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A method for producing d-tagatose

A technology for tagatose and lactose, which is applied in the field of functional rare sugar preparation, can solve the problems of high cost of separation and purification, difficulty in separation and purification of tagatose, etc., and achieves the effects of low cost, easy operation and cost reduction

Active Publication Date: 2020-11-20
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the closeness of the properties of glucose and tagatose, it is extremely difficult to separate and purify tagatose products in the later stage, and the cost of separation and purification is high
[0005] After retrieval, the use of low-cost lactose as a raw material to produce D-tagatose, and the simple method of enzymatically converting lactose hydrolysis by-product glucose into negatively charged gluconic acid and using ion exchange resins for adsorption and removal have not been reported.

Method used

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  • A method for producing d-tagatose
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Embodiment 1

[0053] The preparation method of embodiment 1 immobilized β-galactosidase

[0054] (1) Add concentrated hydrochloric acid according to the ratio of 20mL concentrated hydrochloric acid to every 5g of silica gel, use a rotary evaporator to reflux at 86°C for 2.5h, 8000rpm, and centrifuge for 5min to remove the supernatant;

[0055] (2) Using 25mL of distilled water, stir at 67°C and 100rpm for 2h, and wash the silica gel obtained in step (1);

[0056] (3) Add 2 mL of 2% NaOH to the silica gel obtained in step (2), stir at 40° C. at 100 rpm for 1.5 h, then centrifuge at 8000 rpm for 5 min to remove the supernatant;

[0057] (4) Add 25mL activation solution (7mL 0.8mol / LNaOH, 10mL dimethyl sulfoxide, 8mL epichlorohydrin) to the silica gel obtained in step (3), activate at 40°C for 2.5h at 170rpm, and centrifuge at 8000rpm for 5min to remove the upper Serum;

[0058] (5) Add 80 mL of amination solution (60 mL of tetrahydrofuran, 20 mL of 3-aminopropyltriethoxysilane) to the silic...

Embodiment 2

[0066] Embodiment 2 Preparation method of immobilized L-arabinose isomerase

[0067] (1) Pretreatment of anion exchange resin: put the resin into a chromatographic column with an inner diameter of 3 cm, the height of the column bed is 30 cm, and wash it repeatedly with pure water until there are no visible mechanical impurities in the sample and the water is clear;

[0068] (2) Use 100mL of 1N hydrochloric acid solution, 200mL of pure water, 100mL of 1N sodium hydroxide solution and 200mL of pure water to pass through the resin layer from top to bottom. When switching reagents each time, make the liquid level 1cm higher than the resin layer to ensure that there are no air bubbles in the resin layer; perform this operation twice;

[0069] (3) Pass through the resin layer with 400mL 1N sodium hydroxide solution through the resin treated in step (2), the flow rate is 10mL / min, then wash with pure water until the effluent is colorless when tested with phenolphthalein indicator sol...

Embodiment 3

[0075] The preparation method of embodiment 3 D-tagatose conversion liquid

[0076] (1) Prepare a lactose solution with a final concentration of 100 g / L with a 10 mM phosphate buffer solution having a pH of 6.5;

[0077] (2) Mix immobilized β-galactosidase, immobilized glucose oxidase and immobilized L-arabinose isomerase in a weight ratio of 1:50:8 to prepare a composite immobilized enzyme and use it as a solid catalyst Add the solid catalyst to the lactose solution at a rate of 500 g of catalyst per 1 L of the lactose solution prepared in step (1), react at 55°C for 3 hours at 100 rpm, then centrifuge at 10,000 rpm for 5 minutes to remove the solid catalyst;

[0078] (3) Add macroporous styrene-based weakly basic anion exchange resin D309 to the lactose solution at a rate of 500 g of anion exchange resin per 1 L of the lactose solution prepared in step (2), stir for 3 hours at 55°C, 100 rpm, and use The anion-exchange resin is removed by filtration through a filter membran...

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Abstract

The invention discloses a method for producing D-tagatose at low cost. The method comprises the following steps: taking lactose as a raw material; meanwhile, adding a compound enzymic preparation containing immobilized beta-galactosidase, L-arabinose isomerase and glucose oxidase, hydrolyzing the lactose into galactose and glucose by using the beta-galactosidase; catalyzing the galactose to generate the D-tagatose by using the L-arabinose isomerase; oxidizing a by-product glucose into gluconic acid with negative charge by using the glucose oxidase; removing the gluconic acid by using ion exchange resin, and removing the by-product, thereby obtaining transformed liquid containing the D-tagatose. The method disclosed by the invention has the advantages of simple process, convenience in operation, high efficiency, capabilities of reducing production cost of the D-tagatose and broad industrial production and application prospects.

Description

technical field [0001] The invention relates to a method for producing D-tagatose, in particular to a method for producing D-tagatose by using lactose as a raw material, which belongs to the technical field of functional rare sugar preparation. Background technique [0002] Rare sugars are an important class of carbohydrates, which play very important functions in the fields of diet, health care, and medicine. According to the definition of the International Sugar Association, rare sugars are "a class of monosaccharides and their derivatives that exist in nature but are very low in content", generally have the characteristics of low calorie, low absorption, etc., and have a variety of physiological functions. [0003] D-Tagatose (abbreviated as D-Tag) is a six-carbon ketose sugar with a sweetness of 92% of that of sucrose, and the heat generated is only 1 / 3 of that of sucrose. In view of the good properties of D-tagatose, the US Food and Drug Administration confirmed the sa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/02C12N11/14C07H3/02C07H1/06
CPCC07H1/06C07H3/02C12N9/2471C12N9/90C12N11/14C12P19/02C12Y302/01023C12Y503/01004
Inventor 林建群李灿郭青青林建强
Owner SHANDONG UNIV
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