CN106434444A - Streptococcus equi subsp and production technology for preparing hyaluronic acid through streptococcus equi subsp - Google Patents
Streptococcus equi subsp and production technology for preparing hyaluronic acid through streptococcus equi subsp Download PDFInfo
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- CN106434444A CN106434444A CN201610844980.2A CN201610844980A CN106434444A CN 106434444 A CN106434444 A CN 106434444A CN 201610844980 A CN201610844980 A CN 201610844980A CN 106434444 A CN106434444 A CN 106434444A
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- hyaluronic acid
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/46—Streptococcus ; Enterococcus; Lactococcus
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- C12N1/00—Microorganisms, 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
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Abstract
The invention discloses a streptococcus equi subsp and a production technology for preparing hyaluronic acid through the streptococcus equi subsp. According to the novel screened streptococcus equi subsp, the improved fermentation technology is combined, the output rate of a hyaluronic acid product is high, and can be 9.5 g/L. The fermented streptococcus equi subsp is easy to separate and purify, and the production cycle is short; production is easy to control, the streptococcus equi subsp is suitable for industrial mass production, the yield is high, and the production cost is greatly reduced.
Description
Technical field
The present invention relates to hyaluronic acid production technical field, more particularly to a kind of Malian drainage and made with which
The production technology of standby hyaluronic acid.
Background technology
The technique fermentation period that Production by Microorganism Fermentation hyaluronic acid is adopted at present is long, and fermentation technology can not be according to strain
Characteristic is carried out, and causes bacterial activity be excited, and hyaluronic acid productivity is low, poor product quality.The new fermentation strain of exploitation,
Meanwhile, improve production technology to improve quality and the yield of hyaluronic acid according to strain properties, be the skill of present invention needs solution
Art problem.
Content of the invention
The purpose of the present invention is aiming at above-mentioned defect and provides a kind of Malian drainage and its preparation
The production technology of hyaluronic acid.The present invention using the new Malian drainage for filtering out, in conjunction with the fermentation skill after improvement
Art, hyaluronic acid product output capacity height, up to 9.5g/L.The hyaluronic acid that fermentation is obtained is easily isolated purification, with short production cycle.
Easily controllable, suitable industrialization large-scale production, high income is produced, and greatly reduces production cost.
A kind of Malian drainage of the present invention and its prepare the production Technology scheme of hyaluronic acid and be, a kind of
Malian drainage（Streptococcus equi subsp. zooepidemicus）DCS005, the horse hammer at present
Bacterium epizootic disease subspecies are in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, address：Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, postcode：100101, its deposit number is：CGMCC No.12782, the Latin title of strain isStreptococcus equi subsp. zooepidemicus, join the microorganism of evidence（Strain）：DCS005, preservation date is
On July 14th, 2016.
The production technology of hyaluronic acid is prepared with described Malian drainage, is comprised the following steps：
（1）Shake-flask seed liquid culture；
（2）Strain expanded culture；
（3）Fermentation, is first passed through the fermentation of aseptic oxygen-containing air, then is passed through nitrogen continuation fermentation；
（4）Purification.
Step（1）、（2）、（3）In culture fluid include starch 1% by weight percentage, sucrose 0.5%, Carnis Bovis seu Bubali cream
0.5%th, yeast powder 0.5%, dipotassium hydrogen phosphate 0.02%, sodium hydroxide 0.05%, balance of water, pH7.0-7.3.
Step（1）In, condition of culture is：38.5 DEG C of culture 12-16 hours.
Step（2）In, it is 12-16 hour that condition of culture is 38.5 DEG C of temperature, pH7.0-7.3, incubation time.
Step（2）In, tank pressure 0.05MPa.
Step（3）In, fermentation tank tank pressure 0.05MP.
Step（3）Specifically, after being passed through aseptic oxygen-containing air 37-40 DEG C fermentation 18-32 hour, being passed through 39-45 DEG C of nitrogen
Continue fermentation, be passed through the nitrogen stage be simultaneously introduced concentration for 800g/L glucose 300mL, fermentation liquid pH fall per minute
Terminate fermentation during less than 0.01, fermentation broth viscosity is detected, fermentation liquid is pressed in settling tank.
Beneficial effects of the present invention are：
The present invention filters out the high strain of hyaluronic acid productivity, and in conjunction with improved fermentation technique, hyaluronic acid product output capacity is high,
Up to 9.5g/L, with short production cycle.Simple production process of the present invention, supplementary material is common to be easy to get, and produces easily controllable, impurity content
Lower, suitable industrialization large-scale production, high income, and greatly reduce production cost.
Specific embodiment：
For a better understanding of the present invention, technical scheme is described in detail with instantiation below, but the present invention
It is not limited thereto.
Embodiment 1
A kind of Malian drainage（Streptococcus equi subsp. zooepidemicus）DCS005, at present
The Malian drainage is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, address：Beijing
The institute 3 of city Chaoyang District North Star West Road 1, postcode：100101, its deposit number is：CGMCC No.12782, the Latin of strain
Title isStreptococcus equi subsp. zooepidemicus, join the microorganism of evidence（Strain）：DCS005, preservation day
Phase is on July 14th, 2016.
The production technology of hyaluronic acid is prepared with described Malian drainage, is comprised the following steps：
（1）Shake-flask seed liquid culture；
（2）Strain expanded culture；
（3）Fermentation, is first passed through the fermentation of aseptic oxygen-containing air, then is passed through nitrogen continuation fermentation；
（4）Purification.
Step（1）、（2）、（3）In culture fluid include starch 1% by weight percentage, sucrose 0.5%, Carnis Bovis seu Bubali cream
0.5%th, yeast powder 0.5%, dipotassium hydrogen phosphate 0.02%, sodium hydroxide 0.05%, balance of water, pH7.0-7.3.
Step（1）In, condition of culture is：38.5 DEG C of culture 12-16 hours.
Step（2）In, it is 12-16 hour that condition of culture is 38.5 DEG C of temperature, pH7.0-7.3, incubation time.
Step（2）In, tank pressure 0.05MPa.
Step（3）In, fermentation tank tank pressure 0.05MP.
Step（3）Specifically, after being passed through aseptic oxygen-containing air 37-40 DEG C fermentation 18-32 hour, being passed through 39-45 DEG C of nitrogen
Continue fermentation, be passed through the nitrogen stage be simultaneously introduced concentration for 800g/L glucose 300mL, fermentation liquid pH fall per minute
Terminate fermentation during less than 0.01, fermentation broth viscosity is detected, fermentation liquid is pressed in settling tank.
The present invention strain high using hyaluronic acid productivity is filtered out, combining with fermentation technology, hyaluronic acid product yield reaches
9.5g/L, with short production cycle.
Simple production process of the present invention, supplementary material is common to be easy to get, and production is easily controllable, and impurity content is lower, is suitable for industry
Change large-scale production, high income, and greatly reduce production cost.
Claims (8)
1. a kind of Malian drainage（Streptococcus equi subsp. zooepidemicus）DCS005, its
In China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, its deposit number is：CGMCC No.12782,
Preservation date is on July 14th, 2016.
2. the production technology of hyaluronic acid is prepared with the Malian drainage described in claim 1, it is characterised in that included
Following steps：
（1）Shake-flask seed liquid culture；
（2）Strain expanded culture；
（3）Fermentation, is first passed through the fermentation of aseptic oxygen-containing air, then is passed through nitrogen continuation fermentation；
（4）Purification.
3. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（1）、（2）、（3）In culture fluid include starch 1% by weight percentage, sucrose 0.5%, Carnis Bovis seu Bubali cream 0.5%, yeast powder
0.5%th, dipotassium hydrogen phosphate 0.02%, sodium hydroxide 0.05%, balance of water, pH7.0-7.3.
4. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（1）In, condition of culture is：38.5 DEG C of culture 12-16 hours.
5. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（2）In, it is 12-16 hour that condition of culture is 38.5 DEG C of temperature, pH7.0-7.3, incubation time.
6. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（2）In, tank pressure 0.05MPa.
7. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（3）In, fermentation tank tank pressure 0.05MP.
8. Malian drainage according to claim 2 prepares the production technology of hyaluronic acid, it is characterised in that step
Suddenly（3）Specifically, after being passed through aseptic oxygen-containing air 37-40 DEG C fermentation 18-32 hour, being passed through 39-45 DEG C of nitrogen and continuing fermentation, lead to
Enter the nitrogen stage be simultaneously introduced concentration for 800g/L glucose 300mL, fermentation liquid pH fall per minute less than 0.01 when
Terminate fermentation, fermentation broth viscosity is detected, fermentation liquid is pressed in settling tank.
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| CN201610844980.2A CN106434444B (en) | 2016-09-23 | 2016-09-23 | A kind of Malian drainage and its production technology for preparing hyaluronic acid |
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| CN201610844980.2A CN106434444B (en) | 2016-09-23 | 2016-09-23 | A kind of Malian drainage and its production technology for preparing hyaluronic acid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107338201A (en) * | 2017-06-23 | 2017-11-10 | 浙江工业大学 | Malian drainage SXY36 and the application in fermenting and producing hyaluronic acid |
| CN114806938A (en) * | 2022-04-20 | 2022-07-29 | 齐鲁工业大学 | Streptococcus equi subsp zooepidemicus for producing hyaluronic acid in low-sugar environment and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1102435A (en) * | 1994-08-17 | 1995-05-10 | 桂平县生物技术开发公司 | Method for producing sodium hyaluronate by microbial fermentation |
| CN101643760A (en) * | 2009-09-04 | 2010-02-10 | 四川理工学院 | Method for increasing the yield of hyaluronic acid |
| CN104059865A (en) * | 2014-06-11 | 2014-09-24 | 滨州安华生物工程有限公司 | Streptococcus zooepidemicus and production process for preparing hyaluronic acid by using same |
-
2016
- 2016-09-23 CN CN201610844980.2A patent/CN106434444B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1102435A (en) * | 1994-08-17 | 1995-05-10 | 桂平县生物技术开发公司 | Method for producing sodium hyaluronate by microbial fermentation |
| CN101643760A (en) * | 2009-09-04 | 2010-02-10 | 四川理工学院 | Method for increasing the yield of hyaluronic acid |
| CN104059865A (en) * | 2014-06-11 | 2014-09-24 | 滨州安华生物工程有限公司 | Streptococcus zooepidemicus and production process for preparing hyaluronic acid by using same |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107338201A (en) * | 2017-06-23 | 2017-11-10 | 浙江工业大学 | Malian drainage SXY36 and the application in fermenting and producing hyaluronic acid |
| CN107338201B (en) * | 2017-06-23 | 2021-02-02 | 浙江工业大学 | Streptococcus equi subsp zooepidemicus SXY36 and application thereof in fermentation production of hyaluronic acid |
| CN114806938A (en) * | 2022-04-20 | 2022-07-29 | 齐鲁工业大学 | Streptococcus equi subsp zooepidemicus for producing hyaluronic acid in low-sugar environment and application thereof |
| CN114806938B (en) * | 2022-04-20 | 2023-05-09 | 齐鲁工业大学 | Streptococcus equi subspecies zooepidemicus for producing hyaluronic acid in low-sugar environment and application thereof |
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| CN106434444B (en) | 2017-12-08 |
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Effective date of registration: 20200831 Address after: 257000 No.75 Huaihe Road, Dongying Economic Development Zone, Shandong Province Patentee after: DONGYING FOSTER BIOLOGICAL ENGINEERING Co.,Ltd. Address before: 257000, Shandong, Dongying, Kenli Town, West Tuo (Yong Xin Road, No. 98) Co-patentee before: DONGYING FOSTER BIOLOGICAL ENGINEERING Co.,Ltd. Patentee before: CHINA EASTAR HOLDINGS GROUP Co.,Ltd. |
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