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Recombinant human hyaluronic acid and production method thereof

A technology of human hyaluronidase and hyaluronic acid, which is applied in the field of daily chemical industry to achieve the effects of increasing production, reducing inflammation and promoting the production of IL-10

Inactive Publication Date: 2018-05-04
杭州惠博士生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The study found that the method of using general hyaluronidase to degrade macromolecular hyaluronic acid to obtain small molecular hyaluronic acid fragments is affected by the homology of the enzyme, the half-life of the enzyme, the source of raw materials, and the enzymatic hydrolysis process, which affect the product yield and human activity of the product. However, it needs to be further optimized and solved urgently, so it is imminent to find a more effective method for preparing small molecule hyaluronic acid with human activity

Method used

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  • Recombinant human hyaluronic acid and production method thereof
  • Recombinant human hyaluronic acid and production method thereof
  • Recombinant human hyaluronic acid and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] ① Take some 250ml Erlenmeyer flasks, add 100ml of purified water respectively, add NaCl to each Erlenmeyer flask after marking to make the concentration 0.5mol / L, and adjust the pH to 7.0;

[0047] ②Add 3g macromolecular hyaluronic acid dry powder to each triangular flask to make the concentration 30g / L, dissolve overnight;

[0048] ③After the macromolecular hyaluronic acid is completely dissolved, add recombinant human hyaluronidase solution to each conical flask, and control the enzyme activity concentration at 400U / L;

[0049] ④Stir evenly at 1500r / min, place in a constant temperature shaker, digest at 40°C for 4 hours, and control the pH to 5.0-5.6 during this period;

[0050] ⑤Wait until the enzyme digestion is finished, increase the temperature of the enzymatic hydrolysis solution to 80°C, and inactivate the enzyme for 30 minutes;

[0051] ⑥Add 300ml of 95% ethanol solution to each sample for precipitation, discard the supernatant after precipitation, dehydrate w...

Embodiment 2

[0054] ① Take some 250ml Erlenmeyer flasks, add 100ml of purified water respectively, add NaCl to each Erlenmeyer flask after marking to make the concentration 0.5mol / L, and adjust the pH to 7.0;

[0055] ②Add 3g macromolecular hyaluronic acid dry powder to each triangular flask to make the concentration 30g / L, dissolve overnight;

[0056] ③After the macromolecular hyaluronic acid is completely dissolved, add recombinant human hyaluronidase solution to each conical flask, and control the enzyme activity concentration at 600U / L;

[0057] ④Stir evenly at 1500r / min, place in a constant temperature shaker, digest at 40°C for 4 hours, and control the pH to 5.0-5.6 during this period;

[0058] ⑤Wait until the enzyme digestion is finished, increase the temperature of the enzymatic hydrolysis solution to 80°C, and inactivate the enzyme for 30 minutes;

[0059] ⑥Add 300ml of 95% ethanol solution to each sample for precipitation, discard the supernatant after precipitation, dehydrate w...

Embodiment 3

[0062] ① Take some 250ml Erlenmeyer flasks, add 100ml of purified water respectively, add NaCl to each Erlenmeyer flask after marking to make the concentration 0.5mol / L, and adjust the pH to 7.0;

[0063] ②Add 3g macromolecular hyaluronic acid dry powder to each triangular flask to make the concentration 30g / L, dissolve overnight;

[0064] ③After the macromolecular hyaluronic acid is completely dissolved, add recombinant human hyaluronidase solution to each conical flask, and control the enzyme activity concentration at 1000U / L;

[0065] ④Stir evenly at 1500r / min, place in a constant temperature shaker, digest at 40°C for 4 hours, and control the pH to 5.0-5.6 during this period;

[0066] ⑤Wait until the enzyme digestion is finished, increase the temperature of the enzymatic hydrolysis solution to 80°C, and inactivate the enzyme for 30 minutes;

[0067] ⑥Add 300ml of 95% ethanol solution to each sample for precipitation, discard the supernatant after precipitation, dehydrate ...

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Abstract

The invention provides recombinant human hyaluronic acid which is homologous with the human body. The structural formula of the recombinant human hyaluronic acid is as follows: a formula is as shown in the description, wherein n is 3-10. Two ends of the structural formula are enzyme digestion tail ends which are homologous with the human body. The recombinant human hyaluronic acid prepared by themethod also has functions of inhibiting bacteria and diminishing inflammation and the improvement of the human activity of enhancing skin mucous membrane immunity besides the functions of ordinary hyaluronic acid. The recombinant human hyaluronic acid can be widely applied to beautifying and conditioning of the aspects such as face, oral cavity, hair, body and private parts of the human body, andcan be prepared into a daily chemical product with higher safety coefficient, more durable stability and higher practicability.

Description

technical field [0001] The invention relates to the field of daily chemical industry, in particular to a recombinant human hyaluronic acid and a production method thereof. Background technique [0002] Hyaluronic acid, also known as hyaluronic acid, is a high-molecular polymer. It is an advanced acidic mucopolysaccharide composed of units of D-glucuronic acid and N-acetylglucosamine. It is an important basic substance for skin suppleness. It is also a component of the human body. It has a special water-retaining effect, and its weight is as high as 100 times its own weight. It is the most moisturizing substance found in nature. It can also improve skin nutrition metabolism and make skin soft and smooth. , anti-wrinkle, increase elasticity, prevent aging, while moisturizing, it is also a good transdermal absorption accelerator, used in conjunction with other nutrients, can achieve a more ideal effect of promoting nutrient absorption. Among the existing sources of hyaluronic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/26C12N15/85C12P19/26C12P19/14
CPCC12N9/2474C12N15/85C12P19/14C12P19/26C12Y302/01035
Inventor 贺晓军
Owner 杭州惠博士生物科技有限公司
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