A kind of preparation method of α-2,3 deaminosialyllactulose

A technology of lactulose and sialic acid, applied in the field of α-2, can solve problems such as the difficulty of deaminosialic acid glycosylation

Active Publication Date: 2021-03-19
河南省熙康食品有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the invention is to overcome the difficult problem of deaminosialic acid glycosylation of lactulose by chemical methods

Method used

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  • A kind of preparation method of α-2,3 deaminosialyllactulose
  • A kind of preparation method of α-2,3 deaminosialyllactulose
  • A kind of preparation method of α-2,3 deaminosialyllactulose

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Embodiment 1

[0014] A method for preparing α-2,3 deaminosialyllactulose, characterized in that: 40 parts by weight of mannose, 100 parts by weight of sodium pyruvate, 50 parts by weight of lactulose, and 150 parts by weight of cytidine triphosphate sodium (CTP) , dissolved in 4000 parts by weight of ultrapure water, adjusted to pH 8.5 with 4 mol / liter of NaOH, adding 400 parts by weight of Tris-HCl of 1 mol / liter of pH 8.5; adding 40 parts by weight of 2 mol / liter of MgCl 2 ; Add 5 parts by weight of sialic acid aldolase, 6 parts by weight of CMP-sialyltransferase; 6 parts by weight of α-2,3 sialyltransferase. Then the mixture was placed in a 37°C air bath shaker and reacted for 24 hours. The degree of reaction was detected by silica gel thin layer chromatography (TLC), and the developing solvent of TLC was n-propanol:methanol:water=4:2:1. After the lactulose reaction is complete, add the same volume of 95% ethanol to the reaction solution, mix well, put it in a refrigerator at 4°C for 30...

Embodiment 2

[0016] A method for preparing α-2,3 deaminosialyllactulose, characterized in that: 80 parts by weight, 200 parts by weight of sodium pyruvate, 100 parts by weight of lactulose, 260 parts by weight of cytidine triphosphate sodium (CTP), dissolved in In 8000 parts by weight of ultrapure water, adjust the pH to 8.5 with 4 mol / liter of NaOH, add 1000 parts by weight of Tris-HCl of 1 mol / liter of pH 8.5; add 100 parts by weight of MgCl of 2 mol / liter 2 ; Add 8 parts by weight of sialic acid aldolase, 10 parts by weight of CMP-sialyltransferase; 10 parts by weight of α-2,3 sialyltransferase. Then the mixture was placed in a 37°C air bath shaker and reacted for 24 hours. The degree of reaction was detected by silica gel thin layer chromatography (TLC), and the developing solvent of TLC was n-propanol:methanol:water=4:2:1. After the lactulose reaction is complete, add the same volume of 95% ethanol to the reaction solution, mix well, put it in a refrigerator at 4°C for 30 minutes, th...

Embodiment 3

[0018] A method for preparing α-2,3 deaminosialyllactulose, characterized in that: 50 parts by weight of mannose, 120 parts by weight of sodium pyruvate, 65 parts by weight of lactulose, and 160 parts by weight of cytidine triphosphate sodium (CTP) , dissolved in 6000 parts by weight of ultrapure water, adjusted to pH 8.5 with 4 mol / liter of NaOH, adding 600 parts by weight of Tris-HCl of 1 mol / liter of pH 8.5; adding 60 parts by weight of 2 mol / liter of MgCl 2 ; Add 6 parts by weight of sialic acid aldolase, 7 parts by weight of CMP-sialyltransferase; 7 parts by weight of α-2,3 sialyltransferase. Then the mixture was placed in a 37°C air bath shaker and reacted for 24 hours. The degree of reaction was detected by silica gel thin layer chromatography (TLC), and the developing solvent of TLC was n-propanol:methanol:water=4:2:1. After the lactulose reaction is complete, add the same volume of 95% ethanol to the reaction solution, mix well, put it in a refrigerator at 4°C for 30...

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Abstract

The invention discloses a method for preparing alpha-2,3 deaminosialyl lactulose. The method is characterized in that 40-80 parts by weight of mannose, 100-200 parts by weight of sodium pyruvate, 50-100 parts by weight of lactulose and 125-260 parts by weight of sodium cytidine triphosphate (CTP) are dissolved in 4000-8000 parts by weight of ultrapure water, the pH is adjusted to 8.5 with 4 mol / liter of NaOH, and 400-1000 parts by weight of 1 mol / liter of Tris-HCl with pH of 8.5 are added; 40-100 parts by weight of 2 mol / L of MgCl2 are added; 5-8 parts by weight of sialic acid aldolase areadded, and 6-10 parts by weight of CMP-sialyltransferase are added; 6-10 parts by weight of alpha-2,3 sialyltransferase are added. Reaction is conducted for 24 hours at 37 DEG C in a vapour-bathing constant temperature vibrator. After lactulose is completely reacted, 95% of ethanol of the same volume is added to a reaction solution, the reaction solution is placed in a refrigerator at 4 DEG C for30 min, and then is centrifuged at 7000 rpm for 30 min, and supernatant is concentrated through rotary evaporation. Purification is conducted with biogel columns, concentration is conducted through the rotary evaporation, and pure products are obtained through freezing and drying.

Description

technical field [0001] The invention belongs to the technical field of preparation of functional oligosaccharides, and in particular relates to a preparation method of α-2,3 deaminosialyllactulose. Background technique [0002] Sialic acid (Sialic acid) is a general term for acyl derivatives of nine-carbon monosaccharide compounds with carboxyl groups. It is widely found in many organisms such as bacteria, fish, and mammals. The end of the sugar chain participates in and regulates many important life activities, such as cell recognition, biofilm flow, and endocytosis, and is an important molecule in higher animals and some microorganisms. There are nearly 50 known members of sialic acid, among which, N-acetylneuraminic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc) and deaminoneuraminic acid (KDN) are the three components of sialic acid. A core monomer, and the rest of the sialic acid is derived from these three monomers. [0003] Desaminosialic acid (KDN) is one of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/18C12P19/00
CPCC12P19/00C12P19/18
Inventor 曾洁张瑞瑶贾甜胡雅婕高海燕孙俊良李光磊
Owner 河南省熙康食品有限公司
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