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Method for preparing galacto-oligosaccharide

A technology of galactooligosaccharides and galactosidase, which is applied in the field of preparation of galactooligosaccharides, can solve the problem of less β-galactosidase, and achieve the effects of simplifying the separation and purification process, reducing pollution, and stabilizing physical and chemical properties

Active Publication Date: 2012-09-19
量子高科(广东)生物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is known that the β-galactosidase produced by Bacillus circulans is an extracellular enzyme and has the property of high temperature resistance, but there are few reports on the production of β-galactosidase by Bacillus circulans

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  • Method for preparing galacto-oligosaccharide

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preparation example Construction

[0032] The invention provides a kind of preparation method of galactooligosaccharide, comprises the following steps:

[0033] (1) Use glutaraldehyde to react with macroporous weak base styrene resin under stirring for 16-24 hours at 15-35°C to activate the resin;

[0034] (2) Take liquid β-galactosidase and the activated macroporous weak base styrene resin obtained in step (1) and stir and react at 15-35°C for 16-24 hours to obtain immobilized β-galactosidase;

[0035] (3) Add the immobilized β-galactosidase into the lactose solution, and stir and react at 48-52°C for 24-36h.

[0036] The present invention also provides a method for preparing galactooligosaccharides, the steps of which include the preparation of liquid β-galactosidase, the preparation of immobilized β-galactosidase and the production of oligosaccharides by immobilized β-galactosidase Three parts of galactose. Each section is described below.

[0037] 1. Preparation of liquid β-galactosidase

[0038]Using ...

Embodiment 1

[0049] The preparation of embodiment 1β-galactosidase

[0050] 1. Strain activation: inoculate Bacillus circulans preserved in glycerol tubes into LB solid medium, and incubate at 37°C for 18 hours.

[0051] 2. Seed liquid culture: After the Bacillus circulus is activated on the LB plate, pick a ring of bacteria and inoculate it into a Erlenmeyer flask filled with 25mL LB medium, shake and cultivate at 37°C and 180rpm for 4 hours, and then transfer to In a fermenter equipped with 2.5L LB medium, cultivate for 4 hours at 37°C, stirring speed 120-220rpm, ventilation rate 1.25L / min, control the dissolved oxygen ≥ 30% by adjusting the rotation speed, and obtain the seed culture solution.

[0052] 3. Fermentation culture:

[0053] 1) Pre-fermentation: Inoculate the seed culture solution into a mixture containing 1250g of lactose, 2000g of galactose, 4000g of soybean peptone, 1300g of corn extract, 650g of yeast extract, 650g of diammonium hydrogen phosphate, 325g of sodium carbona...

Embodiment 2

[0057] The immobilization of embodiment 2β-galactosidase

[0058] Follow the steps below to prepare immobilized β-galactosidase:

[0059] 1. Refining of crude enzyme

[0060] Take 100mL of the enzyme solution obtained by concentration in Example 1, slowly add 43.6g of ammonium sulfate to it, and after the ammonium sulfate is completely dissolved, let it stand at 4°C for 2 hours, then centrifuge at 10,000rpm for 10min at 4°C to collect the precipitate, and then use 100mL The precipitate was dissolved in pH6.0 phosphate buffer to obtain a refined solution of β-galactosidase with an enzyme activity of 100U / mL.

[0061] 2. Enzyme immobilization

[0062] (1) Weigh 100g of macroporous weak base styrene resin A103s, wash it with deionized water for 2~3 times, add 2000mL of deionized water, then add 40mL of 50% glutaraldehyde solution, stir at 30°C for 16h, filter, then Clean the resin with deionized water, filter and dry the water to obtain the activated resin.

[0063] (2) Add 1...

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Abstract

The invention discloses a method for preparing galacto-oligosaccharide. The method comprises the following steps: (1) reacting glutaraldehyde with a macroporous weak basic styrene resin for 16 to 24 hours with stirring at the temperature of between 15 and 35DEG C to activate the resin; (2) reacting liquid beta-galactosidase with the activated macroporous weak basic styrene resin obtained in the step (1) for 16 to 24 hours with stirring at the temperature of between 15 and 35DEG C to obtain cured beta-galactosidase; and (3) adding the cured beta-galactosidase into a lactose solution, and reacting for 24 to 36 hours with stirring at the temperature of between 48 and 52DEG C. By the method for producing the galacto-oligosaccharide, the filter pressure can be kept stable in a syrup filtering process, the yield of the galacto-oligosaccharide is improved, the energy consumption is reduced and the industrial production requirement can be met.

Description

technical field [0001] The invention relates to a method for preparing galactooligosaccharides, in particular to a method for producing galactooligosaccharides by using immobilized enzymes. Background technique [0002] Galacto-oligosaccharide (GOS) is a kind of functional oligosaccharides naturally present in animal breast milk. It has excellent water retention capacity and strong acid and heat stability. It cannot be digested and absorbed by human small intestine, but can Fermented by colonic flora, it can promote the proliferation of bifidobacteria, inhibit the growth of harmful pathogenic bacteria and spoilage bacteria, promote the absorption of calcium, magnesium, and potassium, facilitate the production of B vitamins, niacin, and folic acid, and effectively stimulate intestinal peristalsis. Reduce and prevent constipation, reduce the content of toxic substances in the colon, regulate intestinal microecology, and promote intestinal health. [0003] Galactooligosacchari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12N11/08C12R1/09
Inventor 曾宪经曾宪纲陈子健陈振鹏肖桂秋
Owner 量子高科(广东)生物有限公司
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