Method for preparing galactooligosaccharide by use of permeable cell beta-galactosidase

A technology of galactosidase and galacto-oligosaccharide is applied in the field of preparation of galacto-oligosaccharide, which can solve the problem that the product cannot be used directly, and achieve the effects of low production cost, reduced production cost and easy separation and recovery.

Inactive Publication Date: 2013-03-27
TIANJIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent CN 1225389A uses absolute ethanol and chloroform to obtain permeabilized Kluyveromyces lactis cell β-galactosidase through freez

Method used

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  • Method for preparing galactooligosaccharide by use of permeable cell beta-galactosidase
  • Method for preparing galactooligosaccharide by use of permeable cell beta-galactosidase
  • Method for preparing galactooligosaccharide by use of permeable cell beta-galactosidase

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Experimental program
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Embodiment 1

[0027] Preparation of Kluyveromyces lactis permeabilized cells: Fermentation experiments were carried out on Kluyveromyces lactis. The medium composition was: 3.0g yeast extract, 3.0g malt extract, 5.0g peptone, 20.0g lactose, 1.0L distilled water, pH 7.0, autoclave at 121°C for 20min. The fermentation conditions were: 28°C, shaker speed 100rpm, inoculum size 5% (v / v), cultured for 8 hours, and the enzyme activity of the obtained Kluyveromyces lactis fermentation broth was about 1.00U / mL. The fermentation broth was centrifuged at 5000 rpm and 4°C for 5 min, the precipitate was washed with water and then diluted to 50 g / L with PBS buffer to obtain suspension cells. Add absolute ethanol according to the volume ratio of absolute ethanol to suspension cell diluent at 2:5, and shake at 25°C for 1 min. After centrifuging at 5000rpm and 4°C for 5min, the pellet was washed twice with PBS buffer, the supernatant was discarded, and the pellet was taken to obtain the permeabilized cell ...

Embodiment 2

[0030] Preparation of Kluyveromyces lactis permeabilized cells: Fermentation experiments were carried out on Kluyveromyces lactis. The medium composition was: 3.0g yeast extract, 3.0g malt extract, 5.0g peptone, 20.0g lactose, 1.0L distilled water, pH 7.0, autoclave at 121°C for 20min. The fermentation conditions were: 30°C, shaker speed 160rpm, inoculum size 8% (v / v), cultured for 12h, and the enzyme activity of the obtained Kluyveromyces lactis fermentation broth was about 7.00U / mL. The fermentation broth was centrifuged at 5000 rpm and 4°C for 5 min, the precipitate was washed with water and then diluted to 50 g / L with PBS buffer to obtain suspension cells. Add absolute ethanol according to the volume ratio of absolute ethanol to suspension cell diluent at 4:5, and shake at 25°C for 10 min. After centrifuging at 5000rpm and 4°C for 5min, the pellet was washed twice with PBS buffer, the supernatant was discarded, and the pellet was taken to obtain the permeabilized cell β-g...

Embodiment 3

[0033] Preparation of Kluyveromyces lactis permeabilized cells: Fermentation experiments were carried out on Kluyveromyces lactis. The medium composition was: 3.0g yeast extract, 3.0g malt extract, 5.0g peptone, 20.0g lactose, 1.0L distilled water, pH 7.0, autoclave at 121°C for 20min. The fermentation conditions are: 30°C, shaker speed 250rpm, inoculum size 10% (v / v), culture for 20h. The enzyme activity of the obtained Kluyveromyces lactis fermentation broth was about 8.69U / mL. The fermentation broth was centrifuged at 5000 rpm and 4°C for 5 min, the precipitate was washed with water and then diluted to 50 g / L with PBS buffer to obtain suspension cells. Add absolute ethanol according to the volume ratio of absolute ethanol and suspension cell diluent at 1:1, and shake at 25°C for 15 minutes. After centrifuging at 5000rpm and 4°C for 5min, the pellet was washed twice with PBS buffer, the supernatant was discarded, and the pellet was taken to obtain the permeabilized cell β-...

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Abstract

The invention relates to a method for preparing galactooligosaccharide by use of permeable cell beta-galactosidase, which comprises the following steps of: adding permeable cell beta-galactosidase into a lactose solution, wherein the concentration of the lactose solution is 100-500g/L, and the addition amount of permeable cell beta-galactosidase is 200-1,500mg per gram of lactose; reacting for 2-15 hours at 30-45 DEG C; and performing centrifugal separation, wherein the obtained supernate is a galactooligosaccharide solution. By preparing galactooligosaccharide by use of permeable cell beta-galactosidase, the extraction and purification processes of endoenzyme are avoided, the technological operation is simplified, and the production cost is reduced. Kluyveromyces lactis is processed by absolute ethyl alcohol to prepare a permeable cell, toxic chemical reagents such as chloroform and the like are not introduced, the food safety problem is avoided, and the permeable cell can be widely applied to the food industry. Moreover, the permeable cell can be used repeatedly, thus the production cost of galactooligosaccharide can be obviously reduced.

Description

technical field [0001] The invention relates to a method for preparing galactooligosaccharides, in particular to a method for preparing galactooligosaccharides by using permeabilized cell β-galactosidase, and belongs to the fields of enzyme preparation preparation and functional oligosaccharide production. technical background [0002] With the change of the public diet structure, the demand for foods with the function of regulating physiological activities——functional foods is increasing, among which functional oligosaccharides are especially favored by people. my country launched the "National Public Nutrition Improvement OLIGO (Oligosaccharides) Project" in 2007, which greatly increased the market demand for functional oligosaccharides. Among them, galactooligosaccharides can be used to improve food color, flavor, texture and prolong shelf life because of their good water solubility, low viscosity, and stability to heat and acid. Fidobacterium can improve the distributio...

Claims

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Application Information

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IPC IPC(8): C12P19/14C12N9/38C12R1/645
Inventor 齐崴王梦凡何志敏张帅帅邢肖肖苏荣欣
Owner TIANJIN UNIV
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