A kind of method for improving hyaluronic acid yield

A technology of hyaluronic acid and hyaluronidase, applied in the field of bioengineering, can solve the problems of prolonging the fermentation time and decreasing the yield of hyaluronic acid, and achieve the effects of improving mass transfer efficiency, increasing biosynthesis rate, and increasing yield

Active Publication Date: 2022-08-09
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the above-mentioned problems existing in the prior art, the present invention provides a method for increasing the yield of hyaluronic acid, thereby overcoming the problem that the yield of hyaluronic acid decreases due to early addition of enzymes, and prolonging the fermentation time

Method used

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  • A kind of method for improving hyaluronic acid yield
  • A kind of method for improving hyaluronic acid yield

Examples

Experimental program
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Effect test

Embodiment 1

[0046] The strain of Corynebacterium glutamicum was inoculated into a 250mL shake flask containing 50mL activation medium (brain heart infusion medium 37g / L), and cultured for 10-12h.

[0047] Incubate at 30°C for 15h, and inoculate 50mL of the above-mentioned culture medium in a 2.5L fermenter for cultivation. Medium: yeast extract 20g / L, peptone 3g / L, MgSO 4 1.2g / L, glucose 40g / L, PTM1 1mL / L, 5mL / L Tween 80.

[0048] 24h before fermentation, the dissolved oxygen was linked with the rotation speed to keep the dissolved oxygen not less than 10%, and 600U / mL of leech hyaluronidase was added at the 8th hour. In the subsequent fermentation, as the content of hyaluronic acid in the fermentation broth increases, the viscosity of the fermentation broth gradually increases, and the dissolved oxygen gradually decreases. , add 600U / mL of leech hyaluronidase, specifically to this example, add 10 times in total. At 72h, the content of hyaluronic acid in the fermentation broth was 59....

Embodiment 2

[0050] The strain of Corynebacterium glutamicum was inoculated into a 250mL shake flask containing 50mL activation medium (brain heart infusion medium 37g / L), and cultured for 10-12h.

[0051] Incubate at 30°C for 15h, inoculate 50mL of the above culture solution into a 2.5L fermenter for culture medium: yeast extract 20g / L, peptone 3g / L, MgSO 4 1.2g / L, glucose 40g / L, PTM1 1mL / L, 10mL / L Tween 80.

[0052] 24h before fermentation, the dissolved oxygen was linked with the rotation speed to keep the dissolved oxygen not less than 1%, and 600U / mL of bacterial hyaluronan lyase was added at the 8th hour. In the subsequent fermentation, with the increase of the hyaluronic acid content in the fermentation broth, the viscosity of the fermentation broth gradually increased, and the dissolved oxygen gradually decreased. , adding 600U / mL of bacterial hyaluronan lyase, specifically to this example, adding a total of 10 times. At 72h, the content of hyaluronic acid in the fermentation br...

Embodiment 3

[0054] Inoculate Streptococcus zooepidemicus into a 250mL shake flask containing 50mL activation medium (brain heart infusion medium 37g / L), and cultivate for 10-12h.

[0055] Incubate at 30°C for 15h, inoculate 50mL of the above culture solution into a 2.5L fermenter for culture medium: yeast extract 20g / L, peptone 3g / L, MgSO 4 1.2g / L, glucose 40g / L, PTM1 1mL / L, 5mL / L Tween 80.

[0056] 24h before fermentation, the dissolved oxygen was linked with the rotation speed to keep the dissolved oxygen not less than 3%, and 600U / mL of leech hyaluronidase was added at the 8th hour. In the subsequent fermentation, with the increase of hyaluronic acid content in the fermentation broth, the viscosity of the fermentation broth gradually increased, and the dissolved oxygen gradually decreased. , add 600U / mL of leech hyaluronidase, a total of 10 times. After 72h of culture, the content of HA in the fermentation broth was 27.3g / L, and the molecular weight was 10kDa-40kDa.

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Abstract

The present invention disclosed a method that improves the production rate of hyaluronic acid. The method is added in the fermentation solution of hyaluronic acid to add hyaluronase according to the soluble oxygen level of the fermentation solution. The fermentation solution is in the fermentation mediumThe bacteria of synthetic hyaluronic acid are obtained for fermentation.The method of the present invention can realize a one -step legal system preparing small molecular hyaluronic acid. This method can be applied to any strain that fermented synthetic hyaluronic acid, and any enzyme that can degrade hyaluronic acid.In addition, the present invention can protect the cells to a certain extent by adding a vomiting temperature 80 to solve the adverse effects of the early hyaluronase on the bacteria, and reduce the damage of the cells on cells.And adding hyaluronase in fermentation can reduce the viscosity of the fermentation liquid, improve the efficiency of the mass transmission, and help improve the bio -synthesis rate of early hyaluronic acid, thereby increasing the output of hyaluronic acid.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for improving the yield of hyaluronic acid. Background technique [0002] Hyaluronic acid (HA), commonly known as hyaluronic acid, is a repeating disaccharide unit composed of D-glucuronic acid and N-acetylglucosamine, through β(1-3) and β(1-4) glycosides. The macromolecular viscous polysaccharide, which is alternately connected by bonds, was first extracted from bovine glass spheres by Meyer et al. in 1934. With its unique molecular structure and physicochemical properties, hyaluronic acid shows a variety of important physiological functions in the body, such as lubricating joints, regulating the permeability of blood vessel walls, regulating proteins, water and electrolyte diffusion and operation, and promoting wound healing. It is particularly important that hyaluronic acid has a special water-retaining effect and is the substance with the best moi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12R1/46C12R1/125C12R1/15C12R1/19
CPCC12P19/26
Inventor 张天萌戚明刘金钊罗德学刘英杰张由恒王景郝井坤
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD
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