Method for immobilizing microorganism high-yield hyaluronic acid by using 3D printing technology

A 3D printing and hyaluronic acid technology, applied in the field of hyaluronic acid preparation, can solve the problems of high operating cost, low reusability, and complicated purification steps, and achieve simplified purification steps, high antioxidant activity, and short production cycle. Effect

Pending Publication Date: 2022-05-06
SUN YAT SEN UNIV SHENZHEN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the yield of hyaluronic acid produced by microbial fermentation is generally not high, and there are disadvantages such as high operating cost, low reusability, cumbersome purification steps, and inability to carry out l...

Method used

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  • Method for immobilizing microorganism high-yield hyaluronic acid by using 3D printing technology
  • Method for immobilizing microorganism high-yield hyaluronic acid by using 3D printing technology
  • Method for immobilizing microorganism high-yield hyaluronic acid by using 3D printing technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Preparation method of high-yield hyaluronic acid by immobilized microorganism using 3D printing technology

[0040] (1) A-type pigskin gelatin was dissolved in phosphate buffer (pH 7.4) at 50℃ to prepare 10% gelatin solution, and then methacrylic anhydride (0.6g of methacrylic anhydride per 1g of gelatin) was dripped into the gelatin solution at 50℃, stirred and reacted for 3h. After 3 hours of reaction, the solution was centrifuged at 3500g for 3min, the supernatant was collected and heated at four times the volume. The diluted gelatin solution was dialyzed with deionized water (MWCO, 8000-14000 DA) at 40℃ for 7 days, and the product was collected by freeze-drying to obtain the modified GelMA.

[0041](2) Weigh 0.25g modified GelMA and 0.25g gel (type A pigskin gelatin) in 4.5mL deionized water, stir and mix the obtained mixture at a constant speed of 500rpm at 37℃, and after it is completely dissolved, mix the mixture with LAP (the mass-volume ratio to the deion...

Embodiment 2

[0046] Example 2 Preparation method of high-yield hyaluronic acid by immobilized microorganism using 3D printing technology

[0047] (1) A-type pigskin gelatin was dissolved in phosphate buffer (pH 7.4) at 50℃ to prepare 10% gelatin solution, and then methacrylic anhydride (0.6g of methacrylic anhydride per 1g of gelatin) was dripped into the gelatin solution at 50℃, stirred and reacted for 3h. After 3 hours of reaction, the solution was centrifuged at 3500g for 3min, the supernatant was collected and heated at four times the volume. The diluted gelatin solution was dialyzed with deionized water (MWCO, 8000-14000 DA) at 40℃ for 7 days, and the product was collected by freeze-drying to obtain the modified GelMA.

[0048] (2) Weigh 0.25g modified GelMA and 0.25g gel (type A pigskin gelatin) in 4.5mL deionized water, stir and mix the obtained mixture at a constant speed of 500rpm at 37℃, and after it is completely dissolved, mix the mixture with LAP (the mass-volume ratio to the deio...

Embodiment 3

[0052] Example 3 Preparation method of high-yield hyaluronic acid by immobilized microorganism using 3D printing technology

[0053] (1) A-type pigskin gelatin was dissolved in phosphate buffer (pH 7.4) at 50℃ to prepare 10% gelatin solution, and then methacrylic anhydride (0.6g of methacrylic anhydride per 1g of gelatin) was dripped into the gelatin solution at 50℃, stirred and reacted for 3h. After 3 hours of reaction, the solution was centrifuged at 3500g for 3min, the supernatant was collected and heated at four times the volume. The diluted gelatin solution was dialyzed with deionized water (MWCO, 8000-14000 DA) at 40℃ for 7 days, and the product was collected by freeze-drying to obtain the modified GelMA.

[0054] (2) Weigh 0.25g modified GelMA and 0.25g gel (type A pigskin gelatin) in 4.5mL deionized water, stir and mix the obtained mixture at a constant speed of 500rpm at 37℃, and after it is completely dissolved, mix the mixture with LAP (the mass-volume ratio to the deio...

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Abstract

The invention belongs to the technical field of hyaluronic acid preparation, and particularly relates to a method for high-yield hyaluronic acid by immobilizing microorganisms through a 3D printing technology, GelMA is prepared through a reaction of gelatin and a methacrylic acid esterification reagent, then Gel, GelMA and a photoinitiator are mixed to prepare bio-ink based on Gel-GelMA, and the bio-ink is applied to preparation of high-yield hyaluronic acid. Then, streptococcus equi subspecies for producing hyaluronic acid are uniformly loaded into bio-ink based on Gel-GelMA, then, an immobilized reactor loaded with microorganisms is prepared through a 3D printing technology, and finally, hyaluronic acid is produced in fermentation liquor containing carbon source and nitrogen source substances. The method provided by the invention can solve the defects of low yield, high cost and the like of the current planktonic microorganism direct fermentation method, improves the yield of hyaluronic acid, and can easily separate hyaluronic acid from surrounding media by utilizing a structure containing immobilized bacteria, thereby simplifying the acquisition process of hyaluronic acid.

Description

technical field [0001] The invention belongs to the technical field of hyaluronic acid preparation, and in particular relates to a method for immobilizing microorganisms with high yield of hyaluronic acid by using 3D printing technology. Background technique [0002] Hyaluronic acid is a viscoelastic and hygroscopic glycosaminoglycan polymer composed of repeated glucuronic acid and N-acetyl glucosaminoglycans through β-1,4 glycosidic bonds, widely present in various parts of the human body and in the intercellular space. As a natural polymer material from organisms, hyaluronic acid has many excellent characteristics and physiological functions. It has been used to treat abnormal immune activity, tumorigenesis and inflammation, and promote wound healing. It is also used in medicine, health food and Cosmetology and other fields have been extensively studied and widely used. Hyaluronic acid has different molecular weights, and its distribution ranges from several thousand to ...

Claims

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Application Information

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IPC IPC(8): C12N11/04C12N11/087C12N1/20C12P19/26C12R1/46
CPCC12N11/087C12N11/04C12N1/20C12P19/26C12N2513/00
Inventor 岳军崔镇华
Owner SUN YAT SEN UNIV SHENZHEN
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