Method for preparing small-molecular weight hyaluronic acid by adding hydrogen phosphide and ascorbic acid in course of fermentation

Abstract

The invention discloses a method for producing hyaluronic acid with small molecular weight by adding hydrogen peroxide and ascorbic acid in fermentation process, which belongs to the technology field of the application of oxidative-reductive degradation reaction in optimization of fermentation process. In the method, hydrogen peroxide (0.1 to 1.0 mmol / g hyaluronic acid) and ascorbic acid (0.05 to 0.5 mmol / g hyaluronic acid) are added in the fermentation process (8 h) to degrade hyaluronic acid, so that the viscosity of a fermentation broth decreases from reference 300 cp (without hyaluronidase) to 100 to 200 cp, the dissolved oxygen concentration is improved from reference 0 to 1% to 5 to 10%, the output of hyaluronic acid is improved from reference 5.0 g / L to 5.5 to 6.4 g / L, and the molecular weight of the hyaluronic acid decreases from reference 1,300 kDa to 80 to 200 kDa. The inventive method is simple and efficient, and has important industrial application value. The decrease of the viscosity of the fermentation broth results in the improvement of downstream purification efficiency and the reduction of production cost. The hyaluronic acid with small molecular weight produced by the method can be applied to the fields of medicines and healthy food.

Description

technical field

[0001] The invention discloses a method for producing low-molecular-weight hyaluronic acid by adding hydrogen peroxide and ascorbic acid in the fermentation process, which belongs to the technical field of redox degradation reaction optimization application in the fermentation process. Background technique

[0002] Hyaluronic acid is a linear anionic mucopolysaccharide formed by glucuronic acid and N-acetylglucosamine linked by β-1, 3 and β-1, 4 glycosidic bonds. It was obtained from bovine vitreous by Meyer and Palmer in 1934 seperate. Hyaluronic acid is an important component of extracellular matrix, joint synovial fluid, cartilage tissue, and umbilical cord. It has important physiological functions in the human body, such as regulating the chemical composition of extracellular fluid, lubricating and promoting wound healing; regulating adhesion, growth, Movement and activation of specific cells, etc.

[0003] The molecular weight of hyaluronic acid is clo...

Examples

Embodiment 1

[0010] Example 1: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (0.1 mmol / g hyaluronic acid) and ascorbic acid (0.05 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 5.5g / L with a molecular weight of 200kDa.

Embodiment 2

[0011] Example 2: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (0.6 mmol / g hyaluronic acid) and ascorbic acid (0.3 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 5.7g / L with a molecular weight of 120kDa.

Embodiment 3

[0012] Example 3: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (1.0 mmol / g hyaluronic acid) and ascorbic acid (0.5 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 6.5g / L with a molecular weight of 80kDa.

technical field

[0001] The invention discloses a method for producing low-molecular-weight hyaluronic acid by adding hydrogen peroxide and ascorbic acid in the fermentation process, which belongs to the technical field of redox degradation reaction optimization application in the fermentation process. Background technique

[0002] Hyaluronic acid is a linear anionic mucopolysaccharide formed by glucuronic acid and N-acetylglucosamine linked by β-1, 3 and β-1, 4 glycosidic bonds. It was obtained from bovine vitreous by Meyer and Palmer in 1934 seperate. Hyaluronic acid is an important component of extracellular matrix, joint synovial fluid, cartilage tissue, and umbilical cord. It has important physiological functions in the human body, such as regulating the chemical composition of extracellular fluid, lubricating and promoting wound healing; regulating adhesion, growth, Movement and activation of specific cells, etc.

[0003] The molecular weight of hyaluronic acid is clo...

Examples

Embodiment 1

[0010] Example 1: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (0.1 mmol / g hyaluronic acid) and ascorbic acid (0.05 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 5.5g / L with a molecular weight of 200kDa.

Embodiment 2

[0011] Example 2: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (0.6 mmol / g hyaluronic acid) and ascorbic acid (0.3 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 5.7g / L with a molecular weight of 120kDa.

Embodiment 3

[0012] Example 3: Fermentation in a fermenter adopts the strategy of adding hydrogen peroxide and ascorbic acid, and the rest of the conditions are the same as those in the comparative example. Hydrogen peroxide (1.0 mmol / g hyaluronic acid) and ascorbic acid (0.5 mmol / g hyaluronic acid) were added at 8 h of fermentation. At the end of the fermentation, the production of hyaluronic acid increased to 6.5g / L with a molecular weight of 80kDa.