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A method for producing high molecular weight hyaluronic acid by fermentation

A technology of high molecular weight and hyaluronic acid, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of reducing the added value of products, requiring high cultivation conditions, and not being able to form a single product of purity, etc., to achieve Effects of improving purity and increasing yield

Active Publication Date: 2020-10-09
XINJIANG FUFENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The applicant's previous patented technology CN105368912A uses two strains for mixed fermentation, which greatly improves the fermentation efficiency, but this method requires high culture conditions, which may result in the inability to form dominant strains, and the produced hyaluronic acid includes three types: small, medium and large. Molecular weight hyaluronic acid cannot form a single product with high purity, which reduces the added value of the product

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A method for producing high molecular weight hyaluronic acid by fermentation, comprising the following steps:

[0032] Streptococcus zooepidemicus was taken out and inoculated into an activation medium whose composition was 75 g / L of sucrose, 2.5 g / L of peptone, and 1.5 g / L of magnesium sulfate for activation;

[0033] Inoculate the activated strain into a 5L fermenter for fermentation. The fermentation formula is 70g / L sucrose, 2.0g / L peptone, 1.8g / L magnesium sulfate, 1.5g / L potassium sulfate, pH 7.1, temperature 39.5°C , dissolved oxygen 3.1L / min; fermentation 6h, add glycerin to the fermenter, control the concentration of glycerol to 200mg / L; 3 seconds, the interval time is 3 seconds, and the total time of ultrasonic treatment is 90 seconds; then continue to ferment for 2 hours, and stop the fermentation; the total time of fermentation is 16 hours; during the fermentation process, the residual sugar concentration is controlled by adding a sucrose solution with a con...

Embodiment 2

[0037] A method for producing high molecular weight hyaluronic acid by fermentation, comprising the following steps:

[0038] Take Streptococcus zooepidemicus out and inoculate it into an activation medium whose main components are sucrose 75g / L, peptone 2.5g / L, and magnesium sulfate 1.5g / L for activation;

[0039] Inoculate the activated strain into a 10L fermenter for fermentation. The fermentation formula is 70g / L sucrose, 2.0g / L peptone, 1.8g / L magnesium sulfate, 1.5g / L potassium sulfate, pH 7.0, temperature 39.8°C , dissolved oxygen 3.0L / min;

[0040] Ferment to 7h, add glycerin to the fermenter, the concentration of glycerol is 300mg / L; ferment to 15h, ultrasonic treatment, ultrasonic treatment parameters: ultrasonic power is 300w, ultrasonic action time is 3s each time, interval time is 3s , the total time of ultrasonic treatment is 120s; then continue to ferment for 2h, and stop the fermentation; during the fermentation process, the residual sugar concentration is con...

Embodiment 3

[0044] Effects of various factors in the fermentation process on the yield and molecular weight of hyaluronic acid:

[0045] Affecting factor one: glycerin. Taking Example 1 as an example, a plurality of glycerin concentration groups are set, which are 0mg / L, 100mg / L, 200mg / L, 300mg / L, 400mg / L, 500mg / L, and the rest are the same as Example 1. Detect the yield and molecular weight of hyaluronic acid in the fermentation broth, see Table 1 for details:

[0046] Table 1

[0047] Glycerin concentration mg / L Hyaluronic acid output g / L Molecular weight of hyaluronic acid (10,000 Da) 0 2.53 182 100 2.98 197 200 3.79 239 300 3.64 243 400 3.27 248 500 3.01 251

[0048]Conclusion: As shown in Table 1, the addition of glycerol can significantly improve the results of hyaluronic acid. The fermentation yield of glycerol concentration is the highest at 200mg / L. With the increase of glycerol concentration, the production of hyaluronic aci...

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Abstract

The invention belongs to the field of biological fermentation of hyaluronic acid, and discloses a method for preparing HMW (high molecular weight) hyaluronic acid by adopting a fermentation method. The method comprises the following steps: (1) carrying out strain activation; (2) carrying out fermentation; and (3) carrying out filtering, carrying out alcohol precipitation, carrying out centrifugalizing, and carrying out drying. The method provided by the invention is simple and feasible, and the prepared hyaluronic acid is high in yield and molecular weight, is pure white and transparent in appearance, and achieves the requirements of the cosmetic-level products.

Description

technical field [0001] The invention belongs to the field of hyaluronic acid biological fermentation, in particular to a method for producing high molecular weight hyaluronic acid by fermentation. Background technique [0002] The chemical name of hyaluronic acid (HA) is uronic (glass) acid, which is composed of D-glucuronic acid and N-acetylglucosamine monomers, which are repeatedly and alternately linked by β-1, 4 glycosidic bonds. And the chain polymer acidic mucopolysaccharide formed. HA is usually soluble in water, insoluble in organic solvents such as alcohol, ketone, ether, etc. It has the functions of moisturizing, anti-aging, and lubrication, and is widely used in daily chemical, health food, medicine and other fields; for example, in the field of medicine, HA can be used in medicine As a drug carrier, it can target various drugs to pathological sites, and has the effects of promoting fracture healing and certain anticancer effects. [0003] Sodium hyaluronate pro...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12R1/46
CPCC12P19/26
Inventor 李学朋魏璐包鑫张宗华崔国梁刘元涛杨瑞丽
Owner XINJIANG FUFENG BIOTECH
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