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Compound immobilization method for fructosyl transferase

A technology of fructosyltransferase, enzyme solution, applied in microorganism-based methods, biochemical equipment and methods, immobilized on/in organic carriers, etc.

Inactive Publication Date: 2015-04-29
BAOLINGBAO BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The immobilized enzyme obtained by using chitosan as a carrier and glutaraldehyde as a cross-linking agent is in the form of debris, which is beneficial to the enzymatic reaction, but it is not easy to recycle, and there are problems in molding and shaping

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0031] A composite immobilization method for fructosyltransferase, comprising the steps of:

[0032] (1) Inoculate the enzyme-producing strain, Aspergillus niger, into the fermentation medium, and culture it for 6 days at a temperature of 28-32°C, an aeration rate of 500ml / min, and a stirring speed of 150r / min, to obtain a fermentation broth;

[0033] Above-mentioned fermentation medium component is as follows, is weight percent:

[0034] Sucrose 5%, soybean meal 10%, balance water, initial pH 6.5;

[0035] (2) The fermented broth obtained in step (1) is broken and filtered through the cells to obtain the crude enzyme liquid of fructosyltransferase; wherein, the natural cracking method is used for the cell rupture, and the specific steps include: using a filter cloth to filter the fermented Step (1) Filter and collect the bacterial cells after fermentation and culture, wash the bacterial cells with sterile water for 3 times, insert the sterile water with the same volume as th...

example 2

[0042] A composite immobilization method for fructosyltransferase, comprising the steps of:

[0043] (1) Inoculate the enzyme-producing strain, Aspergillus niger, into the fermentation medium, and culture it for 5 days at a temperature of 28-32°C, an aeration rate of 1000ml / min, and a stirring speed of 110r / min, to obtain a fermentation broth;

[0044] Above-mentioned fermentation medium component is as follows, is weight percent:

[0045] Sucrose 10%, soybean meal 7%, balance water, initial pH 7.0;

[0046] (2) The fermented broth obtained in step (1) is broken and filtered through the cells to obtain the crude enzyme liquid of fructosyltransferase; wherein, the natural cracking method is used for the cell rupture, and the specific steps include: using a filter cloth to filter the fermented Step (1) Filter and collect the bacterial cells after fermentation and culture, wash the bacterial cells with sterile water 4 times, insert the sterile water with the same volume as the f...

example 3

[0053] A composite immobilization method for fructosyltransferase, comprising the steps of:

[0054] (1) Inoculate the enzyme-producing strain, Aspergillus niger, into the fermentation medium, and culture it for 4 days at a temperature of 28-32°C, an aeration rate of 1500ml / min, and a stirring speed of 80r / min, to obtain a fermentation broth;

[0055] Above-mentioned fermentation medium component is as follows, is weight percent:

[0056] Sucrose 15%, soybean meal 5%, balance water, initial pH 7.5;

[0057] (2) The fermented broth obtained in step (1) is broken and filtered through the cells to obtain the crude enzyme liquid of fructosyltransferase; wherein, the natural cracking method is used for the cell rupture, and the specific steps include: using a filter cloth to filter the fermented Step (1) Filter and collect the bacterial cells after fermentation and culture, wash the bacterial cells with sterile water 5 times, insert the sterile water with the same volume as the fe...

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PUM

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Abstract

The invention discloses a compound immobilization method for fructosyl transferase, and belongs to the field of biotechnologies. The technology has the advantages that fructosyl transferase extraction technological equipment is simple; the fructosyl transferase is embedded by calcium alginate after chitosan covalence immobilization; the problems of forming and integer of chitosan covalence immobilized enzyme can be solved; the immobilization efficiency of the enzyme is high (enzyme activity recovery rate reaches above 70%); the enzyme can be used repeatedly for more than 85 times during batch reaction.

Description

technical field [0001] The invention relates to a composite immobilization method of fructosyltransferase, which belongs to the field of biotechnology. Background technique [0002] Fructose-oligosaccharides refer to the general term for kestose, kestose and kestose pentaose and their mixtures formed by the combination of fructosyl in sucrose and 1 to 3 D-fructose through β-1,2 glycosidic bonds. Also known as fructooligosaccharides, sucrose trisaccharide oligosaccharides, etc., the molecular formula is G-F-Fn (G is glucose, F is fructose, n=1~3). Fructose-oligosaccharide is a kind of prebiotics, which has physiological effects such as improving intestinal flora, inhibiting intestinal corruption, laxative, promoting mineral absorption, improving lipid metabolism, and enhancing immunity. At present, many enterprises at home and abroad use the enzymatic conversion method with sucrose as raw material to produce fructooligosaccharides. [0003] The principle of enzymatic conver...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/04C12R1/685C12R1/69
Inventor 刘宗利王乃强刘峰李克文王京博栾庆民张姝郝荣治张莉
Owner BAOLINGBAO BIOLOGY
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