CN106434822A - Method for preparing high-content cholic acid by microorganism transformation method - Google Patents
Method for preparing high-content cholic acid by microorganism transformation method Download PDFInfo
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- CN106434822A CN106434822A CN201611222996.6A CN201611222996A CN106434822A CN 106434822 A CN106434822 A CN 106434822A CN 201611222996 A CN201611222996 A CN 201611222996A CN 106434822 A CN106434822 A CN 106434822A
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- cholic acid
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- fermentation
- transformation method
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
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- Engineering & Computer Science (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for preparing high-content cholic acid by microorganism transformation method, comprising steps of collecting soil sample; collecting soils at the pouring place of bile of flocks and herds and cholic acid production factory; selecting a culture, and cultivating the culture by a seed cultivating base; fermenting and cultivating; fermenting and cultivating the selected strain in a fermenting culture base; separating, purifying, crystalizing and refining, and obtaining the high-content cholic acid. The cholic acid is prepared by the microorganism transformation method; the method is featured by gentle condition, single reaction, low pollution and others. Purity of the cholic acid prepared by the method is high, and can reach 90%, and the yield is big.
Description
Technical field
The invention belongs to biological pharmacy technical field, more particularly to one kind prepare high-load gallbladder using microbe transformation method
The method of acid.
Background technology
Cholic acid is the powder of white or off-white color.Feeble QI, bitter in the mouth.It is present in Fel Bovis seu Bubali or Fel Caprae seu Ovis, is the raw material of artificial Calculus Boviss.
At present the production method of cholic acid is mainly saponification, and acidization is used the chemical reagents such as a large amount of sulphuric acid, easily causes environmental pollution,
The scientific idea for not meeting ecological, environmental protective and the production cost that increased enterprise.It is thus desirable to providing a kind of mild condition, pollution
Production method that is little, meeting environmental protection concept.
Content of the invention
The purpose of the present invention is that and overcomes above-mentioned deficiency, provide a kind of mild condition, pollute little, meet environmental protection concept
The method of production cholic acid.
For reaching above-mentioned purpose, the present invention is implemented according to technical scheme below：
A kind of method for preparing high-load cholic acid using microbe transformation method, comprises the following steps：
S1. pedotheque is gathered, gathers the soil that Fel Bovis seu Bubali or Fel Caprae seu Ovis topple over place and cholic acid production plant；
S2. strain screening, using seed culture medium culture strain；
S3. fermentation culture, the bacterial strain for screening obtained is carried out fermentation culture in the fermentation medium；
S4. separate, purify, crystallizing, refining.
Further, the seed culture medium includes following component：Potato juice 1000mL, glucose 20g, agar
200g, Fel Bovis seu Bubali or Fel Caprae seu Ovis 30g.
Further, the fermentation medium includes following component：Potato juice 1000mL, maltose 90g, agar
60g, Fel Bovis seu Bubali or Fel Caprae seu Ovis 100g.
Further, fermentative process conditions are：PH is 7.6,28 DEG C of temperature, fermentation time 48h.
Compared with prior art, beneficial effects of the present invention are：
The present invention prepares cholic acid using the method for microorganism conversion, and the method has mild condition, reacts single-minded, pollutes the spy such as little
Point.And higher by cholic acid purity obtained in the method, up to 90%, and yield is big.
Specific embodiment
With reference to specific embodiment, the invention will be further described, illustrative examples and explanation that here is invented
It is used for explaining the present invention, but not as a limitation of the invention.
Embodiment
S1. pedotheque is gathered
Collection Fel Bovis seu Bubali or Fel Caprae seu Ovis topple over the soil of place and cholic acid production plant；
S2. strain screening
（1）The pedotheque 1.0g of above-mentioned collection is taken, is put in sterilized test tube, vibrated with turbula shaker after adding sterilized water 10mL
Shake up, after standing, the supernatant is taken, drip on seed culture medium, smoothened with Glass rod, be placed in 27 DEG C of constant incubators and cultivate
2-5d, after bacterium colony is grown on flat board, chooses single bacterium colony on new seed culture medium, judges pure bacterium according to morphological characteristic
Strain, numbering is preserved；The seed culture medium includes following component：Potato juice 1000mL, glucose 20g, agar 200g, cattle
Sheep bile 30g.
（2）Tens isolated bacterial strains are seeded in seed culture medium respectively, after single bacterium colony is grown, take bacterium colony
Following culture medium 1.0g is positioned in test tube, plus 60% glacial acetic acid solution 2mL, supersound process 10 minutes, filtration, takes filtrate
1mL, puts in test tube, plus furfuryl aldehyde solution 1mL and the sulfuric acid solution of brand-new（Take sulphuric acid 50mL to mix with water 65mL）13mL, 70
Heat in DEG C water-bath, solution is as being in bluish violet, then this bacterial strain possesses conversion capability；If not showing bluish violet, do not possess conversion energy
Power；
（3）The bacterial strain that screening is obtained is cultivated on seed culture medium, and bacterium colony quality is fine and close, surface is in more velvet-like or hard
Real, dry, many wrinkle, can tentatively guess bacterial strain for actinomycetes.
（4）Picking antibacterial makes slide further, examines under a microscope, and antibacterial is thread in branch, can further determine that
Bacterial strain is actinomycetes.
S3. fermentation culture
The actinomycetes that screening is obtained are placed in fermentation medium, carry out fermentation culture, and fermentative process conditions are：PH is 7.6, temperature
28 DEG C of degree, fermentation time 48h.The fermentation medium includes following component：Potato juice 1000mL, maltose 90g, agar
60g, Fel Bovis seu Bubali or Fel Caprae seu Ovis 100g.
S4. separate, purify, crystallizing, refining
Bacterium colony and the common 1g of the culture medium below bacterium colony is taken, loaded in test tube, 95% 4 mL of ethanol is added, fully shaking is simultaneously
Heating, filters, cooling, and crystallization is filtered and obtains final product thick cholic acid, then can refine cholic acid through refined.
Technical scheme is not limited to the restriction of above-mentioned specific embodiment, and every technology according to the present invention scheme is done
The technology deformation for going out, each falls within protection scope of the present invention.
Claims (4)
1. a kind of method that high-load cholic acid is prepared using microbe transformation method, it is characterised in that comprise the following steps：
S1. pedotheque is gathered, gathers the soil that Fel Bovis seu Bubali or Fel Caprae seu Ovis topple over place and cholic acid production plant；
S2. strain screening, using seed culture medium culture strain；
S3. fermentation culture, the bacterial strain for screening obtained is carried out fermentation culture in the fermentation medium；
S4. separate, purify, crystallizing, refining.
2. the method that high-load cholic acid is prepared using microbe transformation method according to claim 1, it is characterised in that described
Seed culture medium includes following component：Potato juice 1000mL, glucose 20g, agar 200g, Fel Bovis seu Bubali or Fel Caprae seu Ovis 30g.
3. the method that high-load cholic acid is prepared using microbe transformation method according to claim 1, it is characterised in that described
Fermentation medium includes following component：Potato juice 1000mL, maltose 90g, agar 60g, Fel Bovis seu Bubali or Fel Caprae seu Ovis 100g.
4. the method that high-load cholic acid is prepared using microbe transformation method according to claim 1, it is characterised in that fermentation
Process condition is：PH is 7.6,28 DEG C of temperature, fermentation time 48h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611222996.6A CN106434822A (en) | 2016-12-27 | 2016-12-27 | Method for preparing high-content cholic acid by microorganism transformation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611222996.6A CN106434822A (en) | 2016-12-27 | 2016-12-27 | Method for preparing high-content cholic acid by microorganism transformation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106434822A true CN106434822A (en) | 2017-02-22 |
Family
ID=58215607
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611222996.6A Pending CN106434822A (en) | 2016-12-27 | 2016-12-27 | Method for preparing high-content cholic acid by microorganism transformation method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184190A (en) * | 2019-05-14 | 2019-08-30 | 成都百代禾诗生物科技有限责任公司 | Zymogenic orientation improved method for sterol fermenting and producing cholic acid |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101139380A (en) * | 2007-06-15 | 2008-03-12 | 巩利昌 | Method for producing cholic acid by using bovine and sheep bile |
| CN102676626A (en) * | 2012-05-24 | 2012-09-19 | 河南利伟生物药业股份有限公司 | Method for preparing cholesterol by microbial conversion method |
-
2016
- 2016-12-27 CN CN201611222996.6A patent/CN106434822A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101139380A (en) * | 2007-06-15 | 2008-03-12 | 巩利昌 | Method for producing cholic acid by using bovine and sheep bile |
| CN102676626A (en) * | 2012-05-24 | 2012-09-19 | 河南利伟生物药业股份有限公司 | Method for preparing cholesterol by microbial conversion method |
Non-Patent Citations (2)
| Title |
|---|
| 任凌波等: "《生物化工产品生产工艺技术及应用》", 31 December 2001 * |
| 褚庆环: "《动物性食品副产品加工技术》", 31 January 2005 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184190A (en) * | 2019-05-14 | 2019-08-30 | 成都百代禾诗生物科技有限责任公司 | Zymogenic orientation improved method for sterol fermenting and producing cholic acid |
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Application publication date: 20170222 |
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| RJ01 | Rejection of invention patent application after publication |