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Method for preparing high-purity galactooligosaccharide by beta-galactosidase

A technology of galacto-oligosaccharide and galactosidase, which is applied in microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of difficulty in industrialized production, expensive enzyme preparations, and low recycling rate. , to achieve mild and pollution-free fermentation conditions, reduce the consumption of preparation links, and achieve mild fermentation conditions.

Pending Publication Date: 2020-06-26
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are often limited by high cost, complicated operation requirements, and the product purity is usually less than 70%
Among them, the enzymatic treatment has good purification ability, but the enzyme preparation is expensive, the recycling rate is low, and it is not easy to operate, so it is difficult to realize further industrial production

Method used

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  • Method for preparing high-purity galactooligosaccharide by beta-galactosidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Free β-galactosidase catalyzes lactose to prepare crude galactooligosaccharides by transglycosidation reaction:

[0027] Prepare 400g / L lactose solution using 50mM phosphate buffer solution with pH = 8, take 20mL lactose solution and put it in a 50mL jar, dilute it 1000 times and measure the enzyme of free β-galactosidase enzyme solution by enzyme activity detection method The activity is 36U / mL, add free β-galactosidase enzyme solution according to the ratio of 30U / g lactose, react at 37°C, 160rpm constant temperature shaking water bath for 2h, after the reaction is completed, use boiling water to inactivate for 10min, and obtain the crude product of GOS . The crude GOS product was diluted with pure water to an appropriate multiple, filtered with a 0.45 μm filter membrane, and the content of each component in the product was detected by high-performance liquid chromatography according to HPLC detection conditions, and the yield of GOS was measured to be 22.78%.

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Embodiment 2

[0036] (1) Free β-galactosidase catalyzes lactose to prepare crude galactooligosaccharides by transglycosidation reaction:

[0037]Prepare 400g / L lactose solution using 50mM phosphate buffer solution with pH = 8, take 20mL lactose solution and put it in a 50mL jar, dilute it 1000 times and measure the enzyme of free β-galactosidase enzyme solution by enzyme activity detection method The activity is 36U / mL, add free β-galactosidase enzyme solution according to the ratio of 25U / g lactose, react at 37°C, 160rpm constant temperature shaking water bath for 2h, after the reaction is completed, use boiling water to inactivate for 10min, and obtain the crude product of GOS . The crude GOS product was diluted with pure water to an appropriate multiple, filtered through a filter membrane of 0.45 μm, and the content of each component in the product was detected by high-performance liquid chromatography according to HPLC detection conditions, and the yield of GOS was measured to be 21.04%...

Embodiment 3

[0044] (1) Free β-galactosidase catalyzes lactose to prepare crude galactooligosaccharides by transglycosidation reaction:

[0045] Prepare 500g / L lactose solution using 50mM phosphate buffer solution with pH=9, take 20mL lactose solution and place it in a 50mL jar, dilute it 1000 times and measure the enzyme activity of free β-galactosidase enzyme solution by enzyme activity detection method The activity is 36U / mL, add free β-galactosidase enzyme solution according to the ratio of 1U / g lactose, react in a constant temperature shaking water bath at 25°C and 160rpm for 2h, and inactivate with boiling water for 10min after the reaction is completed to obtain the crude product of GOS . The crude GOS product was diluted with pure water to an appropriate multiple, filtered through a filter membrane of 0.45 μm, and the content of each component in the product was detected by high-performance liquid chromatography according to HPLC detection conditions, and the yield of GOS was measu...

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Abstract

The invention relates to a method for preparing high-purity galactooligosaccharide by beta-galactosidase. The method comprises the following steps: (1) preparing galactooligosaccharide (GOS) by usingfree beta-galactosidase to catalyze lactose through a transglycosylation reaction; (2) performing whole-cell fermenting preparation with Kluyveromyces lactis; and (3) obtaining the high-purity galactooligosaccharide by whole-cell purification with the Kluyveromyces lactis. After a GOS synthesis is carried out for 2 hours, the GOS yield is 22.78%, and after whole-cell treatment with Kluyveromyces lactis is carried out for 15 hours, the purity of GOS is over 90%. The process provides a simple, efficient and inexpensive method for production of high-purity oligosaccharides. The GOS produced by the method is high in purity, low in production process cost and high in efficiency, is green and environmentally friendly, and is expected to be widely applied.

Description

technical field [0001] The invention relates to a method for preparing high-purity galactooligosaccharides by β-galactosidase, which belongs to the field of food biotechnology. Background technique [0002] With the rapid development of China's economy and society, the aging process is gradually accelerating, and the society's voice for the development of health food and functional food is also getting higher and higher. Galacto-oligosaccharides (GOS for short), as a new type of functional food, has broad market prospects in my country. Therefore, the development of a low-cost, high-efficiency, environmentally friendly, high-purity GOS green production process suitable for large-scale production is of great significance to the industrial production of GOS in my country. [0003] GOS has the characteristics of low calorific value and strong moisture retention, and has high thermal stability under acidic and neutral pH conditions. As a low-energy carbohydrate, it can be used...

Claims

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Application Information

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IPC IPC(8): C12P19/14C12P19/04C12N1/16C12R1/645
CPCC12P19/14C12P19/04C12N1/16
Inventor 齐崴尤生萍吴昊苏荣欣
Owner TIANJIN UNIV
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