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Method for preparing high-purity crystallized D-Tagatose

A tagatose, cooling crystallization technology, applied in the preparation of sugar derivatives, chemical instruments and methods, monosaccharides, etc., can solve the problems of undeveloped research on the separation and purification of tagatose

Active Publication Date: 2021-04-02
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the separation and purification of tagatose in the above-mentioned multi-enzyme catalysis and whole-cell catalysis system has not yet been studied.

Method used

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  • Method for preparing high-purity crystallized D-Tagatose
  • Method for preparing high-purity crystallized D-Tagatose
  • Method for preparing high-purity crystallized D-Tagatose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Preparation of starting material for S1 purification:

[0035] The multi-enzyme catalytic reaction system contains 30 mM phosphate buffer (pH 7.0), 5 mM divalent magnesium ions, the amount of α-glucan phosphorylase is 10 U / mL, the amount of glucose phosphomutase 10 U / mL, the dosage of glucose phosphate isomerase is 10 U / mL, the dosage of 6-phosphate tagatose epimerase is 15 U / mL, the dosage of 6-phosphate tagatose phosphatase is 15 U / mL, the dosage of starch debranching enzyme is 1 U / mL, soluble starch is 70 g / L, and the catalytic reaction is carried out at 70°C. After 36 hours of reaction, 10% sulfuric acid is added to terminate the reaction, and the detection is carried out by HPLC. Among them, the HPLC detection conditions are as follows: the chromatographic column is Bio-Rad HPX-87H; the flow rate is 0.6 mL / min; the column temperature is 60°C; the detector is a differential refractive index detector; the injection volume is 20 μL. The composition of the obtained re...

Embodiment 2

[0046] Preparation of starting material for S1 purification:

[0047] The whole-cell catalytic reaction system contains 30 mM phosphate buffer (pH 7.0), 5 mM divalent magnesium ions, the dosage of cells expressing α-glucan phosphorylase is 1.5 g DCW / mL, and the amount of cells expressing glucose phosphorylase is 1.5 g DCW / mL. 0.8 g DCW / mL for cells expressing bitase, 0.8 g DCW / mL for cells expressing glucose phosphoisomerase, and 6 g DCW for cells expressing tagatose-6-phosphate epimerase / mL, the amount of cells expressing 6-phosphate tagatose phosphatase is 6 gDCW / mL, the amount of cells expressing starch debranching enzyme is 0.8 g DCW / mL, maltodextrin 135 g / L, at 70 ℃ Catalyzed reaction, after 48 hours of reaction, the composition of the obtained reaction solution is as follows: tagatose 95.6 g / L, glucose 5.02 g / L, maltobiose 25.4 g / L, oligosaccharides (DP≥3) ~13 g / L , inorganic salts, pigments, cells and proteins. Carry out HPLC measurement by the same condition in embo...

Embodiment 3

[0059] Preparation of starting material for S1 purification:

[0060] The immobilized whole-cell catalytic reaction system contains 30 mM phosphate buffer (pH 7.0), 5 mM divalent magnesium ions, and simultaneously immobilizes cells expressing α-glucan phosphorylase and expressing glucose phosphomutase Cells expressing glucose phosphate isomerase, cells expressing 6-phosphate tagatose epimerase, and cells expressing 6-phosphate tagatose phosphatase were used as catalysts, and the amount of immobilized cells was 20 g / L, maltodextrin 210 g / L, catalytic reaction at 70°C, after 48 hours of reaction, the composition of the obtained reaction liquid is as follows: tagatose 120 g / L, glucose 7.93 g / L, maltobiose 18.0 g / L, oligosaccharides (DP≥3) ~65 g / L, inorganic salts, pigments. The HPLC diagram of each component is shown in Figure 6 shown.

[0061] Since bacteria and proteins do not exist in the immobilized whole-cell catalytic reaction system, no bacteria and / or proteins are sep...

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Abstract

The invention relates to a method for separating and purifying D-Tagatose from a multi-enzyme catalysis and / or whole-cell catalysis (including immobilized enzymes and / or immobilized cells) system. Themethod comprises the following steps: by taking a D-Tagatose reaction liquid as an initial raw material, decoloring, desalting, performing chromatographic separation and crystallizing, and finally obtaining a high-quality D-Tagatose crystal product with the total separation and purification yield of not less than 80%, wherein the purity of the D-Tagatose crystal product is greater than or equal to 99%. According to the method disclosed by the invention, full-value utilization of a substrate is realized by recycling the substrate separated on the basis of a simulated moving bed, the utilization rate of ion exchange resin is improved by desalting through a continuous ion exchange system, and the production cost is reduced by virtue of a separation and purification process of ethanol coolingcrystallization based on ethanol recycling. The separation and purification yield of the D-Tagatose in the separation and purification process is high, the obtained D-Tagatose product is high in quality, and industrial production of the D-Tagatose is promoted.

Description

technical field [0001] The invention belongs to the field of organic analysis and purification, specifically relates to a method for preparing high-purity tagatose, more specifically relates to separating and purifying tagatose from a multi-enzyme catalysis and / or whole cell catalysis (including immobilized enzyme and / or immobilized cells) system sugar method. Background technique [0002] D-Tagatose is a six-carbon ketose sugar with the molecular formula C 6 h 12 o 6 , is the epimer of fructose and the aldehyde and ketone isomer of galactose. It is a rare monosaccharide that occurs naturally, mainly in dairy products such as yogurt and milk powder. Its sweetness properties are similar to sucrose, and the calories produced are only one-third of sucrose, so it is called a low-calorie sweetener. Tagatose was officially approved by the US Food and Drug Administration (US FDA) as a generally recognized as safe food (GRAS) in 2001, and was approved by the European Union in 2...

Claims

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Application Information

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IPC IPC(8): C07H3/02C07H1/06C12P19/02
CPCC07H1/06C07H3/02C12P19/02
Inventor 马延和石婷李元
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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