CN113801910A - Preparation method of pure natural riboflavin - Google Patents
Preparation method of pure natural riboflavin Download PDFInfo
- Publication number
- CN113801910A CN113801910A CN202111001697.0A CN202111001697A CN113801910A CN 113801910 A CN113801910 A CN 113801910A CN 202111001697 A CN202111001697 A CN 202111001697A CN 113801910 A CN113801910 A CN 113801910A
- Authority
- CN
- China
- Prior art keywords
- riboflavin
- percent
- culture
- fermentation
- pure natural
- 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
Classifications
-
- 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
- C12P25/00—Preparation of compounds containing alloxazine or isoalloxazine nucleus, e.g. riboflavin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D475/00—Heterocyclic compounds containing pteridine ring systems
- C07D475/12—Heterocyclic compounds containing pteridine ring systems containing pteridine ring systems condensed with carbocyclic rings or ring systems
- C07D475/14—Benz [g] pteridines, e.g. riboflavin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of riboflavin production by a fermentation method, in particular to a preparation method of pure natural riboflavin, which adopts Eremothecium ashbyii and does not undergo any gene recombination, mutagenesis, construction or reconstruction, the obtained finished riboflavin product completely meets the requirements of national non-transgenic product laws and regulations, the obtained riboflavin product can meet the standard regulations of EP10.0 and Chinese pharmacopoeia CP2020, and the riboflavin product is in a spherical crystal form and has good processing performance.
Description
Technical Field
The invention belongs to the technical field of riboflavin production by a fermentation method, and particularly relates to a preparation method of pure natural riboflavin.
Background
Riboflavin also known as vitamin B2Is a kind of coreThe sugar alcohol group isoalloxazine derivatives, which are prepared by various microbial and plant biosyntheses.
At present, the method for mainly producing riboflavin is a microbial fermentation method, and the microorganism suitable for the production by the fermentation method may be any riboflavin-producing microorganism existing in the natural world or riboflavin-overproducing microorganism transformed by genetic engineering, chemical or physical methods. These microorganisms are cultured under appropriate conditions to produce riboflavin. The produced riboflavin is recovered from the culture.
European patent No. EP0821063 discloses a method for producing riboflavin using recombinant Bacillus subtilis. U.S. Pat. No.5837528 discloses a recombinant strain overproducing riboflavin of Bacillus subtilis, which is obtained by introducing the rib operon into the parent strain by recombinant techniques. There are also reports by WindhoIz of Eremothecium ashbyii and Ashbya gossypii ascomycete as riboflavin-producing microorganisms, but these methods all have the common disadvantage: the production strain must be subjected to gene recombination, mutagenesis, construction or modification, so that the production process of the riboflavin is not purely natural, and the existing market gradually rejects the riboflavin produced by the gene recombination of the strain, so that the demand for preparing the riboflavin produced by the purely natural strain without gene modification is increasing.
However, the production of riboflavin by using pure natural strains without gene modification has been an industrial problem how to make the quality standard meet the impurity limit requirements of pharmacopoeia standards of various countries, especially under the background of further prompting the requirements of the Chinese pharmacopoeia CP2020 on riboflavin standards.
Disclosure of Invention
In order to solve the problems, the invention provides a preparation method for producing riboflavin by using purely natural strains without gene modification, which meets the national non-transgenic product law and regulation requirements, simultaneously the obtained riboflavin product can meet the standard regulations of EP10.0 and Chinese pharmacopoeia CP2020, and the riboflavin product is in a spherical crystal form and has good processing performance.
The specific technical scheme is a preparation method of pure natural riboflavin, which is characterized by comprising the following steps:
step 1, inoculating the eremothecium ashbyii in a shake flask culture medium for metabolic culture to obtain a spore suspension containing riboflavin;
step 2, enabling the spore suspension to enter a seeding tank through the micropores, and carrying out metabolic culture in a seed culture medium to obtain a seed solution;
step 3, transferring the seed liquid into a fermentation tank, and carrying out metabolic culture in a fermentation culture medium to obtain fermentation liquid containing riboflavin;
step 4, dissolving the fermentation liquor by using alkali, adjusting the pH value to 11.0-12.0, stirring and dissolving the riboflavin, and filtering and washing to obtain filtrate containing the riboflavin;
step 5, adding inorganic acid into the filtrate containing riboflavin to adjust the pH value to 5.0-6.0, standing to crystallize and separate out the riboflavin from the solution, and obtaining a crystallization liquid containing riboflavin crystals;
step 6, filtering and concentrating the crystal liquid containing the riboflavin crystals to obtain concentrated liquid containing the riboflavin;
and 7, spray drying the concentrated solution containing the riboflavin to obtain a spherical crystal form riboflavin finished product.
And in the step 1, the components of the shake-flask culture medium at least comprise 3.5-4.0% of vegetable oil, 0.5% of maltose, 2% of bone glue, 2-3% of corn steep liquor, 1% of fish meal, 0.2% of sodium chloride, 0.1% of calcium chloride, 0.1% of potassium dihydrogen phosphate and 0.1% of magnesium sulfate, and the culture temperature of the shake-flask culture medium is 28-29 ℃ and the culture time is 3-5 days.
In step 2, the components of the seed culture medium at least comprise 2 percent of vegetable oil, 1 to 2 percent of glucose, 2 percent of bone glue, 2 to 3 percent of corn steep liquor, 0.5 percent of peptone, 0.2 percent of sodium chloride, 0.1 percent of potassium dihydrogen phosphate, 0.5 percent of maltose, 0.1 percent of calcium chloride and 0.1 percent of magnesium sulfate, and the culture temperature of the seed culture medium is 29 to 31 ℃ and the culture time is 40 hours.
And in the step 3, the components of the fermentation medium at least comprise 2 percent of vegetable oil, 1 to 2 percent of glucose, 2 percent of bone glue, 2 to 3 percent of corn steep liquor, 0.5 percent of peptone, 0.2 percent of sodium chloride, 0.1 percent of monopotassium phosphate and less than 0.1 percent of defoaming agent, and the culture condition of the fermentation medium is that the culture temperature is 28 to 30 ℃ and the culture time is 160 hours.
Furthermore, the base in step 4 is a 30% sodium hydroxide solution.
In step 4, ceramic membrane, multistage microfiltration and ultrafiltration membrane separation equipment are adopted for filtration and washing, the pore diameter of the membrane is 10-500nm, the temperature in the whole process is controlled below 25 ℃, and washing liquid used for washing is sodium hydroxide solution with the pH value of 11.0-12.0 and water.
And in the step 5, the inorganic acid is hydrochloric acid and/or hydrogen peroxide.
In step 6, ceramic membrane, multistage microfiltration and ultrafiltration membrane separation equipment are adopted for filtration and concentration, the pore diameter of the membrane is 100nm-1 μm, and the temperature in the whole process is controlled below 35 ℃.
The beneficial effects of this technical scheme lie in:
the eremothecium ashbyii is derived from Shanghai plant institute of Chinese academy of sciences and is not subjected to any gene recombination, mutagenesis, construction or transformation, the obtained riboflavin finished product completely meets the national non-transgenic product law and regulation requirements, the product quality reaches the requirements of pharmacopoeia of various countries, and the impurities are low.
Secondly, the problems of high viscosity and difficult extraction of fermentation broth obtained by fermenting Eremothecium ashbyii (Eremothecium ashbyii) bacteria in the prior art are solved by optimizing the formula of the culture medium.
Thirdly, the problems of long process and serious environmental pollution of the traditional extraction process (2-hydroxy-3-naphthoic acid method) of the fermentation liquor of the Eremothecium ashbyii are solved.
The riboflavin obtained by the method is spherical crystalline riboflavin, has good processing performance and good fluidity, is convenient to apply and improves the quality of the riboflavin product.
Fifthly, the high-purity finished riboflavin product is directly extracted and prepared from the fermentation liquor, thereby reducing sewage discharge, shortening the extraction process and avoiding the degradation and loss of the riboflavin in the extraction process.
Detailed Description
The present invention will be described in detail with reference to examples, but the present invention is not limited to the examples.
Example 1
(1) Eremothecium ashbyii, from Haematococcus Shanghai, academy of sciences, was used as parent strain (parent strain refers to strain that has not been mutagenized) to prepare spore suspension: inoculating the Erigerus albugineus in a shake flask, culturing in a shake flask culture medium at 28-29 deg.C for 3-5 days. The shake flask culture medium uses a mixed carbon source and a mixed nitrogen source as main culture media, and a small amount of inorganic salt is added. The shake culture medium per liter at least contains 3.5% vegetable oil, 0.5% maltose, 2% bone glue, 2.5% corn steep liquor, 1% fish meal, 0.2% sodium chloride, 0.1% calcium chloride, 0.1% monopotassium phosphate, and 0.1% magnesium sulfate. Larger colonies were picked for spore suspension.
(2) Seed liquor was cultured in a seed tank by fermentation with a parent strain of Eremothecium ashbyii:
and (2) enabling the spore suspension obtained in the step (1) to flow into a seeding tank through pressure change by adopting a micropore inoculation method, wherein the culture temperature of a seed culture medium is 29-31 ℃, the culture time is 40 hours, and the normal growth and propagation of the seeds are maintained under the conditions of ventilation and stirring. The seed culture medium takes a mixed carbon source and a mixed nitrogen source as main culture media, and is added with a small amount of inorganic salt, and each liter of the culture medium at least contains 2 percent of vegetable oil, 1 percent of glucose, 2 percent of bone glue, 2 percent of corn steep liquor, 0.5 percent of peptone, 0.2 percent of sodium chloride, 0.1 percent of potassium dihydrogen phosphate, 0.5 percent of maltose, 0.1 percent of calcium chloride and 0.1 percent of magnesium sulfate. And (5) preparing for seed transfer when the concentration of the strain reaches 10-15% of the whole volume of the seed culture medium.
(3) Preparing fermentation liquor: a method for producing riboflavin by fermenting and culturing parent strains of Eremothecium ashbyii in a fermenter comprises the following steps:
the level of riboflavin production by the Eremothecium ashbyii strain was determined by fermentation culture of a parent strain of Eremothecium ashbyii in a 6T fermentor. Transferring the seed liquid obtained in the step (2) into a fermentation tank according to the seed transferring amount of 10-15%, and fermenting and culturing at 28-30 ℃ for 160 hours. Preventing pollution in the fermentation culture process. The fermentation medium takes a mixed carbon source and a mixed nitrogen source as main culture media, and a small amount of inorganic salt is added, wherein each liter of the culture medium at least contains 2% of vegetable oil, 1% of glucose, 2% of bone glue, 3% of corn steep liquor, 0.5% of peptone, 0.2% of sodium chloride, 0.1% of monopotassium phosphate and a small amount of antifoaming agent. The pH was adjusted to 6.4 with sodium hydroxide. And (5) preparing to put the tank after the fermentation is finished.
(4) At the end of the fermentation, the productivity of riboflavin from the parent strain of Eremothecium ashbyii was measured by HPLC. A chromatographic column using octadecylsilane chemically bonded silica as a filler and 0.01mol/L sodium heptanesulfonate 0.5 percent glacial acetic acid solution-acetonitrile-methanol (ratio 85: 10: 5) as a mobile phase; measured at a wavelength of 444 nm. According to the experimental result, the riboflavin production capacity of the strain reaches the actual production requirement, and the strain has actual production value.
(5) Extraction of riboflavin from the fermentation broth: 30% sodium hydroxide solution was added to the 4T riboflavin broth, the pH was adjusted to 11.5, and the mixture was stirred at 25 ℃ to dissolve the riboflavin. Then feeding the feed liquid into a multistage continuous membrane filtration device for impurity removal, wherein ceramic membrane, multistage microfiltration and ultrafiltration membrane separation devices can be adopted. The pore diameter of the membrane is 100nm and 500nm, and the operation temperature is controlled below 25 ℃. Adding hydrochloric acid into the multistage continuous membrane filtrate to adjust the pH value to 5.5-6.0, and then adding hydrogen peroxide with the volume of 0.2% of the feed liquid. Stirring for 30min, standing for 12 hr to precipitate riboflavin crystals, and concentrating the feed liquid containing riboflavin crystals in multi-stage continuous membrane equipment to separate crystals. The pore diameter of all the membranes is 200nm-800nm, and the temperature of feed liquid is maintained below 35 ℃ in the process. Spray drying the riboflavin concentrate to obtain spherical crystalline riboflavin granules. Through analysis, the product quality reaches the requirements of riboflavin EP10.0 and riboflavin CP2020 version pharmacopoeia.
TABLE 1 analysis of impurities in finished riboflavin product obtained in example 1
Example 2
(1) Inoculating the Erigerus albugineus in a shake flask, culturing in a shake flask culture medium at 28-29 deg.C for 3-5 days. The shake flask culture medium uses a mixed carbon source and a mixed nitrogen source as main culture media, and a small amount of inorganic salt is added. The shake culture medium per liter at least contains 3.5% vegetable oil, 0.5% maltose, 2% bone glue, 2% corn steep liquor, 1% fish meal, 0.2% sodium chloride, 0.1% calcium chloride, 0.1% potassium dihydrogen phosphate, and 0.1% magnesium sulfate. Larger colonies were picked for spore suspension.
(2) Seed liquor was cultured in a seed tank by fermentation with a parent strain of Eremothecium ashbyii:
and (2) enabling the spore suspension obtained in the step (1) to flow into a seeding tank by adopting a micropore inoculation method through pressure change, culturing the seeds at the temperature of 29-31 ℃ for 40 hours, and maintaining the normal growth and propagation of the seeds under the conditions of ventilation and stirring. The seed culture medium takes a mixed carbon source and a mixed nitrogen source as main culture media, and is added with a small amount of inorganic salt, and each liter of the culture medium at least contains 2 percent of vegetable oil, 2 percent of glucose, 2 percent of bone glue, 2 percent of corn steep liquor, 0.5 percent of peptone, 0.2 percent of sodium chloride, 0.1 percent of potassium dihydrogen phosphate, 0.5 percent of maltose, 0.1 percent of calcium chloride and 0.1 percent of magnesium sulfate. And (5) preparing for transplanting when the bacteria concentration reaches a certain value.
(3) Preparing fermentation liquor: a method for producing riboflavin by fermenting and culturing parent strains of Eremothecium ashbyii in a fermenter comprises the following steps:
the level of riboflavin production capacity of the Eremothecium ashbyii strain was determined by fermentation culture of the parent strain of Eremothecium ashbyii in a 30T fermentor (scale-up production further embodies the stability of the protocol). Transferring the seed liquid obtained in the step (2) into a fermentation tank according to the seed transferring amount of 10-15%, and fermenting and culturing at 28-30 ℃ for 160 hours. Preventing pollution in the fermentation culture process. The fermentation medium takes a mixed carbon source and a mixed nitrogen source as main culture media, and a small amount of inorganic salt is added, wherein each liter of the culture medium at least contains 2% of vegetable oil, 2% of glucose, 2% of bone glue, 2% of corn steep liquor, 0.5% of peptone, 0.2% of sodium chloride, 0.1% of monopotassium phosphate and a small amount of antifoaming agent. The pH was adjusted to 6.4 with sodium hydroxide. And (5) preparing to put the tank after the fermentation is finished.
(4) At the end of the fermentation, the productivity of riboflavin from the parent strain of Eremothecium ashbyii was measured by HPLC. A chromatographic column using octadecylsilane chemically bonded silica as a filler and 0.01mol/L sodium heptanesulfonate 0.5 percent glacial acetic acid solution-acetonitrile-methanol (ratio 85: 10: 5) as a mobile phase; measured at a wavelength of 444 nm. According to the experimental result, the capacity of producing riboflavin of the strain reaches the actual production requirement. Has practical production value.
(5) Extraction of riboflavin from the fermentation broth: 30 percent of alkali liquor is added into 30T of riboflavin fermentation liquor, the pH is adjusted to be 11.8, and the riboflavin is dissolved by stirring under the condition of 25 ℃. Then feeding the feed liquid into a multistage continuous membrane filtration device for impurity removal, wherein ceramic membrane, multistage microfiltration and ultrafiltration membrane separation devices can be adopted. The membrane aperture is 50-300nm, and the operation temperature is controlled below 25 ℃. Adding hydrochloric acid into the multistage continuous membrane filtrate to adjust the pH value to 5.8-6.0, and then adding hydrogen peroxide with the volume of 0.3% of the feed liquid. Stirring for 30min, standing for 12 hr to precipitate riboflavin crystals, and concentrating the feed liquid containing riboflavin crystals in multi-stage continuous membrane equipment to separate crystals. The pore size of all membranes is 100nm-900nm, and the temperature of feed liquid is maintained below 35 ℃ in the process. Spray drying the riboflavin concentrate. Spherical crystalline riboflavin particles are obtained. The product quality reaches the requirements of riboflavin EP10.0 and riboflavin CP2020 version pharmacopoeia.
Claims (8)
1. A preparation method of pure natural riboflavin is characterized by comprising the following preparation steps:
step 1, inoculating the eremothecium ashbyii in a shake flask culture medium for metabolic culture to obtain a spore suspension containing riboflavin;
step 2, enabling the spore suspension to enter a seeding tank through the micropores, and carrying out metabolic culture in a seed culture medium to obtain a seed solution;
step 3, transferring the seed liquid into a fermentation tank, and carrying out metabolic culture in a fermentation culture medium to obtain fermentation liquid containing riboflavin;
step 4, dissolving the fermentation liquor by using alkali, adjusting the pH value to 11.0-12.0, stirring and dissolving the riboflavin, and filtering and washing to obtain filtrate containing the riboflavin;
step 5, adding inorganic acid into the filtrate containing riboflavin to adjust the pH value to 5.0-6.0, standing to crystallize and separate out the riboflavin from the solution, and obtaining a crystallization liquid containing riboflavin crystals;
step 6, filtering and concentrating the crystal liquid containing the riboflavin crystals to obtain concentrated liquid containing the riboflavin;
and 7, spray drying the concentrated solution containing the riboflavin to obtain a spherical crystal form riboflavin finished product.
2. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 1, the components of the shake-flask culture medium at least comprise 3.5-4.0% of vegetable oil, 0.5% of maltose, 2% of bone glue, 2-3% of corn steep liquor, 1% of fish meal, 0.2% of sodium chloride, 0.1% of calcium chloride, 0.1% of potassium dihydrogen phosphate and 0.1% of magnesium sulfate, and the culture temperature of the shake-flask culture medium is 28-29 ℃ and the culture time is 3-5 days.
3. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 2, the components of the seed culture medium at least comprise 2 percent of vegetable oil, 1 to 2 percent of glucose, 2 percent of bone glue, 2 to 3 percent of corn steep liquor, 0.5 percent of peptone, 0.2 percent of sodium chloride, 0.1 percent of potassium dihydrogen phosphate, 0.5 percent of maltose, 0.1 percent of calcium chloride and 0.1 percent of magnesium sulfate, and the culture condition of the seed culture medium is that the culture temperature is 29 to 31 ℃ and the culture time is 40 hours.
4. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 3, the fermentation medium at least contains 2% of vegetable oil, 1% -2% of glucose, 2% of bone glue, 2% -3% of corn steep liquor, 0.5% of peptone, 0.2% of sodium chloride, 0.1% of monopotassium phosphate and less than 0.1% of defoaming agent, and the culture conditions of the fermentation medium are that the culture temperature is 28-30 ℃ and the culture time is 160 hours.
5. The process for preparing pure natural riboflavin according to claim 1, wherein: the base in step 4 is 30% sodium hydroxide solution.
6. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 4, ceramic membrane, multistage microfiltration and ultrafiltration membrane separation equipment are adopted for filtration and washing, the pore diameter of the membrane is 10-500nm, the temperature in the whole process is controlled below 25 ℃, and washing liquid used for washing is sodium hydroxide solution with the pH value of 11.0-12.0 and water.
7. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 5, the inorganic acid is hydrochloric acid and/or hydrogen peroxide.
8. The process for preparing pure natural riboflavin according to claim 1, wherein: in the step 6, ceramic membrane, multistage microfiltration and ultrafiltration membrane separation equipment are adopted for filtration and concentration, the aperture of the membrane is 100nm-1 mu m, and the temperature in the whole process is controlled below 35 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111001697.0A CN113801910A (en) | 2021-08-30 | 2021-08-30 | Preparation method of pure natural riboflavin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111001697.0A CN113801910A (en) | 2021-08-30 | 2021-08-30 | Preparation method of pure natural riboflavin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113801910A true CN113801910A (en) | 2021-12-17 |
Family
ID=78894335
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111001697.0A Pending CN113801910A (en) | 2021-08-30 | 2021-08-30 | Preparation method of pure natural riboflavin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113801910A (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB623082A (en) * | 1942-03-16 | 1949-05-11 | Commercial Solvents Corp | Improvements in or relating to a process for the production of riboflavin |
| EP0338596A1 (en) * | 1983-09-09 | 1989-10-25 | Daicel Chemical Industries, Ltd. | Process for the preparation of riboflavin |
| CN1049185A (en) * | 1989-06-22 | 1991-02-13 | 霍夫曼-拉罗奇有限公司 | The bacterial isolates of Riboflavinoverproducstrains |
| US5210023A (en) * | 1990-07-04 | 1993-05-11 | Basf Aktiengesellschaft | Method of purifying ferment-produced riboflavin |
| CN1232872A (en) * | 1998-04-23 | 1999-10-27 | 弗·哈夫曼-拉罗切有限公司 | Overproduction of riboflavin in yeast |
| CN1639353A (en) * | 2002-03-02 | 2005-07-13 | 巴斯福股份公司 | Method for producing riboflavin |
| CN101205230A (en) * | 2007-12-10 | 2008-06-25 | 南京九思高科技有限公司 | Method for extracting high-purity riboflavin directly from fermentation liquor |
| CN104961740A (en) * | 2015-06-02 | 2015-10-07 | 湖北广济药业股份有限公司 | Riboflavin-containing solid preparation and preparation method thereof |
| CN105483071A (en) * | 2015-12-29 | 2016-04-13 | 天津大学 | High-yield riboflavin Escherichia coli engineering strain, and construction and fermentation method thereof |
| CN106957129A (en) * | 2017-03-30 | 2017-07-18 | 湖北广济药业股份有限公司 | A kind of processing method of Lactochrome fermentation liquor |
| CN108913747A (en) * | 2018-07-28 | 2018-11-30 | 广济药业（孟州）有限公司 | A kind of high density fermentation vitamin B2Method |
| CN111334552A (en) * | 2020-03-03 | 2020-06-26 | 河南巨龙生物工程股份有限公司 | Fermentation production method of riboflavin |
-
2021
- 2021-08-30 CN CN202111001697.0A patent/CN113801910A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB623082A (en) * | 1942-03-16 | 1949-05-11 | Commercial Solvents Corp | Improvements in or relating to a process for the production of riboflavin |
| EP0338596A1 (en) * | 1983-09-09 | 1989-10-25 | Daicel Chemical Industries, Ltd. | Process for the preparation of riboflavin |
| CN1049185A (en) * | 1989-06-22 | 1991-02-13 | 霍夫曼-拉罗奇有限公司 | The bacterial isolates of Riboflavinoverproducstrains |
| US5210023A (en) * | 1990-07-04 | 1993-05-11 | Basf Aktiengesellschaft | Method of purifying ferment-produced riboflavin |
| CN1232872A (en) * | 1998-04-23 | 1999-10-27 | 弗·哈夫曼-拉罗切有限公司 | Overproduction of riboflavin in yeast |
| CN1639353A (en) * | 2002-03-02 | 2005-07-13 | 巴斯福股份公司 | Method for producing riboflavin |
| CN101205230A (en) * | 2007-12-10 | 2008-06-25 | 南京九思高科技有限公司 | Method for extracting high-purity riboflavin directly from fermentation liquor |
| CN104961740A (en) * | 2015-06-02 | 2015-10-07 | 湖北广济药业股份有限公司 | Riboflavin-containing solid preparation and preparation method thereof |
| CN105483071A (en) * | 2015-12-29 | 2016-04-13 | 天津大学 | High-yield riboflavin Escherichia coli engineering strain, and construction and fermentation method thereof |
| CN106957129A (en) * | 2017-03-30 | 2017-07-18 | 湖北广济药业股份有限公司 | A kind of processing method of Lactochrome fermentation liquor |
| CN108913747A (en) * | 2018-07-28 | 2018-11-30 | 广济药业（孟州）有限公司 | A kind of high density fermentation vitamin B2Method |
| CN111334552A (en) * | 2020-03-03 | 2020-06-26 | 河南巨龙生物工程股份有限公司 | Fermentation production method of riboflavin |
Non-Patent Citations (3)
| Title |
|---|
| A.E KALINGAN: "Influence of type and concentration of flavinogenic factors on production of riboflavin by Eremothecium ashbyii NRRL 1363", 《BIORESOURCE TECHNOLOGY》, vol. 82, no. 3, pages 219 - 224 * |
| TIJEN ÖZBAS等: "Comparative study of riboflavin production from two microorganisms: Eremothecium ashbyii and Ashbya gossypii", 《ENZYME AND MICROBIAL TECHNOLOGY》, vol. 8, no. 10, pages 593 - 596 * |
| 李嵘等: "发酵法制核黄素", 《无锡轻工大学学报》, vol. 16, no. 1, pages 14 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11319561B2 (en) | Method for producing a long chain dicarboxylic acid by fermentation, fermentation broth, treated fermentation broth and wastewater | |
| CN111748480B (en) | Candida virginiana and application thereof | |
| CN104893983B (en) | Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product | |
| EP4299578A1 (en) | Method for co-producing erythritol and arabinose from xylose mother liquor | |
| CN102061320B (en) | Preparation method of 11 alpha,17 alpha-dyhydroxyl-androst-4-ene-3,20-dione | |
| CN114634952A (en) | Method for improving erythritol yield and sugar alcohol conversion rate | |
| CN113321580A (en) | Method for producing malic acid | |
| CN107058414B (en) | Method for preparing L-alanine | |
| CN106834377B (en) | Method for producing epothilone B | |
| CN106434823A (en) | Fermentation method of CPC (cephalosporin C) | |
| CN102703334B (en) | Strain producing erythritol and method for producing erythritol by using strain | |
| CN114249645B (en) | A method for extracting sebacic acid and sebacic acid product | |
| CN113801910A (en) | Preparation method of pure natural riboflavin | |
| CN117126898B (en) | Process for preparing valine by biotechnology | |
| CN116179356B (en) | Method for high-density heterotrophic culture of chlamydomonas reinhardtii and application thereof | |
| CN106854670B (en) | Method for producing teicoplanin and regulating and controlling component content of teicoplanin by fermentation method | |
| CN111100823B (en) | Polymyxin B sulfate production strain, preparation method and application of polymyxin B sulfate | |
| CN107177508B (en) | Method for synthesizing and producing long-chain dodecanedioic acid by biological method | |
| CN110592154B (en) | Process for producing and extracting tryptophan | |
| CN116218747A (en) | Streptomyces avermitilis fermentation medium based on biomass and fermentation method | |
| CN115678925A (en) | Preparation method of calcium propionate | |
| CN107674854B (en) | Nitrogen-fixing sphingosine monad and application thereof in preparation of gellan gum | |
| CN102618468B (en) | Temperature-resistant alcaligenes and application thereof in welan gum production | |
| CN112592945A (en) | Adenosine fermentation process | |
| CN111635917A (en) | Preparation method of beta-nicotinamide ribodinucleotide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211217 |
|
| RJ01 | Rejection of invention patent application after publication |