CN113308500A - Method for extracting vitamin K2 from fermentation liquor - Google Patents

Method for extracting vitamin K2 from fermentation liquor Download PDF

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CN113308500A
CN113308500A CN202110689781.XA CN202110689781A CN113308500A CN 113308500 A CN113308500 A CN 113308500A CN 202110689781 A CN202110689781 A CN 202110689781A CN 113308500 A CN113308500 A CN 113308500A
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vitamin
acetone
thalli
extracting
fermentation
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卢伟
张倩
马善丽
徐艳
吴祥舟
李飞
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Shandong Runde Biotechnology Co Ltd
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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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/66—Preparation of oxygen-containing organic compounds containing the quinoid structure
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C46/00—Preparation of quinones
    • C07C46/10—Separation; Purification; Stabilisation; Use of additives
    • 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
    • C12N1/20—Bacteria; Culture media therefor

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Abstract

The invention relates to the technical field of vitamin K2 preparation, in particular to a method for extracting vitamin K2 from fermentation liquor, which comprises the following steps: (1) inoculating flavobacterium to a culture medium for fermentation, adding a sedimentation promoting agent into fermentation liquor after the fermentation is finished, and separating out thalli for later use after the thalli in the fermentation liquor are settled. (2) Freeze-drying the thalli, crushing the thalli into powder, mixing the powder with acetone, extracting, and taking an extract for later use. (3) And adding clear water into the extract, and separating out precipitates after precipitation of the precipitates is finished to obtain a crude product of the vitamin K2 product. (4) And mixing the vitamin K2 product crude product with acetone again for extraction, then adding clear water, and separating out precipitate after the precipitate is separated out to obtain the vitamin K2 product. The method of the invention not only can effectively extract vitamin K2 in flavobacterium, but also has the characteristics of convenient extraction and suitability for industrial application.

Description

Method for extracting vitamin K2 from fermentation liquor
Technical Field
The invention relates to the technical field of vitamin K2 preparation, in particular to a method for extracting vitamin K2 from fermentation liquor.
Background
In 1943, Danish biochemist Dahm and American scientist Doixian discovered vitamins K1 and K2, and discovered that vitamin K2 has an important effect on bones, in 1995, Japan began to use the vitamin K2 preparation as a medicine for improving osteoporosis with osteopenia and pain, and with the continuous research on vitamin K2, scientists discovered that vitamin K2 not only can effectively prevent osteoporosis, but also can supplement proteins in the body and prevent cancer, and besides, has various effects of promoting the formation of prothrombin, accelerating blood coagulation, ensuring the normal blood coagulation, and effectively helping blood and bones to keep normal.
At present, the preparation method of vitamin K2 mainly comprises a chemical synthesis method and a microbial fermentation method, wherein, the extraction of metabolites in thallus by using flavobacterium after fermentation is a common preparation method of vitamin K2, for example, a method of extracting vitamin K2 in the thallus after breaking the wall of the thallus by using repeated freeze-thawing flavobacterium is adopted in some prior art, but the whole process of the method is complicated and is not suitable for industrial production and application.
Disclosure of Invention
Aiming at the problems, the invention provides a method for extracting vitamin K2 from fermentation liquor, which not only can effectively extract vitamin K2 from flavobacterium, but also has the characteristics of convenient extraction and suitability for industrial application. In order to achieve the purpose, the invention discloses the following technical scheme:
a method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating flavobacterium to a culture medium for fermentation, adding a sedimentation promoting agent into fermentation liquor after the fermentation is finished, and separating out thalli for later use after the thalli in the fermentation liquor are settled.
(2) Freeze-drying the thalli, crushing the thalli into powder, mixing the powder with acetone, extracting, and taking an extract for later use.
(3) And adding clear water into the extract, and separating out precipitates after precipitation of the precipitates is finished to obtain a crude product of the vitamin K2 product.
(4) And mixing the vitamin K2 product crude product with acetone again for extraction, then adding clear water, and separating out precipitate after the precipitate is separated out to obtain the vitamin K2 product.
Further, a culture medium with the following components is adopted in the step (1): 6-8.5 g/L of glycerol, 20-28 g/L of peptone and 2-3.5 g/L, K of yeast powder2HPO4 3～7.6g/L、NaCl 3～5g/L、MgSO40.1-4 g/L and 15-25 g/L of agar.
Further, in the step (1), the inoculation amount of the flavobacterium is 4-6.5%, the culture temperature is 24-28 ℃, the stirring speed is 250-350 r/min, and the culture time is 5-6 days.
Further, in the step (1), the precipitation accelerator includes any one of aluminum sulfate, ferric sulfate, aluminum chloride, ferric chloride, alum, polyaluminum chloride, polyferric chloride and the like. Optionally, the concentration of the sedimentation promoting agent in the fermentation liquid is 40-60 mg/L.
In step (1), the cells are separated by any of centrifugation, filtration and the like.
Further, in the step (2), the thalli is rapidly cooled to below-20 ℃, then the temperature is vacuumized to 0.1-0.2 mbra and the heating is started, and the freeze-dried thalli is obtained after water is sublimated.
Further, in the step (2), the volume mass ratio of the acetone to the powder is 10-15 ml: 1g, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to remove after the extraction is finished, so that the extraction liquid is obtained.
Further, in the step (4), the mass-to-volume ratio of the crude vitamin K2 product to acetone is 1 g: 8-13 ml, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to obtain the extraction liquid.
Further, in the steps (3) and (4), the addition amount of the clear water is more than 2 times, preferably 2-3.5 times of the volume of the acetone, and the clear water comprises any one of distilled water, purified water and the like. The vitamin K2 in the extract is forced out by adding clear water as the solvent of acetone.
Further, the method also comprises a step of distilling the residual extract liquid after the precipitate is separated out to recover acetone in the extract liquid, wherein the distillation temperature is higher than the boiling point of acetone and lower than the boiling point of water, and is preferably 60-65 ℃.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention adopts a method of directly processing the fermented thalli into freeze-dried thalli, and then adopts the propanol and the water for extraction, and the method not only conveniently and efficiently prepares the thalli containing the vitamin K2 into semi-finished products which are easy to store and sell, but also provides researchers to carry out tests of different extraction methods on the basis. Compared with the method which needs to realize the wall breaking of the thalli by a repeated freeze thawing method, the method can not cause the problem of product loss caused by the outflow of partial products in the ruptured thalli in the thawing process, and avoids unnecessary consumption of the products.
(2) The invention adopts a method of crushing the freeze-dried thallus, extracting by acetone, and then using clear water to force vitamin K2 in the extract to precipitate, because vitamin K2 can be largely dissolved in an organic solvent such as acetone, but is difficult to dissolve in water. Therefore, after the acetone is used as a solvent to extract the vitamin K2 in the crushed thallus, and then water with the volume larger than that of the acetone is added, the acetone is used as a solute to be quickly dissolved in the water, the vitamin K2 is difficult to be dissolved in the water together with the acetone, and then the vitamin K2 is precipitated from the water, and a vitamin K2 product with high purity is obtained after re-extraction. The method effectively simplifies the prior process for extracting the vitamin K2 from the fermentation liquor, reduces the cost and is more suitable for industrial application.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. The invention will now be further illustrated by specific examples.
Example 1
A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating the flavobacterium strain to a culture medium in a fermentation tank after seed culture, wherein the culture medium comprises the following components: 7g/L of glycerol, 25g/L of peptone and 3g/L, K of yeast powder2HPO4 5g/L、NaCl 3g/L、MgSO41g/L and 20g/L of agar, wherein the inoculation amount of the flavobacterium strain is 6 percent, the culture temperature is 25 ℃, the stirring speed is 250r/min, and the culture time is 6 days.
(2) After the culture is finished, alum is added into the fermentation liquor to enable the thalli to settle, the concentration of the sedimentation promoting agent in the fermentation liquor is 40mg/L, and after the thalli are completely settled, the thalli are filtered and separated for later use.
(3) And (3) placing the thalli in a freeze dryer, rapidly cooling to minus 25 ℃, then vacuumizing to 0.1mbra and starting heating, obtaining freeze-dried thalli after water is sublimated, and grinding the freeze-dried thalli into powder.
(4) Adding acetone into the powder, mixing and extracting, wherein the volume mass ratio of the acetone to the powder is 12 ml: 1g, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to remove after the extraction is finished, so that the extraction liquid is obtained.
(3) And (3) adding distilled water into the extract liquor obtained in the step (2), wherein the adding amount of the distilled water is 3 times of the volume of the acetone obtained in the step (2), stirring, standing, filtering and separating out a precipitate after the precipitate is separated out, and thus obtaining a crude product of the vitamin K2 product.
(4) Mixing the crude product of the vitamin K2 with acetone again, wherein the volume mass ratio of the acetone to the crude product is 13 ml: 1g, extracting for 35min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(5) And (4) adding distilled water into the extract liquor obtained in the step (4), wherein the adding amount of the distilled water is 2 times of the volume of the acetone obtained in the step (4), stirring, standing, and filtering to separate out a precipitate after the precipitate is separated out, so as to obtain the vitamin K2 product.
(6) The remaining extracts after the precipitates were separated in steps (3) and (5) were combined and then distilled at 65 ℃ to recover acetone.
Example 2
A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating the flavobacterium strain to a culture medium in a fermentation tank after seed culture, wherein the culture medium comprises the following components: 7g/L of glycerol, 25g/L of peptone and 3g/L, K of yeast powder2HPO4 5g/L、NaCl 3g/L、MgSO41g/L and 20g/L of agar, wherein the inoculation amount of the flavobacterium strain is 6.5 percent, the culture temperature is 24 ℃, the stirring speed is 350r/min, and the culture time is 5 days.
(2) And after the culture is finished, adding aluminum chloride into the fermentation liquor to settle the thalli, wherein the concentration of the sedimentation accelerator in the fermentation liquor is 60mg/L, and filtering and separating the thalli for later use after the thalli are completely settled.
(3) And (3) placing the thalli in a freeze dryer, rapidly cooling to-20 ℃, then vacuumizing to 0.15mbra and starting heating, obtaining freeze-dried thalli after water is sublimated, and grinding the freeze-dried thalli into powder.
(4) Adding acetone into the powder, mixing and extracting, wherein the volume mass ratio of the acetone to the powder is 15 ml: 1g, extracting for 40min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(3) And (3) adding distilled water into the extract liquor obtained in the step (2), wherein the adding amount of the distilled water is 3.5 times of the volume of the acetone obtained in the step (2), stirring, standing, filtering and separating out a precipitate after the precipitate is separated out, and thus obtaining a crude product of the vitamin K2 product.
(4) Mixing the crude product of the vitamin K2 with acetone again, wherein the volume mass ratio of the acetone to the crude product is 13 ml: 1g, extracting for 35min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(5) And (4) adding distilled water into the extract liquor obtained in the step (4), wherein the adding amount of the distilled water is 2.5 times of the volume of the acetone obtained in the step (4), stirring, standing, and filtering to separate out a precipitate after the precipitate is separated out, so as to obtain the vitamin K2 product.
(6) And (5) combining the residual extraction liquid after separating the precipitate in the steps (3) and (5), distilling at 60 ℃, and recovering the acetone.
Example 3
A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating the flavobacterium strain to a culture medium in a fermentation tank after seed culture, wherein the culture medium comprises the following components: 7g/L of glycerol, 25g/L of peptone and 3g/L, K of yeast powder2HPO4 5g/L、NaCl 3g/L、MgSO41g/L and 20g/L of agar, wherein the inoculation amount of the flavobacterium strain is 4 percent, the culture temperature is 28 ℃, the stirring speed is 250r/min, and the culture time is 6 days.
(2) And after the culture is finished, adding polyferric chloride into the fermentation liquor to settle the thalli, wherein the concentration of the sedimentation accelerator in the fermentation liquor is 55mg/L, and filtering and separating the thalli for later use after the thalli are completely settled.
(3) And (3) placing the thalli in a freeze dryer, rapidly cooling to-30 ℃, then vacuumizing to 0.2mbra and starting heating, obtaining freeze-dried thalli after water is sublimated, and grinding the freeze-dried thalli into powder.
(4) Adding acetone into the powder, mixing and extracting, wherein the volume mass ratio of the acetone to the powder is 10 ml: 1g, extracting for 30min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(3) And (3) adding distilled water into the extract liquor obtained in the step (2), wherein the adding amount of the distilled water is 3 times of the volume of the acetone obtained in the step (2), stirring, standing, filtering and separating out a precipitate after the precipitate is separated out, and thus obtaining a crude product of the vitamin K2 product.
(4) Mixing the crude product of the vitamin K2 with acetone again, wherein the volume mass ratio of the acetone to the crude product is 8 ml: 1g, extracting for 30min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(5) And (4) adding distilled water into the extract liquor obtained in the step (4), wherein the adding amount of the distilled water is 2.5 times of the volume of the acetone obtained in the step (4), stirring, standing, and filtering to separate out a precipitate after the precipitate is separated out, so as to obtain the vitamin K2 product.
(6) And (5) combining the residual extraction liquid after separating the precipitate in the steps (3) and (5), distilling at 60 ℃, and recovering the acetone.
Example 4
A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating the flavobacterium strain to a culture medium in a fermentation tank after seed culture, wherein the culture medium comprises the following components: 7g/L of glycerol, 25g/L of peptone and 3g/L, K of yeast powder2HPO4 5g/L、NaCl 3g/L、MgSO41g/L and 20g/L of agar, wherein the inoculation amount of the flavobacterium strain is 6 percent, the culture temperature is 25 ℃, the stirring speed is 250r/min, and the culture time is 6 days.
(2) After the culture is finished, alum is added into the fermentation liquor to enable the thalli to settle, the concentration of the sedimentation promoting agent in the fermentation liquor is 40mg/L, and after the thalli are completely settled, the thalli are filtered and separated for later use.
(3) Freezing the thalli obtained in the step (2) at a low temperature of minus 25 ℃ for 5 minutes, and then thawing the thalli at room temperature; repeating the steps for 5 times to ensure that the cells of the thalli are repeatedly frozen, thawed and crushed to obtain the crushed thalli.
(4) Adding acetone into the crushed thallus, mixing and extracting, wherein the volume mass ratio of the acetone to the powder is 12 ml: 1g, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to remove after the extraction is finished, so that the extraction liquid is obtained.
(3) And (3) adding distilled water into the extract liquor obtained in the step (2), wherein the adding amount of the distilled water is 3 times of the volume of the acetone obtained in the step (2), stirring, standing, filtering and separating out a precipitate after the precipitate is separated out, and thus obtaining a crude product of the vitamin K2 product.
(4) Mixing the crude product of the vitamin K2 with acetone again, wherein the volume mass ratio of the acetone to the crude product is 13 ml: 1g, extracting for 35min, continuously stirring in the extraction process, and filtering to remove solid matters to obtain an extract.
(5) And (4) adding distilled water into the extract liquor obtained in the step (4), wherein the adding amount of the distilled water is 2 times of the volume of the acetone obtained in the step (4), stirring, standing, and filtering to separate out a precipitate after the precipitate is separated out, so as to obtain the vitamin K2 product.
(6) The remaining extracts after the precipitates were separated in steps (3) and (5) were combined and then distilled at 65 ℃ to recover acetone.
Example 5
A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating the flavobacterium strain to a culture medium in a fermentation tank after seed culture, wherein the culture medium comprises the following components: 7g/L of glycerol, 25g/L of peptone and 3g/L, K of yeast powder2HPO4 5g/L、NaCl 3g/L、MgSO41g/L and 20g/L of agar, wherein the inoculation amount of the flavobacterium strain is 6 percent, the culture temperature is 25 ℃, the stirring speed is 250r/min, and the culture time is 6 days.
(2) After the culture is finished, alum is added into the fermentation liquor to enable the thalli to settle, the concentration of the sedimentation promoting agent in the fermentation liquor is 40mg/L, and after the thalli are completely settled, the thalli are filtered and separated for later use.
(3) And (3) placing the thalli in a freeze dryer, rapidly cooling to minus 25 ℃, then vacuumizing to 0.1mbra and starting heating, obtaining freeze-dried thalli after water is sublimated, and grinding the freeze-dried thalli into powder.
(4) Adding acetone into the powder, mixing and extracting, wherein the volume mass ratio of the acetone to the powder is 12 ml: 1g, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to remove after the extraction is finished, so that the extraction liquid is obtained.
(3) And (3) adding distilled water into the extract liquor obtained in the step (2), wherein the adding amount of the distilled water is 3 times of the volume of the acetone obtained in the step (2), stirring, standing, filtering and separating out a precipitate after the precipitate is separated out, and thus obtaining a crude product of the vitamin K2 product.
(6) And (4) combining the residual extraction liquid after the precipitate is separated out in the step (3), then distilling at 65 ℃, and recovering the acetone.
The results of testing the purity of vitamin K2 in the vitamin K2 products obtained in examples 1 to 4 and the crude vitamin K2 product prepared in example 5 are shown in Table 1.
TABLE 1
Example 1 Example 2 Example 3 Example 4 Example 5
Vitamin K2 purity/%) 83.9 86.3 84.7 78.1 62.6
From the above detection results, it can be seen that the vitamin K2 in the vitamin K2 products prepared in examples 1 to 3 has high purity, whereas the vitamin K2 in example 5 has low purity because no purification is performed again, and the vitamin K2 in the obtained products is lower than that in examples 1 to 3 because the extraction method is adopted in example 4 after the cell wall is broken by repeated freeze thawing; in addition, this method requires repeated freezing and thawing, and causes loss of the product due to partial product efflux from the disrupted microbial cells during thawing. The method provided by the invention directly prepares the thalli into freeze-dried powder and then extracts the thalli, so that the problem of product loss is solved, and the method is more convenient and efficient.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A method for extracting vitamin K2 from fermentation liquor comprises the following steps:
(1) inoculating flavobacterium to a culture medium for fermentation, adding a sedimentation promoting agent into fermentation liquor after the fermentation is finished, and separating out thalli for later use after the thalli in the fermentation liquor are settled;
(2) freeze-drying the thalli, crushing the thalli into powder, mixing the powder with acetone, extracting, and taking an extract for later use;
(3) adding clear water into the extract, and separating out precipitate after the precipitate is separated out to obtain a crude product of the vitamin K2 product;
(4) and mixing the vitamin K2 product crude product with acetone again for extraction, then adding clear water, and separating out precipitate after the precipitate is separated out to obtain the vitamin K2 product.
2. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein the step (1) comprisesThe following components of culture medium are used: 6-8.5 g/L of glycerol, 20-28 g/L of peptone and 2-3.5 g/L, K of yeast powder2HPO4 3～7.6g/L、NaCl 3～5g/L、MgSO40.1-4 g/L and 15-25 g/L of agar.
3. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in step (1), the inoculum size of Flavobacterium is 4-6.5%, the culture temperature is 24-28 ℃, the stirring speed is 250-350 r/min, and the culture time is 5-6 days.
4. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in step (1), the precipitation promoter comprises any one of aluminum sulfate, ferric sulfate, aluminum chloride, ferric chloride, alum, polyaluminum chloride and polyferric chloride, and preferably, the concentration of the precipitation promoter in the fermentation broth is 40-60 mg/L.
5. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in step (2), the thallus is rapidly cooled to below-20 ℃, then is vacuumized to 0.1-0.2 mbra and is heated, and freeze-dried thallus is obtained after water sublimation.
6. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in the step (2), the volume-to-mass ratio of acetone to powder is 10-15 ml: 1g, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to remove after the extraction is finished, so that the extraction liquid is obtained.
7. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in step (4), the mass-to-volume ratio of the crude vitamin K2 product to acetone is 1 g: 8-13 ml, the extraction time is not less than 30min, stirring is continuously carried out in the extraction process, and solid matters are filtered to obtain the extraction liquid.
8. The method for extracting vitamin K2 from fermentation broth according to claim 1, wherein in steps (3) and (4), the addition amount of the clear water is more than 2 times, preferably 2-3.5 times of the volume of acetone.
9. The method for extracting vitamin K2 from fermentation broth according to claim 8, wherein the clear water comprises any one of distilled water and purified water.
10. The method for extracting vitamin K2 from fermentation broth according to any one of claims 1 to 9, further comprising the step of distilling the residual extract after separating the precipitate to recover acetone therein, wherein the distillation temperature is higher than the boiling point of acetone and lower than the boiling point of water, preferably 60-65 ℃.
CN202110689781.XA 2021-06-22 2021-06-22 Method for extracting vitamin K2 from fermentation liquor Pending CN113308500A (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61216696A (en) * 1985-03-19 1986-09-26 Mitsubishi Gas Chem Co Inc Production of menaquinone-4
JP2004222676A (en) * 2003-01-27 2004-08-12 Natokin:Kk Method for culturing bacillus natto and method for producing viscous material
CN103571897A (en) * 2013-10-29 2014-02-12 中国科学院合肥物质科学研究院 Vitamin K2 and preparation process thereof
CN104673849A (en) * 2015-03-26 2015-06-03 中国科学院合肥物质科学研究院 Process for producing vitamin K2 based on flavobacterium

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61216696A (en) * 1985-03-19 1986-09-26 Mitsubishi Gas Chem Co Inc Production of menaquinone-4
JP2004222676A (en) * 2003-01-27 2004-08-12 Natokin:Kk Method for culturing bacillus natto and method for producing viscous material
CN103571897A (en) * 2013-10-29 2014-02-12 中国科学院合肥物质科学研究院 Vitamin K2 and preparation process thereof
CN104673849A (en) * 2015-03-26 2015-06-03 中国科学院合肥物质科学研究院 Process for producing vitamin K2 based on flavobacterium

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HONGFEIWEI等: "Extraction,purification and identification of menaquinones from Flavobacterium meningosepticum fermentation medium", 《PROCESS BIOCHEMISTRY》 *
朱进伟等: "维生素K2的相关合成研究及前景展望", 《中国抗生素杂志》 *

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Application publication date: 20210827