC NO.0454, and preservation date is on May 31st, 2000.
Strain resisting selenium of the present invention can be in the seleno culture medium of 50PPM normal growth, and can in the seleno matter of 50PPM, produce in batches, selenium content＞4000PPM is until reaching 6151PPM, this product zymic protein content＞38.0% in the product yeast.
Other physiological and biochemical indexs of strain resisting selenium of the present invention are identical with common medical yeast.
Strain resisting selenium of the present invention (yeast saccharomyces cerevisiae CGMCC NO.0454) makes by bacterial classification primary election, final election and domestication culturing process.
Strain resisting selenium of the present invention adopts following screening and cultivation program:
One, primary election bacterial classification
Primary election bacterial classification of the present invention can be selected from different types of yeast saccharomyces cerevisiae, comprising various cereuisiae fermentum or liquor yeast.Generally speaking, can select the primary election bacterial classification of 5-6 kind yeast saccharomyces cerevisiae for use as screening.
Two, substratum
Commodity maltose (rice or corn cerealose, 28 ℃, 39-40 ° of degree Beaume) is diluted to 6-7 ° of degree Beaume, adds 1 ‰ KH
2PO
4, 5 ‰ (NH
4)
2SO
4With the agar of 2-3%, after 1pt * 20min sterilization, add selenium solution, make the selenium concentration in the substratum reach required concentration, after shaking up, bevel or plane culture dish; Agar is wherein removed, promptly obtained fluid matrix.Described selenium solution can be used Na
2SeO
3Prepare with distilled water.
Three, screening procedure
(1) final election
The primary election bacterial classification of choosing is inoculated in respectively in the matrix of 150ml, on shaking table in 28 ℃, amplitude is to cultivate under 60-90 time/minute the condition to recover in 2 days, each bacterial classification that to recover then be seeded in respectively selenium content be 5,50,100,200,300,400 and the plating medium of 500PPM on, under 28 ℃ of-30 ℃ of conditions, cultivated 2-3 days, and chose wherein 2-3 the bacterial classification that on the higher substratum of selenium concentration, still can grow as the final election bacterial classification.
(2) domestication is cultivated
A. with the final election bacterial classification inoculation chosen in the plating medium of 10PPM selenium, under 28 ℃ of-30 ℃ of conditions, cultivated 3 days; Transfer to then in the identical plating medium of another new 10PPM, continue to cultivate 3 days with identical condition; And then transfer in the identical plating medium of another new 10PPM and cultivate, stable up to the bacterial classification growing way, till upgrowth situation is vigorous.
B. will transfer on the substratum that selenium content is 50PPM at the bacterial classification that growing way is stable on the 10PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
C. will transfer on the substratum that selenium content is 100PPM at the bacterial classification that growing way is stable on the 50PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
D. will transfer on the substratum that selenium content is 200PPM at the bacterial classification that growing way is stable on the 100PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
E. will transfer on the substratum that selenium content is 300PPM at the bacterial classification that growing way is stable on the 200PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
F. will transfer on the substratum that selenium content is 400PPM at the bacterial classification that growing way is stable on the 300PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
G. will transfer on the substratum that selenium content is 500PPM at the bacterial classification that growing way is stable on the 400PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
But h. be chosen under the selenium environment of 500PPM the still bacterial classification of normal growth, promptly obtain strain resisting selenium of the present invention.
Finish from 10PPM seleno culture medium domestication before transfer on the substratum that selenium content is 50PPM at final election bacterial classification of the present invention, can on being the substratum of 20PPM, 30PPM, 40PPM, selenium content repeat to tame step respectively once or once, progressively to improve the tolerance of final election bacterial classification to selenium.
Strain resisting selenium of the present invention on selenium-maltose agar or the selenium-wort agar substratum after cultivating 2-3 days on 28 ℃-30 ℃, can see orange red, circular projection bacterium colony, surface wettability, thickness, and distributed the sweet-smelling smell, showing that this bacterial classification has can make low sugar be converted into the physiological characteristic of alcohol and carbonic acid gas, becomes irregular cycle under mirror.Strain resisting selenium of the present invention removes can be at the above normal growth of selenium content 50PPM, and be orange red outside, other physiology, biochemical characteristic are all identical with common yeast saccharomyces cerevisiae.
Screening procedure of the present invention is that the biologically biological principle that environment is progressively adapted to of basis designs; In each culture cycle,, make barms progressively obtain the characteristic of anti-high selenium through many generation breedings; Along with the raising of environment selenium concentration, the prolongation of incubation time, the anti-selenium characteristic of bacterial classification is also stable day by day.Screening process of the present invention is through a plurality of selenium concentration gradients, cultivated in many days, forms new strain resisting selenium at last, and is applied to industrial production.
Strain resisting selenium of the present invention can being applied in the medicine of control arseniasis.
The selenium content numerical value of the selenium yeast that strain resisting selenium of the present invention is produced in different seleno matter is listed in table 1.
Table 1
| Selenium content in the matrix (PPM) | Incubation time and selenium content (PPM) | |
| 24 hours | 72 hours | |
| 20 | 1474 | 1965 |
| 30 | 2473 | 3126 |
| 40 | 3681 | 4727 |
| 50 | 4472 | 6151 |
Experiment showed, that strain resisting selenium of the present invention has special anti-selenium, the absorption of the inorganic selenium increase with selenium content in the culture medium is increased; The selenium content and the selenium concentration in the matrix of the selenium yeast of producing as can be seen, are significant proportional relation.
Inorganic selenium content how much be one of key factor that influences the selenium yeast quality product, can adopt method as described below to measure inorganic selenium content in the selenium yeast: in centrifuge tube, to add finished product selenium yeast and proper amount of deionized water, shook 5 minutes, centrifugal, get supernatant liquor, measure its solubility selenium content.
The form of Selenium in Selenium-Yeast is directly connected to the biologic activity of selenium.About not knowing its definite component and structure before the form-order of Selenium in Selenium-Yeast as yet fully.Classic methods is a dialysis method at present.
Table 2 is the result with 0.1N NaOH dialysis selenium yeast.
Table 2
| Total selenium before the dialysis | Selenium in the yeast of dialysis back | The selenium that gives | Dialysance (%) |
| 4063.78 | 2365.50 | 1703.28 | 41.91 |
| 4126.28 | 2365.50 | 1760.78 | 42.66 |
| 4169.60 | 2433.26 | 1736.34 | 41.64 |
| 4121.56±50.57 | 2388.08±39.12 | 1733.47±28.86 | 42.06 |
The result of table 2 shows, is retained in the selenium about 58% in the selenium yeast.This part selenium mainly is the protein of methionine(Met) form or the organoselenium of other macromole attitudes, and the selenium of dialysis then is organic selenium and a small amount of unassimilated inorganic states selenium of small molecular weights such as the selenoamino acid, selenium gsh of non-combined.
Table 3 is the animal acute toxicity experiment result of selenium yeast of the present invention.
Table 3
| Group | Dosage (g/kg) | Log10 dose | Animal dead number (only) | Mortality ratio (%) |
| 1 | 16.443 | 1.2160 | 20/20 | 100 |
| 2 | 13.982 | 1.1460 | 19/20 | 95 |
| 3 | 11.890 | 1.0750 | 7/20 | 35 |
| 4 | 10.110 | 1.0050 | 4/20 | 20 |
| 5 | 8.587 | 0.9340 | 2/20 | 10 |
| 6 | 7.310 | 0.8630 | 0/20 | 0 |
Can learn its medium lethal dose of selenium yeast of the present invention LD by table 3
50Be 11.72 ± 0.5g/kg, as calculated, amounting to pure selenium is 62.668mg/kg, and the minimum lethal dose of selenium yeast is 8.60g/kg.The LD of the present inorganic states Sodium Selenite of using always
50Be 8.77-14.560mg/kg, the toxicity of selenium yeast only is 1/4 of Sodium Selenite well below Sodium Selenite, therefore, can guarantee the security of clinical use.
Through experimentation on animals, selenium yeast of the present invention all influences less than obvious free activity, blood pressure, heart rate, respiratory rate and the degree of depth of animal.Selenium yeast does not have mutagenesis to organism, does not cause the rising of chromosome aberration frequency, does not also bring out micronuclear rates and raises.
Human body is listed in table 4 relative bioavailability of selenium yeast
Table 4
| Group | Project | Send out selenium (μ g/g) | Blood selenium (μ g/ml) |
| Sodium Selenite | Selenium concentration before the clothes selenium | 0.060 | 0.010 |
| Clothes selenium concentration in the time of three months | 0.195 | 0.039 | |
| Increase concentration behind the clothes selenium | 0.135 | 0.029 | |
| Relative utilization ratio (%) | 100 | 100 | |
| The selenium yeast group | Selenium concentration before the clothes selenium | 0.062 | 0.0099 |
| Clothes selenium concentration in the time of three months | 0.318 | 0.061 | |
| Increase concentration behind the clothes selenium | 0.256 | 0.052 | |
| Relative utilization ratio (%) | 189.63 | 179.31 |
Being used for the treatment of in the endemic arsenic poisoning process comparable situation of sign and clinical symptom comprehensive therapeutic effect lists in table 5.
Table 5
| Handle | The example number | Effectively | Constant | Worsen | Effectively-worsen | ||||
| The example number | ％ | The example number | ％ | The example number | ％ | The example number | ％ | ||
| Selenium treatment group | 100 | 88 | 88.00 | 7 | 7.00 | 4 | 4.00 | 1 | 1.00 |
| Control group | 86 | 27 | 31.39 | 35 | 40.70 | 13 | 15.12 | 11 | 12.79 |
List in table 6 for the comparable situation that sign and clinical symptom in the treatment endemic arsenic poisoning process lapse to.
Table 6
| Handle | S or S | The example number | Take a turn for the better | Constant | Worsen | Improvement-deterioration | ||||
| The example number | ％ | The example number | ％ | The example number | ％ | The example number | ％ | |||
| Selenium treatment group | Sign | 100 | 75 | 75.00 | 20 | 20.00 | 4 | 4.00 | 1 | 1.00 |
| Symptom | 100 | 55 | 55.00 | 45 | 45.00 | 0 | 0.00 | 0 | 0.00 | |
| Control group | Sign | 86 | 22 | 25.58 | 52 | 60.47 | 9 | 10.47 | 3 | 3.49 |
| Symptom | 86 | 21 | 21.41 | 51 | 59.30 | 13 | 15.12 | 1 | 1.12 | |
Being used for the treatment of the comparable situation that the liver function positive lapses in the endemic arsenic poisoning process lists in table 7.
Table 7
| Handle | Positive routine number | Turn out cloudy | Positive | ||
| The example number | ％ | The example number | ％ | ||
| Selenium treatment group | 25 | 20 | 80.00 | 5 | 20.00 |
| Control group | 13 | 6 | 46.15 | 7 | 53.85 |
Being used for the treatment of the comparable situation that the liver B ultrasonic lapses to unusually in the endemic arsenic poisoning process lists in table 8.
Table 8
| Handle | The example number | Recovery from illness | Take a turn for the better | Total effective rate | Constant | Increase the weight of | |||||
| The example number | ％ | The example number | ％ | The example number | ％ | The example number | ％ | The example number | ％ | ||
| Selenium treatment group | 25 | 8 | 32.00 | 7 | 28.00 | 15 | 60.00 | 10 | 40.00 | 0 | 0.00 |
| Control group | 13 | 1 | 7.60 | 3 | 23.08 | 4 | 30.77 | 3 | 23.08 | 6 | 46.15 |
Being used for the treatment of the comparable situation that electrocardiogram(ECG lapses to before and after the treatment in the endemic arsenic poisoning process lists in table 9:
Table 9
| Handle | Before the treatment | After the treatment | |||||||||
| The state of an illness | Total routine number | Unusually | Normally | Unusually | Normally | ||||||
| Take a turn for the better | ％ | Constant | ％ | Worsen | ％ | Normally | Worsen | ||||
| Selenium treatment group | 97 | 46 | 51 | 39 | 84.8 | 6 | 13.0 | 1 | 2.2 | 51 | 0 |
| Control group | 63 | 29 | 34 | 13 | 44.8 | 13 | 44.8 | 4 | 13.8 | 34 | 0 |
Being used for the treatment of the comparable situation that the nail fold microcirculation changes before and after the treatment in the endemic arsenic poisoning process lists in table 10.
Table 10
| Handle | The example number | Take a turn for the better | ％ | No change | ％ | Worsen | ％ |
| Clothes selenium group | 94 | 53 | 56.38 | 39 | 41.49 | 2 | 2.13 |
| Control group | 61 | 22 | 36.07 | 33 | 54.10 | 6 | 9.84 |
The comparable situation that is used for the treatment of treatment front and back red corpuscle electron microscopic observation result in the endemic arsenic poisoning process is listed in table 11.
Table 11
| Handle | The example number | Repair and take a turn for the better | ％ | No change | ％ | Worsen | ％ |
| Clothes selenium group | 18 | 13 | 72.22 | 5 | 27.78 | 0 | 0 |
| Control group | 18 | 0 | 0 | 10 | 55.56 | 8 | 44.44 |
Being used for the treatment of in the endemic arsenic poisoning process the dynamic comparable situation of whole blood GSHPX lists in table 12.
Table 12 (unit: unit of activity)
The comparable situation that is used for the treatment of treatment front and back SOD enzymic change in the endemic arsenic poisoning process is listed in table 13.
Table 13 (unit: ng/mg.Hb)
The dynamic change situation that is used for the treatment of blood in the endemic arsenic poisoning process, urine, a middle arsenic, selenium is listed in table 14.
Table 14
| Handle and the course of treatment | Before the test | Tested back 3 months | Tested back 9 months | Tested back 14 months | 1,4 clean liftings (%) | |
| Selenium treatment group | Blood arsenic (μ g/ml) | 0.070±0.099 | 0.072±0.099 | 0.024±0.010 | 0.016±0.006 | -77.14 |
| Urine arsenic (μ g/ml) | 0.255±0.306 | 0.195±0.163 | 0.106±0.163 | 0.030±0.030 | -88.24 | |
| Send out arsenic (μ g/g) | 2.970±1.627 | 2.960±2.627 | 1.659±1.495 | 0.798±0.603 | -73.13 | |
| Control group | Blood arsenic (μ g/ml) | 0.101±0.216 | 0.046±0.039 | 0.031±0.016 | 0.027±0.011 | -73.27 |
| Urine arsenic (μ g/ml) | 0.381±0.535 | 0.210±0.262 | 0.092±0.088 | 0.053±0.045 | -86.10 | |
| Send out arsenic (μ g/g) | 2.824±2.264 | 2.910±2.520 | 1.932±1.552 | 1.480±1.447 | -47.59 | |
| Selenium treatment group | Blood selenium (μ g/ml) | 0.128±0.029 | 0.157±0.030 | 0.158±0.018 | 0.181±0.025 | +41.41 |
| Urine selenium (μ g/100ml) | 1.554±0.779 | 2.293±1.167 | 2.867±1.128 | 2.955±1.792 | +90.15 | |
| Send out selenium (μ g/g) | 0.446±0.061 | 0.499±0.074 | 0.546±0.069 | 0.560±0.075 | +25.56 | |
| Control group | Blood selenium (μ g/ml) | 0.129±0.026 | 0.133±0.022 | 0.137±0.016 | 0.131±0.014 | +1.55 |
| Urine selenium (μ g/100ml) | 2.051±1.290 | 1.717±0.679 | 2.085±0.845 | 1.783±0.808 | -13.07 | |
| Send out selenium (μ g/g) | 0.477±0.068 | 0.465±0.055 | 0.477±0.081 | 0.475±0.056 | -0.42 |
Use the selenium content in the selenium yeast that preparation method of the present invention obtains to be approximately 4000PPM.
Compared with prior art, the present invention has following outstanding substantive distinguishing features:
1. the present invention utilizes modern biotechnology that inorganic selenium is converted into organic selenium compounds its toxicity is significantly reduced, and biological activity is greatly improved.
2. organoselenium zymic production technique of the present invention is simple, cheap.
3. the organoselenium yeast not only can be used for the treatment of arsenicalism, can also be applied to prevent because the malignization at a specified future date that arsenicalism causes changes; That is to say that this selenium yeast not only has the chemotherapy effect, also has the chemoprophylaxis effect.
4. present, common bacterial classification can only be produced under the condition of 5PPM, and the selenium content that is produced in the yeast is about 500PPM; And selenium yeast of the present invention is produced under the condition of 50PPM, and the organic selenium content in the yeast surpasses 95%, and selenium content can reach more than the 4000PPM, therefore, can significantly reduce the consumption of selenium yeast raw material in the finished product preparation, has also just reduced production cost.
Embodiment:
Describe the present invention further below by embodiment, the present invention is more in depth understood with promotion.But the present invention is not subjected to the restriction of these embodiment.Those skilled in the art can make amendment or improve under the guide of spirit of the present invention the present invention, all belong to apparent.Protection scope of the present invention proposes in claims.
Embodiment 1
Select for use yeast1419,1571,1422,1603,1328,1599 as the primary election bacterial classification available from Beijing nutrition source institute.
Aforesaid bacterial classification is inoculated in respectively in the matrix of 150ml, on shaking table in 28 ℃, amplitude is to cultivate under 80 times/minute the condition to recover in 2 days, each bacterial classification that to recover then be seeded in respectively selenium content be 5,50,100,200,300,400 and the plating medium of 500PPM on, under 30 ℃ of conditions, cultivated 3 days, and chose wherein 3 bacterial classifications that on the higher substratum of selenium concentration, still can grow as the final election bacterial classification.
A. the final election bacterial classification inoculation that true good general chooses was cultivated 3 days under 30 ℃ of conditions in the plating medium of 10PPM selenium; Transfer to then in the identical plating medium of another new 10PPM, continue to cultivate 3 days with identical condition; And then transfer in the identical plating medium of another new 10PPM and cultivate, stable up to the bacterial classification growing way, till upgrowth situation is vigorous.
B. will transfer on the substratum that selenium content is 50PPM at the bacterial classification that growing way is stable on the 10PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
C. will transfer on the substratum that selenium content is 100PPM at the bacterial classification that growing way is stable on the 50PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
D. will transfer on the substratum that selenium content is 200PPM at the bacterial classification that growing way is stable on the 100PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
E. will transfer on the substratum that selenium content is 300PPM at the bacterial classification that growing way is stable on the 200PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
F. will transfer on the substratum that selenium content is 400PPM at the bacterial classification that growing way is stable on the 300PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
G. will transfer on the substratum that selenium content is 500PPM at the bacterial classification that growing way is stable on the 400PPM seleno culture medium and cultivate 3 days; As the method for a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable.
But h. be chosen under the selenium environment of 500PPM the still bacterial classification of normal growth, promptly obtain strain resisting selenium of the present invention.
Claims (4)
1. a S. cervisiae (Saccharomyces cerevisiae), its preserving number is CGMCCNO.0454.
2. the preparation method of the described S. cervisiae of claim 1 is characterized in that:
Described yeast saccharomyces cerevisiae bacterial classification makes by bacterial classification primary election, final election and domestication culturing process;
The bacterial classification of described bacterial classification primary election process is selected cereuisiae fermentum or liquor yeast for use;
Described final election process is: the primary election bacterial classification of choosing is inoculated in respectively in the matrix of 150ml, on shaking table in 28 ℃, amplitude is to cultivate under 60-90 time/minute the condition to recover in 2 days, each bacterial classification that to recover then be seeded in respectively selenium content be 5,50,100,200,300,400 and the plating medium of 500PPM on, under 28 ℃ of-30 ℃ of conditions, cultivated 2-3 days, and chose wherein 2-3 the bacterial classification that on the higher substratum of selenium concentration, still can grow as the final election bacterial classification;
Described domestication culturing process is:
A. with the final election bacterial classification inoculation chosen in the plating medium of 10PPM selenium, under 28 ℃ of-30 ℃ of conditions, cultivated 3 days; Transfer to then in the identical plating medium of another new 10PPM, continue to cultivate 3 days with identical condition; And then transfer in the identical plating medium of another new 10PPM and cultivate, stable up to the bacterial classification growing way, till upgrowth situation is vigorous;
B. will transfer on the substratum that selenium content is 50PPM at the bacterial classification that growing way is stable on the 10PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
C. will transfer on the substratum that selenium content is 100PPM at the bacterial classification that growing way is stable on the 50PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
D. will transfer on the substratum that selenium content is 200PPM at the bacterial classification that growing way is stable on the 100PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
E. will transfer on the substratum that selenium content is 300PPM at the bacterial classification that growing way is stable on the 200PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
F. will transfer on the substratum that selenium content is 400PPM at the bacterial classification that growing way is stable on the 300PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
G. will transfer on the substratum that selenium content is 500PPM at the bacterial classification that growing way is stable on the 400PPM seleno culture medium and cultivate 3 days; According to the condition of a step, repeat above-mentioned steps 2 times, stable up to the bacterial classification growing way, till upgrowth situation is stable;
But h. be chosen under the selenium environment of 500PPM the still bacterial classification of normal growth.
3. preparation method according to claim 2, it is characterized in that, the final election bacterial classification finishes before transfer on the substratum that selenium content is 50PPM from 10PPM seleno culture medium domestication, at selenium content is to repeat to tame step on the substratum of 20PPM, 30PPM, 40PPM respectively once or once.
4. the application of the described S. cervisiae of claim 1 in the medicine of preparation control arseniasis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001092189A CN1142265C (en) | 2000-06-15 | 2000-06-15 | Bacterial strain resisting selenium, its preparing process and its application in preventing and treating arsenism |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001092189A CN1142265C (en) | 2000-06-15 | 2000-06-15 | Bacterial strain resisting selenium, its preparing process and its application in preventing and treating arsenism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1330139A CN1330139A (en) | 2002-01-09 |
| CN1142265C true CN1142265C (en) | 2004-03-17 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001092189A Expired - Fee Related CN1142265C (en) | 2000-06-15 | 2000-06-15 | Bacterial strain resisting selenium, its preparing process and its application in preventing and treating arsenism |
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| Country | Link |
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| CN (1) | CN1142265C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102630480B (en) * | 2012-04-01 | 2013-11-06 | 苏州安德鲁尹生物技术咨询有限公司 | Iron-rich black fungus and production method thereof |
| CN104543399B (en) * | 2015-01-20 | 2018-05-08 | 河南科技大学 | A kind of preparation method of yeast selenium forage additive |
| CN104543402B (en) * | 2015-01-20 | 2018-04-20 | 河南科技大学 | A kind of preparation method for the state iron feed additive that ferments |
| CN104543398B (en) * | 2015-01-20 | 2018-04-20 | 河南科技大学 | A kind of preparation method for the state copper feed additive that ferments |
| CN109182128A (en) * | 2018-10-16 | 2019-01-11 | 广西壮族自治区农业科学院农业资源与环境研究所 | A kind of strain culturing method of Efficient Conversion Selenium in Soil |
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2000
- 2000-06-15 CN CNB001092189A patent/CN1142265C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1330139A (en) | 2002-01-09 |
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