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RU2018143481A - MICROBIOLOGICAL METHOD FOR PRODUCING 11α-ACETOXYPROGESTERON - Google Patents

MICROBIOLOGICAL METHOD FOR PRODUCING 11α-ACETOXYPROGESTERON Download PDF

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Publication number
RU2018143481A
RU2018143481A RU2018143481A RU2018143481A RU2018143481A RU 2018143481 A RU2018143481 A RU 2018143481A RU 2018143481 A RU2018143481 A RU 2018143481A RU 2018143481 A RU2018143481 A RU 2018143481A RU 2018143481 A RU2018143481 A RU 2018143481A
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RU
Russia
Prior art keywords
extraction
microbiological method
transformation
acetoxyprogesterone
hydroxyprogesterone
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RU2018143481A
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Russian (ru)
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RU2731712C2 (en
RU2018143481A3 (en
Inventor
Татьяна Степановна Савинова
Ольга Сергеевна Савинова
Павел Николаевич Сольев
Дарья Владимировна Васина
Татьяна Владимировна Тяжелова
Татьяна Васильевна Федорова
Original Assignee
Федеральное государственное бюджетное образовательное учреждение высшего образования "Московский государственный университет имени М.В. Ломоносова" (МГУ)
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Priority to RU2018143481A priority Critical patent/RU2731712C2/en
Publication of RU2018143481A publication Critical patent/RU2018143481A/en
Publication of RU2018143481A3 publication Critical patent/RU2018143481A3/ru
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P33/00Preparation of steroids

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Botany (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Steroid Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (6)

1. Микробиологический способ получения 11α-ацетоксипрогестерона, включающий трансформацию 11α-гидроксипрогестерона штаммом мицелиального гриба Aspergillus nidulans, депонированного во Всероссийской Коллекции Промышленных Микроорганизмов под регистрационным номером F-1069.1. A microbiological method for producing 11α-acetoxyprogesterone, including the transformation of 11α-hydroxyprogesterone with a strain of mycelial fungus Aspergillus nidulans deposited in the All-Russian Collection of Industrial Microorganisms under registration number F-1069. 2. Микробиологический способ по п. 1, характеризующийся тем, что для трансформации 11α-гидроксипрогестерона штаммом мицелиального гриба Aspergillus nidulans споровую суспензию гриба высевают в качалочные колбы с ростовой средой из расчета ~1×107 спор на 100 мл среды для культивирования, выращивают при температуре 37±0,5°С в течение 4±0.5 суток до достижения количества биомассы ~1г/100 мл по весу сухого мицелия при перемешивании со скоростью 240-250 об/мин, затем в ростовую среду вносят раствор 11α-гидроксипрогестерона в биполярном апротонном растворителе до достижения концентрации растворителя в среде от 2 до 6% (об.) и до концентрации субстрата в диапазоне 0,1-1,0 г/л, и инкубируют на качалке в условиях ограниченной аэрации со скоростью 60-100 об/мин в течение 216 ч или в условиях интенсивной аэрации со скоростью 240-250 об/мин в течение не менее 168 ч; по окончании инкубации продукты извлекают экстракцией и очищают с получением целевого продукта 11α-ацетоксипрогестерона.2. The microbiological method according to claim 1, characterized in that for the transformation of 11α-hydroxyprogesterone with a strain of the mycelial fungus Aspergillus nidulans, the spore suspension of the fungus is plated in rocking flasks with growth medium at the rate of ~ 1 × 10 7 spores per 100 ml of culture medium, grown at at a temperature of 37 ± 0.5 ° C for 4 ± 0.5 days until a biomass of ~ 1g / 100 ml by weight of dry mycelium is reached with stirring at a speed of 240-250 rpm, then a solution of 11α-hydroxyprogesterone in bipolar aprotic is introduced into the growth medium solvent until the solvent concentration in the medium is from 2 to 6% (vol.) and to a substrate concentration in the range of 0.1-1.0 g / l, and incubated on a rocking chair under conditions of limited aeration at a speed of 60-100 rpm for 216 hours or in conditions of intensive aeration at a speed of 240-250 rpm for at least 168 hours; at the end of the incubation, the products are extracted by extraction and purified to obtain the desired product 11α-acetoxyprogesterone. 3. Микробиологический способ по п. 2, характеризующийся тем, что экстракцию продуктов трансформации из ферментационной суспензии проводят после предварительного разрушения клеток микроорганизма с помощью дезинтегратора-гомогенизатора.3. The microbiological method according to p. 2, characterized in that the extraction of transformation products from the fermentation suspension is carried out after preliminary destruction of the cells of the microorganism using a disintegrator-homogenizer. 4. Микробиологический способ по п. 2, характеризующийся тем, что экстракцию продуктов трансформации из ферментационной суспензии проводят методом двухэтапной экстракции - экстракции из предварительно отделенной культуральной жидкости и экстракции из мицелия мацерацией - с последующим объединением экстрактов.4. The microbiological method according to claim 2, characterized in that the extraction of the transformation products from the fermentation suspension is carried out by a two-stage extraction method — extraction from a previously separated culture liquid and extraction from mycelium by maceration — followed by combining the extracts. 5. Микробиологический способ по п. 2, характеризующийся тем, что в качестве биполярного апротонного растворителя используют диметилсульфоксид или диметилформамид.5. The microbiological method according to claim 2, characterized in that dimethyl sulfoxide or dimethylformamide is used as the bipolar aprotic solvent. 6. Микробиологический способ по п. 2, характеризующийся тем, что очистку 11α-ацетоксипрогестерона от побочного продукта трансформации - 6β, 11α-дигидроксипрогестерона - осуществляют фильтрацией через слой силикагеля в растворе дихлорметана.6. The microbiological method according to claim 2, characterized in that the purification of 11α-acetoxyprogesterone from the transformation by-product - 6β, 11α-dihydroxyprogesterone - is carried out by filtration through a layer of silica gel in a dichloromethane solution.
RU2018143481A 2018-12-07 2018-12-07 Microbiological method of producing 11α-acetoxyprogesterone RU2731712C2 (en)

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RU2731712C2 RU2731712C2 (en) 2020-09-08

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2175175T3 (en) * 1995-12-12 2002-11-16 Akzo Nobel Nv A MICROBIAL METHOD FOR THE 11ALFA-HYDROXYLATION OF STEROIDS.

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RU2018143481A3 (en) 2020-06-08

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