Method of obtaining sisomycin and micromonospora rosea s1/109 strain

专利摘要:
A process of producing sisomycin by cultivating a new species of Micromonospora.
公开号:SU1200854A3
申请号:SU802877856
申请日:1980-02-05
公开日:1985-12-23
发明作者:Гадо Иштван；Йеккель Антониа；Свобода Дьердь；Яраи Миклош；Пиукових Шандор；Иштван Шандор
申请人:Хиноин Дьедьсер Еш Ведьесети Термекек Дьяра Рт (Инопредприятие)；
IPC主号:

专利说明:


The invention relates to the microbiological industry, in particular to the production of antibiotics.
The aim of the invention is to increase the yield of the target product.
The strain Micromonospora rosea SI / 109 is characterized by the following features.
Cultural and morphological features. Macroscopic observation of a 10-day culture grown at 37 ° C on Czapek's medium shows satisfactory development, the absence of formation of aerial mycelium, the formation of convex normal round orange-yellowish-black colonies, the absence of soluble pigment.
Microscopic observation reveals long branched normal filaments without a dividing septum, 0.5 M in diameter, spores round to the egg-shaped ones located in the simple sporangus x
1-1.5f.
Peptone - iron-o-agar. Growth is weak, the color is warm yellow, slightly gray; small spore layer; soluble pigment is missing.
Agars Emerson. Growth is weak. 12 colonies, orange, light, slightly brown.
Glucose - orange agar extract. Growth is satisfactory, the color is bright orange, slightly black, the colonies are highly wrinkled.
Nutrient agar. Growth is satisfactory, the color of the colonies is bright orange, slightly black.
Agar čapek. Growth is satisfactory, the color is bright-org1nzhevy, then blacks.
Agar Bennett. Growth is satisfactory, the color is slightly orange, slightly brown, forms a soluble pigment.
Sliced potatoes (with and without calcium carbonate). Growth in the tracks, the color is orange.
Physical and biochemical characteristics. It grows well at 28-37 C and does not develop at 44 C. Carbohydrate absorption (tested on medium containing 0.5% yeast extract, 1% carbon source, 0.1% CaCO, 1.5% agar, and distilled water). Absorbs xylose, fructose, gliekosis.
008542
sucrose, starch, ribose; assimilates worse galactic goa, rafnnuzu, dultsit; poorly absorbs arabinose, rhamnosis, lactose, mannitol, inositol.
5 Assimilate nitrogen (determination on medium containing 1% glucose, 1.5% agar, distilled water).
Well absorbed 0, yeast extract, 1% N-Z-amine, mediocre - 1% asparagine, poorly -% glutamic acid and 1% ammonium nitrate.
Gelatin, milk, and starch hydra15. Nitrate to nitrite restores. Maximum salt tolerance for sodium chloride is 2%.
Example i. 1 ml containing 10 -10 cells of strain 5 1/109,
20 stored at -20 ° C, boil in a series of 3000 ml flasks, each of which contains 800 ml of steroid Hoi media containing Tryptone (Oxoid) 5 g, soy flour 10 g,
5 soluble starch 20 g, glucose I g, calcium carbonate 2 g, tap WATER up to 1000 ml.
Cultivation was carried out at 28 ° C for 3 days with constant stirring at a speed of 260 rpm.
/ 1l of preparation of the inoculum obtained in the previous stage, the culture is subcultured in 100 liters of sterile medium of the following composition: soybean
5 flour 10 g, pancreatic casein hydrolyzate (dry weight) 5 g, potato starch 22 g, glucose 10 g, calcium carbonate 2 g, palm oil 2 g, water
0 water to 1000 ml of pH before sterilization 7.0.
The cultivation is carried out for 36-48 hours at 30 ° C and constant stirring at a speed of 400 rpm.
5 and aeration rates of 1/1 V / Y. Palm oil is added as a defoaming agent.
100 l of fermentation medium of the following composition, g: soy flour
0 40, peptone 10, potato starch 10, dextrin 25, glucose 5, calcium carbonate 4, yagni sulphate heptahydrate 2, iron ip cep ioacid heptahydrate 0.2, cobalt nitric acid 0,0008, palm oil 2, tap water up to 1, and inoculated 10 l inoculum torus. Fermentation is carried out under stirring at a speed of 400 rpm for 1 hour at 121 ° C, a pressure of 1.31, 4 atm, and aeration rate of 1/1 V / Y of the fermentation medium prior to sterilization 8.0; fermentorschu lead in the fermenter stainless steel.
The production of the antibiotic starts from 35-40 hours of fermentation and reaches a maximum of 110-120 hours. The end time of the fermentation is established in a turbulimetric manner.
At the end of the fermentation, the total activity of the culture liquid is 650 U / ml, based on the standard of sisomidin flE, which corresponds to the activity of 1 µg of the standard of the sisomicin base relative to epidermidis staphylococcus; determination was carried out by the method of diffusion into agar).
EXAMPLE 2 Obtained in Example 1 in an amount of 100 liters was added to a 1 m fermentation medium of the following composition, g: soybean meal 40, pancreatic casein hydrolyzate (dry weight) 10, corn starch 50, glucose 5 , angle 008544
acidic calcium 4, magnesium sulphate heptahydrate 2, iron sulphate heptagindrate 0.2, cobalt nitrate hexahydrate 0.0008, palm oil 2, tap water up to I l.
pH of the medium before steri: tization 8.0, Sterilization of the medium, mixing and aeration of the culture fluid are carried out according to example 1.
The total activity of the culture liquid is 680 U / ml. 15 g of the isolated pure base are dissolved in 60 ml of distilled
15 water and using 5 n, a solution of sulfuric acid to establish a pH of 4.3. The solution is mixed with 1.5 g of activated carbon for 30 minutes and filtered through a Zeitua filter.
The filter is washed three times with 5 ml of deionized water and the combined filtrates are poured into 1 L of methanol with stirring. After cooling, the precipitate is filtered off, the residue is washed three times with 50 ml of methanol and dried under vacuum at 50 ° C. 22 g of sisomycin sulfate are obtained.

权利要求:
Claims (3)
[1]
1. A method of producing sisomycin by cultivating a producer strain of the Micromonospora species under aerobic conditions in a nutrient medium containing carbon and nitrogen sources and mineral salts, followed by isolation of the target product in the form of a base from the culture fluid, if necessary, purifying it and transferring it to a salt that differs the fact that, in order to increase the yield of the target product, the strain Micromonospora rosea SI / 109 MNG 00182 is used as a producer and the cultivation is carried out in a fermentation culture medium,
containing, g / l of water: Soya flour 40 Peptone or hydro casein lysate 10 Potato or corn starch 10-50 Glucose 5 Calcium carbonate four Sulfuric acid heptahydrate magnesium logo 2 Sulfate Teptahydrate iron logo 0.2 Nitrogen hexahydrate cobalt acid 0,0008 Palm oil 2. 2. The method according to claim 1, about t l and h a N and i that nourishing
the medium additionally contains dextrin or soluble starch in an amount of 20-25 g / l of water.
[2]
3. The method of pop. 1, characterized in that sysomycin - base is converted into sulphate salt of sisomycin.
[3]
4.Strain Micromonospora rosea
SI / 109, MNG 00182 (Hungarian National Collection, Budapest) - producer of sisomycin.
I 200854

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引用文献:

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US3832286A|1971-02-03|1974-08-27|Schering Corp|Sisomicin and methods for its production|

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US3956068A|1972-07-14|1976-05-11|Schering Corporation|Antibiotic G-52 and method for the production thereof|US4977084A|1989-02-16|1990-12-11|Board Of Trustees Operating Michigan State University|Production, isolation, and identification of novel antifungal compounds|

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US5303518A|1993-02-11|1994-04-19|Strickland Industries, Inc.|Lined manhole assembly and liner|

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US7794713B2|2004-04-07|2010-09-14|Lpath, Inc.|Compositions and methods for the treatment and prevention of hyperproliferative diseases|

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US7862812B2|2006-05-31|2011-01-04|Lpath, Inc.|Methods for decreasing immune response and treating immune conditions|

法律状态:

优先权:

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申请号 | 申请日 | 专利标题

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HU79CI1911A|HU179146B|1979-02-06|1979-02-06|Process for microbiological producing sysnycin with fermenting micromonospora rosea|

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