Preparation and application of chaetomium globosum and metabolite flavipin thereof

A technology of aflatoxin and Chaetomium globosa, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problem of unreported antioxidant activity and achieve strong antioxidant activity

Inactive Publication Date: 2013-05-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Chaetomium globosum CDW7 involved in this paper is a plant endophyte isolated from healthy Ginkgo biloba leaves, and its secondary metabolites, flavipin, 1,2-benzonedicarboxaldehyde-3,4,5-trihydroxy-6- methyl) was first isolated from Aspergillus flavipes and Aspergillus terreus by Raistrick. It has certain antibacterial and nematicide activity, but its antioxidant activity has not been reported so far.

Method used

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  • Preparation and application of chaetomium globosum and metabolite flavipin thereof
  • Preparation and application of chaetomium globosum and metabolite flavipin thereof
  • Preparation and application of chaetomium globosum and metabolite flavipin thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Example 1. Separation and purification of endophytic strain CDW7:

[0019] Collect fresh and healthy ginkgo leaves, wash them and soak them in 75% alcohol for 30 seconds, rinse them with sterile water 3 times, then soak them in 3-5% NaClO solution for 3-5 minutes, and rinse them with sterile water 3-5 times. Under aseptic conditions, cut the surface-sterilized ginkgo leaves into small pieces of 0.5×0.5cm, and place them in PDA medium and cultivate them at a constant temperature of 25°C. After the bacteria grow out, pick the edge hyphae of the colony for purification and culture Until the colony morphology is consistent, it is stored on the PDA slope to obtain the ginkgo endophyte. One of the strains is numbered CDW7 and is now deposited in the China Common Microorganism Collection Management Center (CGMCC for short). The preservation date is October 15, 2012, and the preservation number is CGMCC No. 6658.

Embodiment 2

[0020] Example 2. Identification of endophytic strain CDW7:

[0021] The endophytic fungus CDW7 is on the PDA flat medium, the colony is white at the beginning (see attached figure 1 ), gradual to dark cyan (see attached figure 2 ), the back is brown with irregular edges. Ascus shell scattered or grouped, ovoid or nearly spherical, size (418-610)μm×(222-277)μm (see attached image 3 ); The ascus shell wall is composed of dark brown interlaced filamentous tissue, the top hair has no septum, brown, tortuous or spirally curled, with fine spines on the surface, and the base diameter is 3.1μm; the ascospore is brown, subspherical or lemon-shaped, Size (7.5-10.2)μm×(6.5-7.5)μm, with a terminal bud hole (see attached Figure 4 ). The morphological characteristics of the above CDW7 are consistent with the morphological characteristics of Chaetomium sp. fungi.

[0022] Pick the plate-activated CDW7 mycelium block into the PDB medium, culture at 25°C for 5 days, and extract the total DNA u...

Embodiment 3

[0024] Example 3. Preparation of the compound flavipin:

[0025] (1) Fermentation culture of CDW7: Inoculate the CDW7 strain of Chaetomium globosum preserved at 4°C on a PDA plate, culture in a constant temperature incubator at 25°C for 5 days, and take 12 bacterial cakes with a diameter of 0.5 cm on the plate The blocks were put into a 250 mL Erlenmeyer flask containing 100 mL of PDB medium that had been sterilized, and cultured at 25° C. and 150 rpm for 14 days.

[0026] (2) Obtaining the crude extract of fermented product: take the fermentation broth obtained in step (1) above, separate the bacteria and fermentation broth with gauze to obtain 2L of total fermentation filtrate, and extract the fermentation filtrate with methylene chloride solvent for 3-5 Next, the extracts were combined and concentrated under reduced pressure to obtain crude fermented extract F1 (2g).

[0027] (3) Separation and purification of compounds: The crude extract of fermented product obtained in step (2)...

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Abstract

The invention provides preparation and application of chaetomium globosum and metabolite flavipin thereof. The strain is endophytic fungi obtained by separating gingko leaves, is identified to be chaetomium globosum with a number of CDW7, and is preserved in the China General Microbiological Culture Collection Center with the register number of CGMCC No. 6658. The invention relates to preparation and application of CDW7 metabolite flavipin at the same time. The preparation method comprises the step that the organic solvent extract of the strain CDW7 fermentation broth is subjected to gel column chromatography and recrystallization. The flavipin has strong anti-oxidation activity, and can be used as an antioxidant for application.

Description

Technical field [0001] The invention relates to the preparation and application of chaetomium globosum and its metabolite aflatoxin. Background technique [0002] More and more research results of free radical biology and free radical medicine indicate that the occurrence and development of cardiovascular diseases, tumors, senile dementia, aging, diabetes, AIDS and other diseases are closely related to reactive oxygen species and free radicals. The development of new, high-efficiency and safe natural antioxidants is one of the important research topics in the prevention and treatment of major diseases closely related to reactive oxygen species and free radicals. [0003] Plant endophytes refer to those fungi or bacteria that live in various tissues and organs of healthy plants at a certain stage or all stages of their life history. The infected host plant (at least temporarily) does not show external symptoms. The endogenous growth of microbial DNA can be proved by the method of h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/00A61K31/11A61P39/06C12R1/645
Inventor 叶永浩肖玉李红霞李枞刘俊彦
Owner NANJING AGRICULTURAL UNIVERSITY
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