Protective agent for increasing stability of alpha-galactosidase prepared by aspergillus foetidus fermentation and application thereof

A technology of galactosidase, ZU-G1, applied in the direction of enzyme stabilization, enzymes, enzymes, etc., can solve the problems of lack of theoretical basis for the effective application of liquid α-galactosidase preparations, few research reports on enzyme stability, etc. , to achieve the effect of being easy to use on a large scale, broadening applications and having a wide range of sources.

Inactive Publication Date: 2007-08-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Researchers at home and abroad have carried out preliminary research and exploration on technical issues such as the production and application of α-galactosidase, but there are few reports on the stability of the enzyme, and there is a lack of effective application of liquid α-galactosidase preparations. Theoretical basis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Aspergillus futidatus ZU-G1 liquid fermentation produces α-galactosidase crude enzyme liquid

[0026] According to the method disclosed in the Chinese invention patent application 200610050135.4, 30 ml of fermentation medium is filled in a 250 ml Erlenmeyer flask, and 1 ml of Aspergillus foetidus ZU-G1 spores (concentration of 1 × 10 6 individual / ml), the α-galactosidase fermentation broth obtained after culturing for 96 hours at 28° C. on a shaker with a rotational speed of 200 rpm. The fermented liquid was centrifuged with a refrigerated centrifuge for 15min, and the rotating speed was 8000rpm. The supernatant was collected and graded and salted out with ammonium sulfate (ammonium sulfate saturation 20%-70%), and then the precipitate was collected and washed with a small amount of 0.2mol pH5.0 / L disodium hydrogen phosphate-citric acid buffer solution, put it into a dialysis bag, and dialyze with distilled water for several hours (4°C), until no sulfate i...

Embodiment 2～10

[0028] Add the protective agent of the present invention in the crude enzyme liquid (enzyme activity about 100U / ml) that obtains in embodiment 1; The use ratio of mannitol, trehalose and raffinose in every 100ml crude enzyme liquid is shown in Table 1, crude enzyme liquid After adding the protective agent of the present invention, put it in a water bath at 60° C. for 1 hour, then immediately in an ice bath, and measure its enzyme activity after diluting 100 times. Among them, trehalose is produced by Shanghai Yuanju Biotechnology Co., Ltd.; mannitol is produced by Guangdong Taishan Chemical Factory; and raffinose is produced by American Amresco.

[0029] Table 1

[0030]

[0031] 5

[0032] The analysis of variance shows that the optimal formulation of the protective agent is: the weight-volume ratio of mannitol to crude enzyme solution is 27.33mg / 100ml, the weight-volume ratio of trehalose to crude enzyme solution is 66.15mg / 100ml, and the ratio of raf...

Embodiment 11

[0033] Example 11 Experiment of using the protective agent of the present invention to improve the thermal stability of α-galactosidase under different temperature conditions

[0034] The α-galactosidase produced by liquid fermentation of Aspergillus foetidus ZU-G1 added with a protective agent and the control group without a protective agent were calculated as 100% based on the enzyme activity retention rate without heat treatment and without a protective agent placed at 4°C. The enzyme activity retention rates obtained after heat treatment at different temperatures for different times were compared. In this experiment, 27mg of mannitol, 66mg of trehalose and 101mg of raffinose were added to every 100ml of crude enzyme solution. The specific results are shown in Table 2.

[0035] Table 2

[0036] temperature

[0037] After adding the protective agent of the present invention to the crude enzyme solution, put it in a water bath at 60°C for 1 hour, then immediately ...

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PUM

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Abstract

The invention discloses an alpha-galactosidase stability protective agent to increase and improve odor aspergillus ZU-G1 liquid fermentation production, which is characterized by the following: making mass percent as 18-36 wt manna sugar, 76-126 wt raffinose and 38-95 mycose; adding 130-260mg protective agent into per 100ml alpha-galactosidase rough enzyme liquid with odor aspergillus ZU-G1 liquid; increasing 30-60 deg.c enzyme active reserving ratio at 11.6 %-21.7% with this protective agent; reducing acid-resistivity from pH4.0 to pH3.0; retaining 52% activator under the environment of pH3.0; increasing biggest worked efficiency of alpha-galactosidase; widening the appliance of this enzyme in acid environment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protective agent for improving and improving the stability of α-galactosidase produced by liquid fermentation of Aspergillus foetidus ZU-G1 and its application. Background technique [0002] Soy products and their functional ingredients have good nutritional value and health functions, and are of high quality and low price, so they are indispensable food on the table of all grades. However, the oligosaccharides contained in beans are not easy to be digested and absorbed by the human body. It is fermented by bacteria in the intestinal tract, and it will cause belching, abdominal distension, abdominal pain and other human discomfort symptoms after eating. Research in recent years has found that α-galactosidase can solve this problem very well. α-Galactosidase can hydrolyze stachyose, raffinose, and melibiose in oligosaccharides to produce monosaccharides. It is beneficial to the abs...

Claims

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Application Information

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IPC IPC(8): C12N9/96C12N9/38
Inventor 刘彩琴何国庆傅伟陈启和阮晖
Owner ZHEJIANG UNIV
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