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Protective agent for increasing stability of alpha-galactosidase prepared by aspergillus ZU-G1 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 improving the maximum efficiency, easy to use on a large scale, and broadening the application

Inactive Publication Date: 2009-07-29
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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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  • Protective agent for increasing stability of alpha-galactosidase prepared by aspergillus ZU-G1 foetidus fermentation and application thereof
  • Protective agent for increasing stability of alpha-galactosidase prepared by aspergillus ZU-G1 foetidus fermentation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Production of α-galactosidase crude enzyme liquid by liquid fermentation of Aspergillus futidatus ZU-G1

[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 for 15 minutes with a refrigerated centrifuge, and the rotating speed was 8000 rpm. The supernatant was collected and graded and salted out with ammonium sulfate (ammonium sulfate saturation 20%-70%), 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...

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]

Example Trehalose

(mg / 100ml

crude enzyme solution) Mannose

(mg / 100ml

crude enzyme solution) Raffinose

(mg / 100ml

crude enzyme solution) Enzyme activity retention *

(%) 2 37.8 18.22 75.6 67.95 3 37.8 27.33 100.8 72.71 4 37.8 36.44 126 64.19 ...

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% of 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]

[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 ice-bath it, and measure i...

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Abstract

The invention discloses a protective agent for improving and improving the stability of α-galactosidase produced by liquid fermentation of Aspergillus foetidus ZU-G1. The composition in parts by weight is: mannitol 18-36; raffinose 76-126; trehalose 38 ~95. When in use, 130-260 mg of the protective agent is added to every 100 ml of crude enzyme liquid of α-galactosidase produced by liquid fermentation of Aspergillus foetidus ZU-G1. Using the protective agent of the present invention, the enzyme activity retention rate of α-galactosidase produced by liquid fermentation of Aspergillus foetidus ZU-G1 is increased by 11.6% to 21.7% at 30°C to 60°C, and the acid resistance is reduced from pH4.0 to pH3.0 , 52% of the activity is retained in the environment of pH 3.0, which improves the maximum efficiency of α-galactosidase and broadens the application of the enzyme in acidic 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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Patent Type & Authority Patents(China)
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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