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Biological fermentation extracting method for hyaluronic acid

A technology of biological fermentation and extraction methods, applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions, can solve problems such as large environmental pollution and potential safety hazards, achieve short production cycle, low cost, and ensure natural whiteness degree of effect

Inactive Publication Date: 2010-12-15
湖北远成药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method uses a large amount of organic solvents such as acetone, which causes great environmental pollution and potential safety hazards, while the biological fermentation method is safe, reliable and pollution-free

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A biofermentation extraction method for hyaluronic acid, the steps of which are:

[0045] A. Streptococcus zooepidemicus was used as the original strain, and the strain was mutated by physical mutagen ultraviolet rays and chemical mutagen nitrosoguanidine. Then through the isolation, primary screening, re-screening and performance testing of the mutagenic strains, the production strains with good production performance are obtained;

[0046] B. inoculate the bacterial strain on the slant agar medium, and culture at a constant temperature of 40° C. for 48 hours to obtain the slant strain;

[0047] C. Add water 500ml glucose 0.5%, beef extract 0.5%, peptone 1%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.1% to the 1000ml Erlenmeyer flask according to the ratio of water and raw materials to prepare culture solution, sterilize at 120°C for 30min, cool down to 37°C, put the slant bacteria into the Erlenmeyer flask, and incubate at a constant temperature of 37°C...

Embodiment 2

[0055] A. Streptococcus equi-like was used as the original strain, and the strain was mutated by physical mutagen ultraviolet rays and chemical mutagen nitrosoguanidine. Then through the isolation, primary screening, re-screening and performance testing of the mutagenic strains, the production strains with good production performance are obtained;

[0056] B. inoculate the bacterial strain on the slant agar medium, and culture at a constant temperature of 40° C. for 36 hours to obtain the slant strain;

[0057] C. Add 500ml of water, glucose 0.5%, beef extract 0.5%, peptone 1%, dipotassium hydrogen phosphate 0.2%, and magnesium sulfate 0.1% to a 1000ml Erlenmeyer flask according to the ratio of water and raw materials to prepare a culture solution, sterilize at 120°C for 30min, and cool down to 37°C, put the slant bacteria into the Erlenmeyer flask, and incubate at 37°C for 16 hours;

[0058]D. Access the seed tank according to the ratio of 1:10, expand the liquid strain (the...

Embodiment 3

[0065] A. Streptococcus equi was used as the original strain, and the strain was mutated by physical mutagen ultraviolet rays and chemical mutagen nitrosoguanidine. Then through the isolation, primary screening, re-screening and performance testing of the mutagenic strains, the production strains with good production performance are obtained;

[0066] B. inoculate the bacterial strain on the slant agar medium, and culture at a constant temperature of 40° C. for 36 hours to obtain the slant strain;

[0067] Add water 500ml glucose 0.5%, beef extract 0.5%, peptone 1%, potassium dihydrogen phosphate 0.2%, potassium hydrogen phosphate 0.2%, magnesium sulfate 1% preparation nutrient solution in C, 1000ml conical flask by the ratio of water and raw material, Sterilize at 120°C for 30 minutes, cool down to 37°C, transfer the slanted bacteria into the Erlenmeyer flask, and incubate at 37°C for 15 hours;

[0068] D. Access the seed tank according to the ratio of 1:10, expand the liqui...

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PUM

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Abstract

The invention discloses a biological fermentation extracting method for hyaluronic acid. The method comprises the following steps of: A, taking streptococcus as an original strain, and taking ultraviolet rays and nitrosoguanidine as mutagens; B, inoculating mutagenic strains to an agar culture medium, and culturing slant strains; C, inoculating the slant strains to a triangular flask; D, inoculating the triangular flask strains to a seeding tank according to a proportion to perform spread culture; E, sterilizing and cooling culture solution in a fermentation tank, and inoculating seeds to the fermentation tank to perform fermentation; F, inactivating the fermentation solution with heating; G, adding active carbon into the fermentation solution to filter decarburized bacteria; H, adding CPB aqueous solution into the filtrate with stirring, and standing and settling the solution; I, adding sodium chloride solution of fermentation solution volume into the sediment, and stirring the solution; J, decolorizing dissociation solution by ion exchange; K, concentrating and purifying the decolorized solution by membrane separation; L, adding ethanol into the concentrate, and separating out the sediment; and M, drying the sediment to obtain the hyaluronic acid. The method has the advantages of feasibility, simple and convenient operation, short production period, low cost, high efficiency, energy conservation and environmental protection. The hyaluronic acid can be widely used in the fields of cosmetics, food, medicaments and the like.

Description

technical field [0001] The invention relates to the technical field of biological fermentation (gene, cell, fermentation engineering), and more specifically relates to a method for extracting hyaluronic acid through biological fermentation. Background technique [0002] Hyaluronic acid widely exists in various tissues of animals, and its important functions are to maintain cell moisture, improve wound healing and regeneration ability, reduce scars, and enhance immunity. It can be used for non-steroidal anti-inflammatory drugs, arthritis treatment, auxiliary medicine for ophthalmology and heart surgery. It has a unique effect in treating scalds, burns, frostbite, and artificial skin. At present, the production of hyaluronic acid is extracted from animal tissues such as chickens and sheep, which has high restrictions on raw materials and high production costs. The extraction of hyaluronic acid by biological fermentation is not limited by raw materials, and is safe and reliabl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C12N13/00C12N15/01C08B37/08C12R1/465
CPCY02P20/10
Inventor 张晓东叶传发
Owner 湖北远成药业有限公司
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