Preparation method for high purity fructo-oligosaccharide

A high-purity fructooligosaccharide technology, applied in directions such as being fixed on/in an organic carrier, fermentation, etc., can solve the problems of difficulty in industrialized production, expensive catalase, and no reuse of glucose oxidase, Achieve the effect of reducing production cost, low price and improving stability

Active Publication Date: 2009-02-18
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

The weakness of this double-enzyme method is that industrial production is difficult to rea

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Preparation of immobilized fructosyltransferase:

[0044] Aspergillus terricola strains were selected and inserted into a 200-liter fermenter containing 5% sucrose, 3% soybean meal powder, and 0.3% corn flour medium, and fermented at 33°C for 24 hours to obtain a seed culture solution. In a 5000-liter fermenter, add a culture medium solution, which contains 5% sucrose, 3% soybean meal, and 0.3% corn flour. Sterilize at 121°C for 40 minutes, add 150 liters of seed culture solution, and culture at 33°C and 220rpm for 24 hours to obtain a fermentation liquid containing mycelium, centrifuge, separate and collect mycelium, wash and break the wall 1. Take the enzyme solution by centrifugation. After measuring the enzyme activity by HPLC, add the enzyme solution and macroporous resin into a 2000-liter reaction tank at a ratio of 1 g of carrier per 40 u of the enzyme solution, stir evenly, cool to 8° C., and absorb for 12 hours. Add glutaraldehyde to make the final concent...

Embodiment 2

[0053] (1) Preparation of immobilized fructosyltransferase

[0054] Aspergillus terricola strains were selected and inserted into a 200-liter fermenter containing 5% sucrose, 3% soybean meal powder, and 0.3% corn flour medium, and fermented at 33°C for 24 hours to obtain a seed culture solution. In a 5000-liter fermenter, add a culture medium solution, which contains 5% sucrose, 3% soybean meal, and 0.3% corn flour. Sterilize at 121°C for 40 minutes, add 150 liters of seed culture solution, and culture at 33°C and 220rpm for 24 hours to obtain a fermentation broth containing mycelium, centrifuge, separate and collect mycelium, wash, and break the wall , centrifuge to get the enzyme solution, after measuring the enzyme activity by HPLC, add the enzyme solution and macroporous resin into a 2000 liter reaction tank at a ratio of 1 g of carrier per 40 u of the enzyme solution, stir evenly, cool to 8°C, and absorb for 12 hours. Add glutaraldehyde to make the final concentration 0....

Embodiment 3

[0064] (1) The preparation of immobilized fructosyltransferase was performed according to Example 1 of ZL 01128345.9.

[0065] (2) Preparation of immobilized glucose oxidase:

[0066] The immobilized carrier was prepared according to the method of Example 2. The crude enzyme solution of commercially available glucose oxidase was taken, and the enzyme activity was determined by titration. Add 200,000 to 500,000 U of enzyme solution to the carrier (wet), add 20Kg of porous microsphere carrier (wet) and 400,000 to 1,000,000 U of enzyme solution, and shake in a constant temperature shaker at 25 to 35°C and 200rpm After 2-4 hours, put it in the freezer at 4°C overnight, centrifuge, precipitate, rinse repeatedly with deionized water, and spin dry by centrifugation to obtain the immobilized glucose oxidase preparation.

[0067] (3) Preparation of immobilized hydrogen peroxide mimetic enzyme

[0068] In a 200-liter reaction tank, add 100L of 0.1mol / L sodium hydroxide solution, add 5...

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Abstract

The invention relates to a preparation method of high purified fructo-oligosaccharide, in particular to a method for preparing the high purified fructo-oligosaccharide by using immobilized enzyme. The preparation method of the invention prepares immobilized fructosyltransferase, immobilized glucose oxidase and immobilized mimic hydrogen peroxidase; then prepared enzymes are used to prepare the high purified fructo-oligosaccharide through an interrupted or continuous production method. In the preparation method, cheap metalporphyrin compounds are used as the mimic hydrogen peroxidase to replace expansive catalase; the fructosyltransferase, the glucose oxidase and the mimic hydrogen peroxidase are all immobilized and all can be recycled and reused; the stability and the operating factor of the enzymes are improved; the production cost for preparing the high purified fructo-oligosaccharide is greatly reduced. The preparation method can use one step method to directly produce the high purified fructo-oligosaccharide from cane sugar.

Description

technical field [0001] The invention relates to a production method of high-purity fructo-oligosaccharides, in particular to a method for producing high-purity fructo-oligosaccharides by using immobilized enzymes. Background technique [0002] Fructooligosaccharides (FOS), also known as fructooligosaccharides or fructooligosaccharides, are fructooligosaccharides produced by combining 1 to 3 fructosyl groups with fructosyl groups in sucrose through β (2-1) glycosidic bonds. A mixture of sugar, kestose and kestose pentaose, etc. Among them, kestose, kestetose and kestose pentaose are non-digestible sugars, which have the function of bifidobacteria proliferation and are effective components of functional oligosaccharides. So far, the fructo-oligosaccharide product on the market is divided into two kinds: a kind of is the fructo-oligosaccharide (hereinafter referred to as FOS) of about 50% oligosaccharide content 50 ), wherein the content of oligosaccharides (three sugars + te...

Claims

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Application Information

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IPC IPC(8): C12P19/00C12P19/18C12N11/02C12N11/06C12N11/04
CPCC12N11/06C12P19/18
Inventor 魏远安曾昭政姚评佳曾宪纲谢庆武谢拥葵梁锦添曾宪经陈子健陈振鹏
Owner 量子高科(广东)生物有限公司
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