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Method for preparing sialic acid by adopting immobilized enzyme

A technology for immobilizing enzymes and sialic acid, applied in the field of bioengineering, can solve the problems of easily polluted environment, complicated operation, large pollution, etc., and achieve the effects of less environmental pollution, mild reaction conditions and good effect.

Inactive Publication Date: 2017-02-15
袁春华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problems mainly solved by the present invention: when using traditional methods to prepare sialic acid at present, the reaction conditions of the chemical synthesis method are harsh, the yield is low, the cost of the enzymatic method is high, the operation is cumbersome and easy to pollute the environment, and the yield of other natural product extraction methods is low , which requires special equipment and is also highly polluted, provides a method for preparing sialic acid with immobilized enzymes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0019] First collect 150g of silkworm cocoons, and after spinning, mix it with a keratinase solution with a solid-to-liquid ratio of 1:5 and a concentration of 3g / L. After mixing, adjust the pH to 4.0 with a 25% oxalic acid solution, and heat the mixed solution to 80°C, heat preservation for 1 hour, suction filter to obtain filter residue, then mix the filter residue with 25% sodium hydroxide solution at a solid-to-liquid ratio of 1:5, heat to 75°C, heat preservation for 30 minutes for degumming, after degumming is completed, suction filter and collect the degumming The silk was rinsed with deionized water for 3 times, put into an oven, and dried at 90°C for 3 hours to obtain degummed silk; then the degummed silk was crushed and sieved to obtain a 100-mesh powder, which was mixed with a solid-liquid ratio of 1:60 Mix the sodium carbonate solution with a mass concentration of 2%, put it in a water bath at 70°C, stir and dissolve it for 30 minutes, inject the solution into a dial...

example 2

[0022] First collect 180g silkworm cocoons, mix them with a keratinase solution with a concentration of 3.5g / L at a solid-to-liquid ratio of 1:5 after spinning, adjust the pH to 4.5 with a 25% oxalic acid solution after mixing, and heat the mixture after adjustment to 85°C, heat preservation and degumming for 1.5h, and filter residue by suction filtration, and then mix the filter residue with 25% sodium hydroxide solution at a solid-to-liquid ratio of 1:5, heat to 80°C, heat preservation and degumming for 40 minutes, and suction filter after degumming is completed. Collect the degummed silk, wash it with deionized water for 4 times, put it in an oven, and dry it at 95°C for 4 hours to obtain the degummed silk; then crush the above degummed silk and sieve it to obtain a 110-mesh powder, and divide it according to the solid-to-liquid ratio of 1: 60 was mixed with 2% sodium carbonate solution with a mass concentration of 2%, put it into a water bath at 75°C, stirred and dissolved ...

example 3

[0025] First collect 200g silkworm cocoons, mix them with a keratinase solution with a solid-liquid ratio of 1:5 and a concentration of 4g / L after spinning, adjust the pH to 5.0 with a 25% oxalic acid solution after mixing, and heat the mixed solution to Degumming at 90°C for 2 hours, suction filtration to obtain filter residue, then mixing the residue with 25% sodium hydroxide solution at a solid-to-liquid ratio of 1:5, heating to 85°C, heat preservation for 50 minutes for degumming, after degumming is completed, suction filtration and collection of degumming The silk was rinsed with deionized water for 5 times, put into an oven, and dried at 100°C for 5 hours to obtain degummed silk; then the degummed silk was crushed and sieved to obtain a 120-mesh powder, which was mixed with a solid-liquid ratio of 1:60 Mix the sodium carbonate solution with a mass concentration of 2%, put it in a water bath at 80°C, stir and dissolve it for 40 minutes, inject the solution into a dialysis ...

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Abstract

The invention discloses a method for preparing sialic acid by adopting an immobilized enzyme, belonging to the technical field of bioengineering. The method comprises the following steps: dissolving silks after degumming in sodium carbonate solution, carrying out dialysis and drying to obtain fibroin, carrying out crosslinking and curing on the fibroin, isomerases and sialic acid aldolase, thus obtaining the immobilized enzyme, mixing the immobilized enzyme with whey liquid after degreasing, carrying out enzymic catalytic reaction, thus obtaining enzymatic hydrolysate, carrying out centrifugal separation to obtain a supernate, carrying out alcohol precipitation to obtain a sialic acid crude product, re-dissolving the sialic acid crude product in deionized water, removing insoluble impurities, carrying out filtering by adopting a membrane, and carrying out freeze drying to obtain the sialic acid product. The method has the advantages that the reaction conditions are mild, in the whole preparation process, the pollution to the environment is small, the extraction efficiency is high, the operation process is simple and novel, the prepared sialic acid is good in yield and product quality, the yield reaches up to 80% or above, the purity reaches up to 98% or above, and the obtained sialic acid can be widely applied to the technical fields of cosmetics, medicines and other biological medicines.

Description

technical field [0001] The invention discloses a method for preparing sialic acid by immobilized enzyme, which belongs to the technical field of bioengineering. Background technique [0002] Sialic acid is a family of neuraminic acid derivatives, the most common being N-acetylneuraminic acid, usually in the form of oligosaccharides, glycolipids or glycoproteins. The main food source of sialic acid is breast milk, but sialic acid is also found in milk, eggs, and cheese. In recent years, the application of sialic acid and its derivatives in food, health care products and medicine has increasingly broad development prospects, especially in infant formula milk powder. Studies have found that exogenous sialic acid can be supplemented by diet to Increase the content of sialic acid in the brain. This also indicates that adding sialic acid in infant milk powder, especially infant milk powder for premature infants, will effectively promote the development of their nervous system an...

Claims

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Application Information

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IPC IPC(8): C12N11/02C12P19/26C12P19/24
CPCC12N11/02C12N9/88C12N9/90C12P19/24C12P19/26
Inventor 袁春华张明王统军
Owner 袁春华
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