CN104711311A - Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process - Google Patents
Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process Download PDFInfo
- Publication number
- CN104711311A CN104711311A CN201510116569.9A CN201510116569A CN104711311A CN 104711311 A CN104711311 A CN 104711311A CN 201510116569 A CN201510116569 A CN 201510116569A CN 104711311 A CN104711311 A CN 104711311A
- Authority
- CN
- China
- Prior art keywords
- alpha
- substratum
- progesterone
- dihydroxy
- prepares
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with a one-pot continuous fermentation process, and belongs to the technical field of intermediate preparation. The method comprises the following specific steps: (1) inoculating slant culture of arthrobacter simplex into a first culture medium, performing culture for 20-30 hours, adding ground 17 alpha-hydroxyprogesterone, and performing transformation; (2) inoculating fermentation liquor after finishing the step (1) into a second culture medium, inoculating slant culture of aspergillus ochraceus into the second culture medium, performing culture, and performing post-treatment to obtain delta 1-11 alpha,17 alpha-dihydroxyprogesterone. Delta 1-11 alpha,17 alpha-dihydroxyprogesterone is prepared with the one-pot continuous fermentation process, so that a step of filtering, extracting, refining and drying a dehydrogenation material is reduced, discharge of 50% pollutants is reduced, environmental pressure is reduced, the time is saved, the yield is increased, the efficiency is improved, the process is simplified, the whole reaction operation is convenient, and the method is suitable for industrialized popularization and application.
Description
Technical field
The present invention relates to the preparing technical field of intermediate, refer to that a kind of one kettle way continuously ferments especially and prepare Δ 1-11 α, the method for 17 alpha-dihydroxy-Progesterone.
Background technology
Prednisolone acetate traditional processing technology take diene as raw material, through epoxy, walsh oxidation, fermentation hydroxylation, oxidation, upper debrominate, iodo, displacement, fermentation dehydrogenation, protection, reduction, hydrolysis, esterification totally 13 step reaction generations.
Concrete reaction formula is as follows:
Very important intermediate Δ in above-mentioned preparation process
1-11 α, 17 alpha-dihydroxy-Progesterone adopt two-step fermentation separately to carry out, every single step reaction all must by fermentation, filter, extract, refining, dry, packaging and other steps, there is yield losses in the process in process, and increase sewage discharge, consume mass energy and manpower and increase large number quipments, causing cost increase very large.It can thus be appreciated that above-mentioned complex manufacturing, complex steps, need to use some toxic reagent example hydrochloric acid Urea,amino-etc. in production process simultaneously, cause production cost high, environmental stress is large, the shortcomings such as deficiency in economic performance.
Summary of the invention
The present invention proposes a kind of one kettle way and continuously ferments and prepare Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, solves that the key intermediate complicated process of preparation of prednisolone acetate in prior art, environmental pollution are serious, the problem of yield losses.
Technical scheme of the present invention is achieved in that
One kettle way continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, comprising:
(1) slant culture of Arthrobacter simplex Atcc6942 is inoculated in the first substratum, cultivates 20-30 hour, add 17 Alpha-hydroxy Progesterone of pulverizing afterwards, transform;
(2) fermented liquid after step (1) being completed is inoculated in the second substratum; Be inoculated in the second substratum by the slant culture of Aspergillus ochraceus Aspergill usochraceus, cultivate, namely aftertreatment obtains Δ
1-11 α, 17 alpha-dihydroxy-Progesterone.
As preferred technical scheme, the proportioning following (weight percent) of described first substratum: 1% glucose, 1% corn steep liquor, 0.3% peptone, 0.15%KH
2pO
4.
As preferred technical scheme, the proportioning following (weight percent) of described second substratum: 2% glucose, 1% corn steep liquor, 1% peptone, 0.2% ammonium sulfate.
As preferred technical scheme, the fermented liquid after described step (1) completes is with in described second substratum of the inoculum size of 5%-10% access.
As preferred technical scheme, the cultivation of described step (1) carries out in shaking flask, and temperature is 25-35 DEG C.
As preferred technical scheme, the step of described step (2) aftertreatment is as follows: cross the solid mycelia cake in nutrient solution described in leaching, extracts, and heating, adds gac afterwards, then reflux and filter, and filtrate concentrates and obtains crude product; Described crude product refilters and get final product.
Technical scheme of the present invention compared with prior art, has following beneficial effect:
(1) the present invention makes total recovery compared to prior art by continuously fermenting, and total recovery reaches 81.97%, improves nearly 10 percentage point.
(2) the present invention adopts one kettle way to continuously ferment to prepare Δ 1-11 α, 17 alpha-dihydroxy-Progesterone, decrease the filtration of dehydrogen substance, extraction, refining, the step of drying, because this reducing the discharge of 50% pollutent, alleviating the pressure of environment, having saved the time, improve yield and efficiency, simplify technique, make whole operation convenient, be suitable for industrial application.
Embodiment
Be clearly and completely described to the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
The invention provides one kettle way to continuously ferment and prepare Δ 1-11 α, the method of 17 alpha-dihydroxy-Progesterone, technology of the present invention mainly makes two-step fermentation reaction directly carry out second time fermentation when not carrying out the process of middle Hydrolysis kinetics, under the acting in conjunction of two kinds of bacterium, carry out two-step reaction simultaneously.
Embodiment 1
200ml first substratum is housed in 500 ml shake flasks, and the first substratum comprises 1% (weight percent) glucose, 1% (weight percent) corn steep liquor, 0.3% (weight percent) peptone, 0.15% (weight percent) KH
2pO
4; By substratum 120 DEG C of sterilizings 30 minutes in high-pressure sterilizing pot; Be cooled to 30 ± 1 DEG C, then the slant culture of Arthrobacter simplex be inoculated in substratum; The substratum of bacterial strain is had to cultivate 24 hours in 165rpm at 30 ± 1 DEG C in shaking table inoculation.Drop into 12g through 17 Alpha-hydroxy Progesterone of comminution by gas stream, transform 48 hours.
Then add second substratum of 800ml through sterilizing with the inoculum size of 10%, the second substratum comprises 2% (weight percent) glucose, 1% (weight percent) corn steep liquor, 1% (weight percent) peptone, 0.2% (weight percent) ammonium sulfate; Then the slant culture of Aspergillus ochraceus is inoculated in substratum; The substratum of bacterial strain is had to cultivate 48 hours in 200rpm at 28 ± 1 DEG C in shaking table inoculation.
Results nutrient solution, crosses leaching solid mycelia cake, with 20 times of acetone extractions 4 times, merges and is concentrated into 20-30 times amount, combining extraction liquid, add thermosol clear after add 0.04 times of gac, backflow is filtered half an hour, filtrate is concentrated do after obtain crude product.Use chloroform-toluene=10:1, after complete molten crude product, normal pressure is concentrated into without chloroform, is cooled to 40 DEG C of filtrations, repeats the Δ that can obtain content 98% for 3 times
1-11 α, 17 alpha-dihydroxy-Progesterone fine work 8.73g, the dry after saponification of mother liquor concentrations, obtains mother liquor thing 1.35g.Total recovery 81.97%.
Embodiment 2
200ml first substratum is housed in 500 ml shake flasks, and the first substratum comprises 1% (weight percent) glucose, 1% (weight percent) corn steep liquor, 0.3% (weight percent) peptone, 0.15% (weight percent) KH
2pO
4; By substratum 120 DEG C of sterilizings 30 minutes in high-pressure sterilizing pot; Be cooled to 30 ± 1 DEG C, then the slant culture of Arthrobacter simplex be inoculated in substratum; The substratum of bacterial strain is had to cultivate 24 hours in 165rpm at 30 ± 1 DEG C in shaking table inoculation.Drop into 12g through 17 Alpha-hydroxy Progesterone of comminution by gas stream, transform 48 hours.
Then add second substratum of 800ml through sterilizing with the inoculum size of 5%, the second substratum comprises 2% (weight percent) glucose, 1% (weight percent) corn steep liquor, 1% (weight percent) peptone, 0.2% (weight percent) ammonium sulfate; Then the slant culture of Aspergillus ochraceus is inoculated in substratum; The substratum of bacterial strain is had to cultivate 48 hours in 200rpm at 28 ± 1 DEG C in shaking table inoculation.
Results nutrient solution, crosses leaching solid mycelia cake, with 20 times of acetone extractions 4 times, merges and is concentrated into 20-30 times amount, combining extraction liquid, add thermosol clear after add 0.04 times of gac, backflow is filtered half an hour, filtrate is concentrated do after obtain crude product.Use chloroform-toluene=10:1, after complete molten crude product, normal pressure is concentrated into without chloroform, is cooled to 40 DEG C of filtrations, repeats the Δ that can obtain content 98% for 3 times
1-11 α, 17 alpha-dihydroxy-Progesterone fine work 8.81g, the dry after saponification of mother liquor concentrations, obtains mother liquor thing 1.12g.Total recovery 80.97%.
Embodiment 3
200ml first substratum is housed in 500 ml shake flasks, and the first substratum comprises 1% (weight percent) glucose, 1% (weight percent) corn steep liquor, 0.3% (weight percent) peptone, 0.15% (weight percent) KH
2pO
4; By substratum 120 DEG C of sterilizings 30 minutes in high-pressure sterilizing pot; Be cooled to 30 ± 1 DEG C, then the slant culture of Arthrobacter simplex be inoculated in substratum; The substratum of bacterial strain is had to cultivate 24 hours in 165rpm at 30 ± 1 DEG C in shaking table inoculation.Drop into 12g through 17 Alpha-hydroxy Progesterone of comminution by gas stream, transform 48 hours.
Then add second substratum of 800ml through sterilizing with the inoculum size of 10%, the second substratum comprises 2% (weight percent) glucose, 1% (weight percent) corn steep liquor, 1% (weight percent) peptone, 0.2% (weight percent) ammonium sulfate; Then the slant culture of Aspergillus ochraceus is inoculated in substratum; The substratum of bacterial strain is had to cultivate 48 hours in 200rpm at 28 ± 1 DEG C in shaking table inoculation.
Results nutrient solution, crosses leaching solid mycelia cake, with 20 times of acetone extractions 4 times, merges and is concentrated into 20-30 times amount, combining extraction liquid, add thermosol clear after add 0.04 times of gac, backflow is filtered half an hour, filtrate is concentrated do after obtain crude product.Use chloroform-toluene=10:1, after complete molten crude product, normal pressure is concentrated into without chloroform, is cooled to 40 DEG C of filtrations, repeats the Δ that can obtain content 98% for 3 times
1-11 α, 17 alpha-dihydroxy-Progesterone fine work 8.76g, the dry after saponification of mother liquor concentrations, obtains mother liquor thing 1.21g.Total recovery 81.18%.
Total recovery can be increased to 81.97% by continuously fermenting by the present invention; Decrease the filtration of dehydrogen substance, extraction, refining, packaging process; Decrease the discharge of wastewater of 50%.Preparation technology of the present invention is simple, cost is low, yield high (exceeding nearly 10 percentage point than prior art), is suitable for industrial application.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (6)
1. one kettle way continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, comprising:
(1) slant culture of Arthrobacter simplex Atcc6942 is inoculated in the first substratum, cultivates 20-30 hour, add 17 Alpha-hydroxy Progesterone of pulverizing afterwards, transform;
(2) fermented liquid after step (1) being completed is inoculated in the second substratum; Be inoculated in by the slant culture of Aspergillus ochraceus in the second substratum, cultivate, namely aftertreatment obtains Δ
1-11 α, 17 alpha-dihydroxy-Progesterone.
2. one kettle way according to claim 1 continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, is characterized in that, the proportioning following (weight percent) of described first substratum: 1% glucose, 1% corn steep liquor, 0.3% peptone, 0.15%KH
2pO
4.
3. one kettle way according to claim 1 and 2 continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, is characterized in that, the proportioning following (weight percent) of described second substratum: 2% glucose, 1% corn steep liquor, 1% peptone, 0.2% ammonium sulfate.
4. one kettle way according to claim 1 continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, is characterized in that, the fermented liquid after described step (1) completes is with in described second substratum of the inoculum size of 5%-10% access.
5. one kettle way according to claim 1 continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, is characterized in that, the cultivation of described step (1) carries out in shaking flask, and temperature is 25-35 DEG C.
6. one kettle way according to claim 1 continuously ferments and prepares Δ
1-11 α, the method for 17 alpha-dihydroxy-Progesterone, is characterized in that, the step of described step (2) aftertreatment is as follows: cross the solid mycelia cake in nutrient solution described in leaching, extracts, heating, add gac afterwards, then reflux and filter, filtrate concentrates and obtains crude product; Described crude product refilters and get final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510116569.9A CN104711311A (en) | 2015-03-17 | 2015-03-17 | Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510116569.9A CN104711311A (en) | 2015-03-17 | 2015-03-17 | Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104711311A true CN104711311A (en) | 2015-06-17 |
Family
ID=53411029
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510116569.9A Pending CN104711311A (en) | 2015-03-17 | 2015-03-17 | Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104711311A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106831919A (en) * | 2015-12-04 | 2017-06-13 | 上海市农药研究所有限公司 | The Alpha-hydroxy progesterone of bioconversion 17 produces 11 α, the extraction process of 17 α-bis- hydroxyl progesterones |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102061320A (en) * | 2010-12-02 | 2011-05-18 | 浙江仙琚制药股份有限公司 | Preparation method of 11 alpha,17 alpha-dyhydroxyl-androst-4-ene-3,20-dione |
| CN102206696A (en) * | 2011-05-06 | 2011-10-05 | 浙江仙琚制药股份有限公司 | Rotor-type internal-compression oil-gas mixed transport pump unit |
| CN102827913A (en) * | 2012-08-31 | 2012-12-19 | 湖南诺凯生物医药有限公司 | Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes |
-
2015
- 2015-03-17 CN CN201510116569.9A patent/CN104711311A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102061320A (en) * | 2010-12-02 | 2011-05-18 | 浙江仙琚制药股份有限公司 | Preparation method of 11 alpha,17 alpha-dyhydroxyl-androst-4-ene-3,20-dione |
| CN102206696A (en) * | 2011-05-06 | 2011-10-05 | 浙江仙琚制药股份有限公司 | Rotor-type internal-compression oil-gas mixed transport pump unit |
| CN102827913A (en) * | 2012-08-31 | 2012-12-19 | 湖南诺凯生物医药有限公司 | Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106831919A (en) * | 2015-12-04 | 2017-06-13 | 上海市农药研究所有限公司 | The Alpha-hydroxy progesterone of bioconversion 17 produces 11 α, the extraction process of 17 α-bis- hydroxyl progesterones |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101736045B (en) | Method for continuously extracting functional components of chlorella vulgaris | |
| EP2712928B1 (en) | Method for producing natural -carotene by fermentation and use thereof | |
| CN102060934B (en) | Enzymatic extraction method for auricularia auricula polysaccharides | |
| CN100535058C (en) | Fermentation and extraction process of producing purple sweet potato haematochrome | |
| CN101255449A (en) | Use of beta-glucosidase in preparation of resveratrol by transforming polydatin | |
| CN103483040B (en) | Chinese caterpillar fungus pilot scale liquid submerged fermentation substratum and fermentation method for producing thereof | |
| CN101831471A (en) | Method for producing polysaccharides by fermenting yellow serofluid with edible and medicinal fungi | |
| CN106282274A (en) | A kind of Pichia sp. fermentation process in high density of insulin precursor protein | |
| CN111574570B (en) | Comprehensive utilization method of cordyceps militaris culture residues | |
| CN103864954B (en) | A kind of extracting method of peanut meal polysaccharides | |
| CN103070010B (en) | Coriolus versicolor strain and method for extracting Coriolus versicolor glucan by utilizing fermentation products of Coriolus versicolor strain | |
| CN103966105A (en) | Aspergillus oryzae for converting ginsenoside Rg3 to produce Rh2, production method and application | |
| CN110656148A (en) | Method for preparing dehydroepiandrosterone by converting phytosterol through resting cells | |
| CN108640956B (en) | Method for preparing flavonoid glycoside from camellia seeds | |
| CN101735331B (en) | Production process for extracting lily polysaccharides through fermentation method and product thereof | |
| CN104711311A (en) | Method for preparing delta 1-11 alpha,17 alpha-dihydroxyprogesterone with one-pot continuous fermentation process | |
| CN102643364A (en) | Method for extracting ganoderan from submerged-fermentation mycelia of ganoderma lucidum | |
| CN100529055C (en) | Bacillus alcaligenes and its application in preparing Weilan gum | |
| CN112575050A (en) | Method for preparing diosgenin by biological conversion | |
| CN109206336B (en) | Method for preparing ceramide from rice bran by fermentation method | |
| CN102703562A (en) | Method for prompting extraction of linaloe oil by utilizing flora fermentation | |
| CN106834407B (en) | Method for green production of diosgenin by biological method | |
| CN103232982B (en) | Preparation method for high-activity beta-amylase | |
| CN110628840A (en) | Method for extracting myricetin by microbial fermentation | |
| CN105462857A (en) | Marasmiellus androsaceus strain and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150617 |
|
| WD01 | Invention patent application deemed withdrawn after publication |