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Method for producing hyaluronic acid by fermentation and application of method

A technology of hyaluronic acid and fermented liquid, applied in the field of fermentation, can solve the problems of affecting the quality of hyaluronic acid, high production cost, low molecular weight of hyaluronic acid, etc., so as to be beneficial to growth and metabolism, avoid the risk of bacterial contamination, The effect of shortening the fermentation period

Pending Publication Date: 2020-05-19
JIANGSU CHENGXIN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When HA is extracted from animal organs by extraction method, due to limited raw materials and high production cost, the commodity is expensive, which limits its wide application in medicine and cosmetics
Compared with the animal tissue extraction method, the bacterial fermentation method has a production scale that is not limited by animal raw materials. The HA in the fermentation broth exists in a free form, which is easy to separate and purify, with low cost, easy to form large-scale industrial production, and no animal-derived pathogenic virus pollution. In recent years, with the perfection of fermentation production HA technology, the fermentation method has a tendency to completely replace the extraction method, but due to the imbalance between the bacteria's own metabolism and product expression, the production of hyaluronic acid is seriously restricted. and molecular weight
[0004] Disclosed in CN101935678A is a method for producing hyaluronic acid fermentation liquid. In the method, a trace element liquid is added to the culture medium, and the trace element liquid is composed of calcium chloride, zinc chloride, manganese sulfate, and copper sulfate. The preparation of seed medium and fermentation medium with water for injection solves the problem of hyaluronic acid production and quality decline caused by lack of nutrients or improper allocation of various nutrients in the process of bacterial fermentation to produce hyaluronic acid. , the output of hyaluronic acid in this invention is 6-7g / L, and the output is still low
In this invention, the carbon source is supplemented by feeding batches during fermentation, and the carbon source is added five times at different times, so that the yield of hyaluronic acid is relatively high, but the disadvantage of this method is that the carbon source is added many times, and the fermentation The cycle is longer, and it is easy to cause the fermentation broth to be contaminated by bacteria, which affects the quality of hyaluronic acid, and the molecular weight of the obtained hyaluronic acid is low

Method used

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  • Method for producing hyaluronic acid by fermentation and application of method

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Embodiment 1

[0049] This embodiment provides a method for producing hyaluronic acid by fermentation, comprising the following steps:

[0050] 1. Using Streptococcus zooepidemicus as the fermentation strain, inoculate Streptococcus zooepidemicus on the seed solid medium for cultivation, and cultivate at 33°C for 16 hours;

[0051] 2. Inoculate the Streptococcus zooepidemicus into the shake flask seed medium for culture, shake and culture at 33°C for 12 hours, and obtain the Streptococcus zooepidemicus shake flask seed culture solution;

[0052] 3. Inoculate the shake flask seed culture solution of Streptococcus zooepidemicus into the seed tank culture medium for cultivation, the cultivation temperature is 33°C, adjust the pH of the culture solution with 30% sodium hydroxide to maintain it at pH 7.5, and cultivate for 12 hours to obtain Streptococcus zooepidemicus Seed tank seed culture solution;

[0053] 4. Inoculate the seed culture solution of the Streptococcus zooepidemicus seed tank in...

Embodiment 2

[0056] Same as Example 1, the difference is that, in the 8th hour after being transferred to the fermentation medium, 10 g of glucose was supplemented per liter of fermentation broth; the yield of the obtained hyaluronic acid was 10.6 g / L, and the molecular weight of the hyaluronic acid was 2400KDa.

Embodiment 3

[0058] Same as Example 1, the difference is that, at the 12th hour after being transferred to the fermentation medium, 10 g of glucose was supplemented per liter of fermentation broth; the yield of the obtained hyaluronic acid was 10.7 g / L, and the molecular weight of the hyaluronic acid was 3000KDa.

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Abstract

The invention provides a method for producing hyaluronic acid by fermentation and an application of the method. The method comprises the following steps: performing activation and propagation on streptococcus zooepidemicus, then performing fermentation, supplementing a carbon source for once in the fermentation process to obtain a hyaluronic acid fermentation broth, and performing purifying to obtain hyaluronic acid. According to the fermentation method provided by the invention, the carbon source is added for once in the fermentation process, so that the glucose concentration in the fermentation broth can be maintained, the fermentation period is shortened, the contamination risk caused by adding the carbon source for a plurality of times is avoided, and normal growth and metabolism of thalli are facilitated. The invention solves the problem of unbalance between metabolism of streptococcus zooepidemicus and expression of products is solved, and the obtained hyaluronic acid has a highyield and a large molecular weight.

Description

technical field [0001] The invention belongs to the technical field of fermentation, and in particular relates to a method for fermenting and producing hyaluronic acid, in particular to a method for fermenting and producing polymer pharmaceutical grade hyaluronic acid and its application. Background technique [0002] Hyaluronic acid (HyaluroIlic acid, HA for short), also known as hyaluronic acid, is a macromolecular mucopolysaccharide composed of glucuronic acid and N-acetylglucosamine repeatedly alternating through β-1,3 and β-1,4 glycosidic bonds. Linked into a chain polymer. Based on hyaluronic acid's high viscosity, lubricity, water retention, good biocompatibility and special physiological effects, hyaluronic acid has a wide range of applications in medicine, beauty, health food and other fields. Different uses of HA have different requirements on molecular weight. HA with medium molecular weight (molecular weight range 1,000-1,800KDa) is mainly used in cosmetics, whi...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12N1/20C12R1/46
CPCC12P19/26C12N1/20
Inventor 李斌徐飞祝俊余允东马晶晶
Owner JIANGSU CHENGXIN PHARMA
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