CN103374537A - Method for preparing enduracidin and strain produced thereby - Google Patents
Method for preparing enduracidin and strain produced thereby Download PDFInfo
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- CN103374537A CN103374537A CN2012101243980A CN201210124398A CN103374537A CN 103374537 A CN103374537 A CN 103374537A CN 2012101243980 A CN2012101243980 A CN 2012101243980A CN 201210124398 A CN201210124398 A CN 201210124398A CN 103374537 A CN103374537 A CN 103374537A
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- enramycin
- fermentation
- enduracidin
- strain
- fermentation culture
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Abstract
The invention relates to a method for preparing enduracidin and a strain produced thereby. The strain disclosed by the invention is streptomyces fungicidius (Streptomyces fungicidius) BJX002; the preservation number is CGMCC No.6026. The method comprises the following steps: 1) inoculating seed solution of the strain into fermentation medium to ferment and cultivate, so as to obtain a fermentation liquor; 2) extracting the enduracidin from the fermentation liquor. The strain disclosed by the invention has strong ability of producing effective components A and B of the enduracidin; 10,000 mug/ml of fermentation liquor can be achieved; furthermore, the ability of producing the effective components A and B of the enduracidin is still kept at the original level after the strain disclosed by the invention is continuously inherited five generations; the genetic stability is good. By adopting the method for preparing the enduracidin by the strain disclosed by the invention, the efficiency of producing the enduracidin can be improved; the method is simple, low in cost and suitable for popularization and application in production of the enduracidin.
Description
Technical field
The present invention relates to a kind of method and production bacterial strain thereof for preparing enramycin.
Background technology
Enramycin (Enramycin) has another name called Enramycin, enramycin, enramycin, enramycin.It is a kind of polypeptide antibiotics that is combined into by unsaturated fatty acids and 17 seed amino acids that is produced by the actinomycetes of separating in the soil (Streptomyces fungicidius No.B5477) fermentation, difference according to terminal fatty acid molecule is divided into enramycin A and enramycin B with it, and enramycin is the mixture of these two kinds of components.This medicine was researched and developed by Japanese Takede Chemical Industries Ltd in 1966, was registered and was widely used in many countries thereafter at Japanese official registration in 1974.1993, China Ministry of Agriculture ratified this medicine and registers in China.2005, domestic production enterprise and U.S. Schering Plough animal health company limited (Schering Plough Anima Health Corp.) beginning joint production enramycin premix.
Enramycin is polypeptide antibiotics, and its Antibacterial mechanism is the synthetic of anti-bacteria cell walls.Enramycin has very strong activity to gram positive bacterium, particularly the harmful clostridium in the enteron aisle is had very strong inhibitory or killing effect, is 0.39～3.12g/ml to the minimum inhibition concentration (MIC) of gram positive bacterium.Enramycin can also change by the Growth and reproduction that suppresses unwanted bacteria in the enteron aisle colony balance in the enteron aisle, and the absorption of nutritive substance and utilization increase in the feed.The life-time service enramycin, bacterium also is difficult for producing resistance to it, does not also produce obvious crossing drug resistant between enramycin and other microbiotic.Because enramycin possesses above plurality of advantages, makes it have good social benefit and environmental benefit, is a kind of novel green antibiotic, has wide market outlook.
And in fermentation industry, improve the production efficiency of unit bacterial classification, can under the existing installation condition, improve output on the basis that does not increase raw materials cost, power cost, human cost.
Summary of the invention
The purpose of this invention is to provide a strain kabicidin streptomycete to improve the efficient of producing enramycin.
Another purpose of the present invention provides the method that bacterial strain of the present invention is produced enramycin of using.
Kabicidin streptomycete provided by the present invention is kabicidin streptomycete (Streptomyces fungicidius) BJX002, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 23rd, 2012 and (be called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), Classification And Nomenclature is kabicidin streptomycete (Streptomyces fungicidius), and preserving number is: CGMCC No.6026.
The fibrillae of spores of kabicidin streptomycete BJX002 twist, the spore sphere is to oval, intermediate projections, smooth surface, aerial hyphae is canescence on synthetic medium, substrate mycelium is khaki color.The gas silk is white to grey on glycerine nitrate nutrient agar, and the base silk is colourless.The gas silk is extremely little grey of white on glucose asparagine nutrient agar, and the base silk is colourless, but lysochrome is little yellow sometimes.The gas silk is extremely little grey of white on the Starch Agar substratum, and the base silk is colourless to little yellow.The gas silk is extremely little grey of white on the calcium malate nutrient agar, and the base silk is colourless to little yellow.But lysochrome without, some strain is pink colour, rear disappearance.On glycerine asparagine agar (ISP), inorganic salt Starch Agar (ISP), yeastex malt extract agar (ISP), oatmeal agar (ISP) substratum the gas silk without or few, be white.Base silk reverse side is without differentiating pigment: light yellow or light lark.But without lysochrome.But on nutrient agar lysochrome without or tawny slightly.On the potato ball substratum gas silk without or white to little grey, the little yellow of base silk may have the Vandyke brown element to little grey around the base silk.This bacterial strain can make gelatine liquefication, and a little less than milk solidified and peptonizes, hydrolyzed starch was strong.Nitrate does not reduce or is suspicious.Do not produce Melanoidins, tyrosine oxidase and H
2S.Utilize D-Glucose, D-wood sugar, D-Fructose, inositol, PEARLITOL 25C; Utilize suspicious to L-arabinose, sucrose, rhamnosyl, raffinose.
The invention provides a kind of method for preparing enramycin, comprise the steps:
1) fermentation culture: the seed liquor of described bacterial strain is inoculated in fermentation culture obtains fermented liquid in the fermention medium;
2) from fermented liquid, extract enramycin.
Wherein, described strain culturing is to cultivate 192-240 hour under temperature 25-30 ℃ condition.
Described fermentation culture based component is: W-Gum 7.0-13.0g/L, glucose 9.1-16.9g/L, dipotassium hydrogen phosphate 0.14-0.26g/L, magnesium sulfate heptahydrate 0.7-1.3g/L, sodium-chlor 10.5-19.5g/L, calcium carbonate 1.54-2.86g/L.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the described fermention medium sterilization.
Described fermention medium is preferably as follows substratum: W-Gum 10g/L, glucose 13g/L, dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate heptahydrate 1g/L, sodium-chlor 15.2g/L, calcium carbonate 2.2g/L.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the described fermention medium sterilization.
Inoculum size is preferably 5～15% (V/V), and described seed liquor is the seed liquor that is cultured to logarithmic phase.
Described fermentation can be carried out under the condition of vibration, and the rotating speed of described vibration can be 170-210rpm, and rotation radius is 50mm.The rotating speed of described vibration is preferably 190rpm.
In order to prevent that fermented liquid from concentrating because too much evaporating, cause false fermentation unit, ambient relative humidity is 50-60% described in the maintenance fermenting process.
The temperature of described fermentation is preferably 28 ℃.
The time of described fermentation is preferably 240 hours.
The ability that bacterial strain of the present invention is produced enramycin is strong, can reach 10000 μ g/ml fermented liquids, and active princlple A and B ratio reach more than 3.0.And bacterial strain of the present invention passed continuously for 5 generations after its ability and A/B ratio of producing enramycin all also remain on previous level, show that its genetic stability is good.Prepare the method for enramycin with bacterial strain of the present invention, can improve the efficient of producing enramycin, and method is simple, with low cost.Therefore, bacterial strain of the present invention and the method for preparing enramycin are suitable for applying in the production of enramycin.
Description of drawings
Fig. 1 is the high-efficient liquid phase chromatogram of enramycin standard substance.
Fig. 2 is the high-efficient liquid phase chromatogram of enramycin in the ferment filtrate.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
W-Gum is available from Heilongjiang Longfeng Corn Development Co., Ltd.; Glucose is available from Jingpo Lake, Heilongjiang Province agricultural development company limited, the corn starch is available from Zouping, Shandong crane concomitant product Science and Technology Ltd., peptone is available from Yanshi City various schools of thinkers industry and trade company limited, sodium-chlor is available from imperial salt (Mudanjiang) salt industry company limited, calcium carbonate is already company limited of imperial calcium available from Hebei province's hole diameter, dipotassium hydrogen phosphate and magnesium sulfate heptahydrate are available from the huge chemicals of Tianjin Dongli District factory, agar is available from Qingdao whale marine life company limited, yeast extract paste is available from biotechnology company limited of Wuxue City state nation, and asparagine is available from chemical reagent Beijing company limited of traditional Chinese medicines group.
The preparation of embodiment 1 bacterial strain
One, the preparation of bacterial strain
The high-throughput prescreening method of using in this experiment is as follows:
Fermentation stage: adopt 96 hole depth orifice plates, dress substratum 0.3ml in every hole.Every hole access bacterial classification, 28 ℃ leave standstill cultivation 10 days.Used medium is by W-Gum 10g/L, glucose 13g/L, and dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate heptahydrate 1g/L, sodium-chlor 15.2g/L, calcium carbonate 2.2g/L and agar 2.5g/L form, and solvent is water.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the sterilization.
The extraction stage: every hole is soaked with 1ml methyl alcohol, smash to pieces, get supernatant.
The mensuration stage: measure the UV absorption value of supernatant under 267nm with microplate reader.
1, isolated strains from soil
Get the soil on limit, Jingpo Lake, Ningan City, Heilongjiang Province, soil is made suspension, Soil Slurry is inoculated in the isolation medium cultivates, picking list bacterium colony dilutes the line separation and purification, obtains the pure culture bacterial strain; Isolation medium is by glucose 20g/L (adding after the sterilization separately), agar 20g/L, yeast extract paste 2g/L, asparagine 0.5g/L and K
2HPO
40.5g/L form, solvent is water; PH is 7.2; 115 ℃, sterilized 20 minutes;
Primary dcreening operation: the pure culture bacterial strain of acquisition carries out shake flask fermentation to be cultivated, fermented liquid centrifugal 5 minutes through 8000rpm, and supernatant liquor obtains ferment filtrate through the filtering with microporous membrane of 0.45 μ m; The HPLC analytical method detects in the fermented liquid whether contain enramycin; The bacterial strain that can produce enramycin carries out respectively following serial mutagenesis.
2, mutagenesis
(1) ultraviolet compounded Streptomycin sulphate mutagenesis
Get bacteria suspension 7ml, add in the sterile petri dish, shine in the 30cm place with the uv irradiating instrument of 30W.Set irradiation time and be respectively 60s, 90s.After the isolation medium sterilization, add the after filtration Streptomycin sulphate of degerming of final concentration 0.1 μ g/ml, 0.2 μ g/ml, paved plate is cultivated 10d for 28 ℃.Picking list bacterium colony carries out high flux screening, shaking flask retrial according to the method described above.The superior strain that obtains carries out next step mutagenesis.
(2) ultraviolet compounded Streptomycin sulphate mutagenesis (the 2nd takes turns)
The mutagenic obtained superior strain of ultraviolet compounded Streptomycin sulphate of the first round used the same method again carry out ultraviolet compounded Streptomycin sulphate mutagenesis.Cultivate 10d for 28 ℃.Picking list bacterium colony carries out high-throughput primary dcreening operation, shaking flask retrial.The superior strain that obtains carries out next step mutagenesis.
(3) 5 FU 5 fluorouracil
With the bacterial strain inclined plane inoculating in the hungry substratum without organic nitrogen source, overnight incubation, in the 5 FU 5 fluorouracil plate that contains different concns, the final concentration of 5 FU 5 fluorouracil is respectively in the 5 FU 5 fluorouracil plate of 50 μ g/ml, 100 μ g/ml with inoculation.Cultivate 10d for 28 ℃.Picking list bacterium colony carries out high-throughput primary dcreening operation, shaking flask retrial.The superior strain that obtains carries out next step mutagenesis.
(4) microwave combined streptomycin mutagenesis
650W household microwave oven take pulse-repetition as 2450MHz carries out radiotreatment to bacteria suspension in the plate respectively.Make the plate cooling behind each irradiation 5s, shine again, and with irradiation time accumulative total, the accumulative total irradiation time is respectively 60s, 90s.After the isolation medium sterilization, add the after filtration Streptomycin sulphate of degerming of final concentration 0.1 μ g/ml, 0.2 μ g/ml, paved plate.Cultivate 10d for 28 ℃.Picking list bacterium colony carries out high-throughput primary dcreening operation, shaking flask retrial.The superior strain that obtains carries out next step mutagenesis.
(5) NTG (nitrosoguanidine) combined streptomycin mutagenesis
Nitrosoguanidine (NTG) mutagenesis 0.1mol/L, the phosphate buffered saline buffer of pH6.0 prepares spore suspension, it is respectively the NTG processing monospore suspension of 1mg/ml, 2mg/ml and 4mg/ml concentration with final concentration, in 28 ℃ of concussion 30min, with physiological saline centrifuge washing spore 3 times, after dilution, coat and contain the after filtration plate of the Streptomycin sulphate of degerming of final concentration 0.1 μ g/ml, 0.2 μ g/ml.Cultivate 10d for 28 ℃.Picking list bacterium colony carries out high-throughput primary dcreening operation, shaking flask retrial.From screening, obtain the superior strain of enramycin.
Through above-mentioned series methods, finally obtained enramycin superior strain BJX002.
Two, the evaluation of bacterial strain
Kabicidin streptomycete bacterial strain BJX002 is aerobic actinomycetes, fibrillae of spores twist, the spore sphere is to oval, intermediate projections, smooth surface, aerial hyphae is canescence on synthetic medium, substrate mycelium is khaki color.
By the 16SrDNA of this bacterium of pcr amplification, the used primer sequence of PCR is:
Forward primer: 5 ' AGAGTTTGATCCTGGCTCAG3 '
Reverse primer: 5 ' AAGTCGTAACAAGGTAGCCGTA3 '
The PCR product is carried out gel electrophoresis, find identically with the purpose stripe size, be 1481bp.Sequence is shown in sequence table 1.
Above physiological and biochemical property shows that the mutant strain that the present invention obtains belongs to kabicidin streptomycete (Streptomyces fungicidius), with its called after BJX002, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 23rd, 2012 and (is called for short CGMCC, address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC No.6026.
Embodiment 2 fermentation kabicidin streptomycin B JX002 produce enramycin
The application responds Surface Method is optimized superior strain BJX002 fermentation culture conditions.
Slant medium forms: by W-Gum 10g/L, and glucose 13g/L, corn starch 10g/L, dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate heptahydrate 1g/L, sodium-chlor 15g/L, calcium carbonate 2.2g/L forms, and solvent is water.The pH value of slant medium is 7.2,121 ℃, sterilization 30min.
Seed culture medium forms: by W-Gum 20g/L, and glucose 25g/L, corn starch 10g/L, peptone 5g/L, dipotassium hydrogen phosphate 0.1g/L, sodium-chlor 10g/L, calcium carbonate 5g/L forms, and solvent is water.The pH value of seed culture medium is 7.2,121 ℃, sterilization 30min.
Fermention medium I forms: by W-Gum 7.0g/L, and glucose 9.1g/L, dipotassium hydrogen phosphate 0.14g/L, magnesium sulfate heptahydrate 0.7g/L, sodium-chlor 10.5g/L, calcium carbonate 1.54g/L forms, and solvent is water.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the sterilization.
Fermention medium II forms: by W-Gum 13.0g/L, and glucose 16.9g/L, dipotassium hydrogen phosphate 0.26g/L, magnesium sulfate heptahydrate 1.3g/L, sodium-chlor 19.5g/L, calcium carbonate 2.86g/L forms, and solvent is water.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the sterilization.
Fermention medium III forms: by W-Gum 10g/L, and glucose 13g/L, dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate heptahydrate 1g/L, sodium-chlor 15.2g/L, calcium carbonate 2.2g/L forms, and solvent is water.Regulate pH to 7.2 with the 100g/L sodium hydroxide solution before the sterilization.
One, utilize fermention medium I fermentation kabicidin streptomycete BJX002 to prepare enramycin
(1) fermentation kabicidin streptomycete BJX002
1, slant culture:
Kabicidin streptomycete BJX002 is inoculated on the slant medium, is to cultivate 10-12 days under the 50-60% condition at 28 ℃, ambient relative humidity.
2, seed culture
Take the inclined-plane lawn 1cm that obtains in the step 1
2The went out seed bottle of seed culture medium of bacterium of 30ml is equipped with in its access, cultivate under the following conditions 44-48 hour (to logarithmic phase): temperature is that 28 ℃, ambient relative humidity are that 50-60%, rotating speed are that 180-200rpm, rotation radius are 50mm, obtains seed culture fluid.
3, fermentation culture
Fermentation culture method: get above-mentioned seed culture fluid and be inoculated in by the inoculum size of 10% (V/V) and 30ml is housed went out in the triangular flask of fermention medium I of bacterium, fermentation culture is 240 hours under the following conditions: temperature is that 28 ℃, humidity are 55%, rotating speed is that 200rpm, rotation radius are 50mm, obtains fermented liquid.
(2) extracting enramycin and output detects
1, extracting method is as follows:
(1) get the 2ml fermented liquid, adding fills in the 10ml ground colorimetric cylinder of 8ml methyl alcohol, and ultrasonic echography 20min with the centrifugal 10min of the speed of 4000rpm, gets supernatant solution, obtains altogether the 9.2-9.4ml supernatant.
(2) get the 1ml supernatant, with the organic phase filtering with microporous membrane of 0.45 μ m, obtain the fermentation vat liquor, to be measured.
2, HPLC analyzes:
Adopt Shimadzu LC-10A type high performance liquid chromatograph, the stainless steel column of 250mm * 4.6mm (id), in fill out the C18 reversed-phase column that Reliasil C18 model is Wondasil, 35 ℃ of column temperatures are got the fermentation vat liquor of step (2), sample size 20 μ l; Take phosphate-buffered salt (pH8.2): acetonitrile (volume ratio was as 7: 3) separates as moving phase, and flow velocity is 1ml/min, utilizes the UV detector to detect at wavelength 267nm place and automatically forms separating spectrum; Under this chromatographic condition, the color atlas of enramycin standard substance as shown in Figure 1, the retention time of enramycin active princlple A and B is respectively about 16.5min and 28.5min.
Retention time is the elution peak area of locating about 16.5min and 28.5min in the calculating ferment filtrate, calculates the content of enramycin A and B.The color atlas of enramycin as shown in Figure 2 in the ferment filtrate.
3 repetitions are established in experiment, and the result takes the mean.The result shows, obtains the A component of 162.6 μ g/ml and the B component of 34.6 μ g/ml, A: B=4.7 in the 20 μ l ferment filtrates.Adjust to such an extent that enramycin output is 9858 μ g/ml.
Two, utilize fermention medium II fermentation kabicidin streptomycete BJX002 to prepare enramycin
(1) fermentation kabicidin streptomycete BJX002
1, slant culture:
With consistent described in the step 1.
2, seed culture:
With consistent described in the step 1.
3, fermentation culture:
Fermentation culture method: get above-mentioned seed culture fluid and be inoculated in by the inoculum size of 7% (V/V) and 30ml is housed went out in the bottle of fermention medium II of bacterium, fermentation culture is 192 hours under the following conditions: temperature is that 26 ℃, ambient relative humidity are 50%, rotating speed is that 200rpm, rotation radius are 50mm, obtains fermented liquid.
(2) extracting enramycin and output detects
1, extracting method is as follows:
With consistent described in the step 1.Obtain the 9.4-9.5ml supernatant with the 2ml broth extraction.Get the 2ml supernatant, with the filtering with microporous membrane of 0.45 μ m, obtain the fermentation vat liquor, to be measured.
2, HPLC analyzes
Method is with consistent described in the step 1.
3 repetitions are established in experiment, and the result takes the mean.The result shows, obtains the A component of 156.7 μ g/ml and the B component of 34.8 μ g/ml, A: B=4.5 in the 20 μ l ferment filtrates.Adjust to such an extent that enramycin output is 9576 μ g/ml.
Three, utilize fermention medium III fermentation kabicidin streptomycete BJX002 to prepare enramycin
(1) fermentation kabicidin streptomycete BJX002
1, slant culture:
With consistent described in the step 1.
2, seed culture:
With consistent described in the step 1.
3, fermentation culture:
Fermentation culture method: get above-mentioned seed culture fluid and be inoculated in by the inoculum size of 10% (V/V) and 30ml is housed went out in the bottle of fermention medium III of bacterium, fermentation culture is 216 hours under the following conditions: temperature is that 30 ℃, ambient relative humidity are 60%, rotating speed is that 190rpm, rotation radius are 50mm, obtains fermented liquid.
(2) extracting enramycin and output detects
1, extracting method is as follows:
With consistent described in the step 1.Obtain the 9.2-9.4ml supernatant with the 2ml broth extraction.Get the 1ml supernatant, with the filtering with microporous membrane of 0.45 μ m, obtain the fermentation vat liquor.
2, HPLC analyzes
Method is with consistent described in the step 1.
3 repetitions are established in experiment, and the result takes the mean.The result shows, obtains the A component of 166.8 μ g/ml and the B component of 36.3 μ g/ml, A: B=4.6 in the 20 μ l ferment filtrates.Adjust to such an extent that enramycin output is 10156 μ g/ml.
Embodiment 3 bacterial strain genetic stabilities of the present invention detect
Mutant strain BJX002 is carried out the inclined-plane to go down to posterity, passed altogether for 5 generations, in per generation,, the cultural method of bacterium was all consistent as described in example 2 above, and the bacterium that every culture obtains is all fermented according to method described in the step 1 among the embodiment 2 and extracts, and calculated the ability of the enramycin that bacterial strain produces.
3 repetitions are established in experiment, and the result takes the mean.Bacterial strain BJX002 of the present invention is after going down to posterity for 5 times as a result, and throughput can also keep original level, shows that the genetic stability of bacterial strain BJX002 of the present invention is good.
Claims (10)
1. kabicidin streptomycete (Streptomyces fungicidius) BJX002, its preserving number is CGMCC No.6026.
2. a method that adopts the described bacterial strain of claim 1 to prepare enramycin is characterized in that, may further comprise the steps:
1) fermentation culture: the seed liquor of described bacterial strain is inoculated in fermentation culture obtains fermented liquid in the fermention medium;
2) from fermented liquid, extract enramycin.
3. method according to claim 2 is characterized in that, described step 1) fermentation culture be under temperature 25-30 ℃ condition, to cultivate 192-240 hour.
4. according to claim 2 or 3 described methods, it is characterized in that, described fermentation culture based component is: W-Gum 7.0-13.0g/L, glucose 9.1-16.9g/L, dipotassium hydrogen phosphate 0.14-0.26g/L, magnesium sulfate heptahydrate 0.7-1.3g/L, sodium-chlor 10.5-19.5g/L, calcium carbonate 1.54-2.86g/L regulates pH to 7.2 before the described fermention medium sterilization.
5. method according to claim 4, it is characterized in that, described fermentation culture based component is: W-Gum 10g/L, glucose 13g/L, dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate heptahydrate 1g/L, sodium-chlor 15.2g/L, calcium carbonate 2.2g/L regulates pH to 7.2 with the 100g/L sodium hydroxide solution before the described fermention medium sterilization.
6. according to claim 2 or 3 described methods, it is characterized in that described fermentation is carried out under the condition of vibration, the rotating speed of described vibration is 170-210rpm, and rotation radius is 50mm.
7. method according to claim 6 is characterized in that, the rotating speed of described vibration is 190rpm.
8. according to claim 2 or 3 described methods, it is characterized in that the ambient relative humidity of described fermentation is 50-60%.
9. method according to claim 3 is characterized in that, described temperature is 28 ℃, and incubation time is 240 hours.
10. the application of the described kabicidin streptomycete of claim 1 in producing enramycin.
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| CN106148460A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | A kind of fermentation medium improving enramycin B component yield and fermentation process |
| CN110072881A (en) * | 2016-12-06 | 2019-07-30 | 英特维特国际股份有限公司 | Cabicidin streptomycete isolate of modification and application thereof |
| CN110876797A (en) * | 2019-12-02 | 2020-03-13 | 广东容大生物股份有限公司 | Method for preparing enramycin dry suspension |
| US11447530B2 (en) | 2016-12-06 | 2022-09-20 | Oregon State University | Compositions and methods for enhanced production of enduracidin in a genetically engineered strain of streptomyces fungicidicus |
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| CN103740612A (en) * | 2013-12-19 | 2014-04-23 | 河北圣雪大成制药有限责任公司 | High-yield enramycin strain and screening method thereof |
| CN106148460A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | A kind of fermentation medium improving enramycin B component yield and fermentation process |
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| JP2020500509A (en) * | 2016-12-06 | 2020-01-16 | インターベット インターナショナル ベー． フェー． | Modified Streptomyces fungisidicus isolates and uses thereof |
| US11447530B2 (en) | 2016-12-06 | 2022-09-20 | Oregon State University | Compositions and methods for enhanced production of enduracidin in a genetically engineered strain of streptomyces fungicidicus |
| JP2023012549A (en) * | 2016-12-06 | 2023-01-25 | インターベット インターナショナル ベー． フェー． | Modified streptomyces fungicidicus isolates and use thereof |
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