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Fermentation medium and method for improving activity of beta-glucuronidase produced by fungi

A technology of aldolidase enzyme activity and enzyme production medium, which is applied in the biological field and can solve the problems of low enzyme production activity and long enzyme production time

Inactive Publication Date: 2016-10-26
BEIJING INSTITUTE OF TECHNOLOGYGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, it has been found that there are many β-glucuronidases from fungi that can convert GL. For example, a strain of Penicillium purpurogenum Li-3 (Penicillium purpurogenum Li-3) Li-3 with a preservation number of CGMCC No. 5446 is recorded in the invention patent application 201410088328.3 Bacterial strain, a preservation number found by the inventor is the Talaromyces pinophilum (Talaromyces pinophilum) Li-93 bacterial strain of CGMCC No.11765, and the literature "Wang Xiaoyan Shihezi University Master's Thesis, 2007" reports Aspergillus terreus (Aspergillus terreus) bacterial strain Li- 20 and the Aspergillus ustus strain (Aspergillus ustus) Li-62, both of which can produce β-glucuronidase, which can convert GL to GAMG and / or GA, but these strains were fermented with existing media to produce β-glucuronidase. When glucuronidase is produced, there are disadvantages such as low enzyme activity and long enzyme production time, which limit the utilization of the enzyme

Method used

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  • Fermentation medium and method for improving activity of beta-glucuronidase produced by fungi

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Embodiment 1

[0039] Embodiment 1, adopting No. 1 fermentation enzyme production medium of the present invention to carry out fermentation production enzyme

[0040] Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20, and Aspergillus pyrogenus Li-62 were respectively fermented to produce enzymes. The specific operation process was as follows:

[0041] Inoculate the spores of Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20 and Aspergillus pylorus Li-62 in potato dextrose liquid medium respectively, and cultivate them on a shaker at 30°C and 200rpm 48h. Then insert 5% of the inoculum into the medium of the secondary seed liquid, culture on a shaker at 30° C. and 200 rpm for 24 hours to obtain the secondary seed liquid of each fungus mentioned above.

[0042] Secondary seed liquid culture medium: Weigh 200g of peeled potatoes, add 800mL of water, boil for about 30 minutes, filter through gauze, add 1g of glucose, dissolve and add water to 2000m...

Embodiment 2

[0045] Embodiment 2, adopt No. 2 fermentation enzyme production medium of the present invention to carry out fermentation production enzyme

[0046] Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20, and Aspergillus pyrogenus Li-62 were respectively fermented to produce enzymes. The specific operation process was as follows:

[0047] Inoculate the spores of Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20 and Aspergillus pylorus Li-62 in potato dextrose liquid culture medium respectively, at 30°C, 200rpm Incubate on a shaker for 48 hours. Then insert 5% of the inoculum into the medium of the secondary seed liquid, culture on a shaker at 30° C. and 200 rpm for 24 hours to obtain the secondary seed liquid of each fungus mentioned above.

[0048] Secondary seed liquid medium: Weigh 200g of peeled potatoes, add 800mL of water, boil for about 30 minutes, filter through gauze, add 5g of glucose, dissolve and add water to 2000mL, natu...

Embodiment 3

[0051] Embodiment 3, adopt No. 3 fermentation enzyme production medium of the present invention to carry out fermentation production enzyme

[0052] Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20, and Aspergillus pyrogenus Li-62 were respectively fermented to produce enzymes. The specific operation process was as follows:

[0053] Inoculate the spores of Penicillium purpurea Li-3, T. pinophilum Li-93, Aspergillus terreus Li-20 and Aspergillus pylorus Li-62 in potato dextrose liquid culture medium respectively, at 30°C, 200rpm Incubate on a shaker for 48 hours. Then inoculated in the secondary seed liquid culture medium with 5% inoculum amount, cultivated on a shaker at 30° C. and 200 rpm for 24 hours to obtain the secondary seed liquids of the above-mentioned fungi.

[0054] Secondary seed liquid medium preparation method: 200g of peeled potatoes, add 1000mL of water, boil for about 30 minutes, filter through gauze, add 8g of glucose, dissolve and a...

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Abstract

The invention provide a fermentation medium and method for improving the activity of beta-glucuronidase produced by fungi. The fermentation medium is a solution prepared by filtering boiled potatoes and water and adding a promoter and water, wherein the promoter is glycyrrhizic acid or a salt thereof. According to the fermentation medium and method provided by the invention, Penicillium purpurogenum Li-3, Talaromyces pinophilum Li-93, Aspergillus terreus Li-20 and Aspergillus ustus Li-62 are subjected to fermentation and culture for production of beta-glucuronidase; and it is found that the fermentation enzyme activity unit of each fungus is increased by 100 to 600% and fermentation time is brought forward by 36 to 60 h compared with the prior art. Thus, the fermentation medium and method provided by the invention have the characteristics of simple operation, low cost, high fermentation enzyme activity unit and short enzyme production time.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a fermentation medium and a method for improving the enzyme activity of fungi producing β-glucuronidase. Background technique [0002] β-glucuronidase (β-glucuronidase, EC: 3.2.1.31) is a class of glycosidases that can catalyze the hydrolysis of β-glucuronidic acid bonds, and widely exists in human body, animals, plants and microorganisms. β-glucuronidase was first discovered to exist in various tissues and body fluids of humans and animals, especially in the liver, spleen and adrenal glands. Early people found that this enzyme was related to tumor invasion and metastasis. Now It has become an important means of tumor diagnosis and treatment by detecting the level of this enzyme in a specific part. In addition, based on the hydrolysis properties of this enzyme, some studies have shown that this enzyme can be used in tumor enzymatic prodrug-directed therapy. Many...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12R1/80C12R1/645C12R1/66C12R1/70
CPCC12N9/2402C12Y302/01031
Inventor 李春徐赢华贾锦彤吕波冯旭东
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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