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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, etc., and achieve the goal of improving mass transfer efficiency, biosynthesis rate, and mass transfer efficiency Effect

Active Publication Date: 2021-03-26
BLOOMAGE BIOTECHNOLOGY CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Method for improving hyaluronic acid yield

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Corynebacterium glutamicum strains were inoculated into 250mL shake flasks containing 50mL activation medium (brain heart infusion medium 37g / L), and cultivated for 10-12h.

[0047] Cultivate at 30° C. for 15 hours, inoculate 50 mL of the above culture solution into a 2.5 L 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] 24 hours before fermentation, the dissolved oxygen was linked with the rotational speed to keep the dissolved oxygen not lower than 10%, and 600 U / mL of leech hyaluronidase was added at the 8th hour. In the subsequent fermentation, as the hyaluronic acid content in the fermentation broth increases, the viscosity of the fermentation broth increases gradually, and the dissolved oxygen gradually decreases. The dissolved oxygen is coupled with the hyaluronidase, and whenever the dissolved oxygen is lower than 10%, , adding 600 U / mL of leech hyaluronidase, specific to...

Embodiment 2

[0050] Corynebacterium glutamicum strains were inoculated into 250mL shake flasks containing 50mL activation medium (brain heart infusion medium 37g / L), and cultivated for 10-12h.

[0051] Cultivate at 30°C for 15 hours, 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] 24 hours before fermentation, the dissolved oxygen was linked with the rotational speed to keep the dissolved oxygen not lower than 1%, and 600 U / mL of bacterial hyaluronan lyase was added at the 8th hour. In the subsequent fermentation, as the hyaluronic acid content in the fermentation broth increases, the viscosity of the fermentation broth increases gradually, and the dissolved oxygen gradually decreases. The dissolved oxygen is coupled with the hyaluronidase, and whenever the dissolved oxygen is lower than 1%, , adding 600 U / mL bacterial hyaluronan lyase, specific to...

Embodiment 3

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

[0055] Cultivate at 30°C for 15 hours, 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] 24 hours before fermentation, the dissolved oxygen was linked with the rotational speed to keep the dissolved oxygen not lower than 3%, and 600 U / 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 increases gradually, and the dissolved oxygen gradually decreases. The dissolved oxygen is coupled with the hyaluronidase, and whenever the dissolved oxygen is lower than 3% , add 600U / mL leech hyaluronidase, and add 10 times in total. After cul...

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Abstract

The invention discloses a method for improving hyaluronic acid yield. The method is characterized in that hyaluronidase is added into fermentation liquor for producing hyaluronic acid according to thedissolved oxygen level of the fermentation liquor, and the fermentation liquor is obtained through fermentation by adding bacteria for synthesizing hyaluronic acid into a fermentation culture medium.The method can realize one-step preparation of small molecular hyaluronic acid, and can be applied to any bacterial strain for fermentation synthesis of hyaluronic acid and any enzyme capable of degrading hyaluronic acid; besides, tween 80 is added, so that cells are protected to a certain extent, the adverse effect of early-stage hyaluronidase on bacterial cells is avoided, and the damage of hyaluronidase to the cells is reduced; and the hyaluronidase is added in the fermentation liquor, so that the viscosity of fermentation liquor can be reduced, the mass transfer efficiency is improved, the biosynthesis rate of hyaluronic acid in the earlier stage can be increased, and the yield of hyaluronic acid is increased.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for increasing the yield of hyaluronic acid. Background technique [0002] Hyaluronic acid (HA for short), commonly known as hyaluronic acid, is a repeating disaccharide unit consisting of D-glucuronic acid and N-acetylglucosamine, through β(1-3) and β(1-4) glycosides It is a macromolecular viscous polysaccharide formed by alternating bonds, which was first extracted from bovine glass globe eyes by Meyer et al. in 1934. With its unique molecular structure and physical and chemical 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 protein, water and electrolyte diffusion and operation, and promoting wound healing. More importantly, hyaluronic acid has a special water-retaining effect and is the most moisturizing substanc...

Claims

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Application Information

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Patent Type & Authority Applications(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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