A kind of preparation method and use of secondary metabolites of endophytic fungus chaetomium sp of Eucalyptus margina

A technology of secondary metabolites and endophytic fungi, applied in the direction of microorganism-based methods, biochemical equipment and methods, botany equipment and methods, etc.

Active Publication Date: 2022-04-19
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a preparation method and application of secondary metabolites of Eucalyptus endophyte Chaetomium sp. Problem, to isolate the secondary metabolites of Chaetomium sp., and the secondary metabolites have antibacterial effect

Method used

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  • A kind of preparation method and use of secondary metabolites of endophytic fungus chaetomium sp of Eucalyptus margina
  • A kind of preparation method and use of secondary metabolites of endophytic fungus chaetomium sp of Eucalyptus margina
  • A kind of preparation method and use of secondary metabolites of endophytic fungus chaetomium sp of Eucalyptus margina

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0050] Experimental example 1 Fermentation of endophytic fungus Chaetomium sp.

[0051] First, pick a little endophytic fungus Chaetomium sp. from the 4°C cryotube Inoculate on the PDA solid medium plate for activation and culture for 5-7 days, subculture for 4-5 days, then inoculate the mycelia in a 500mL Erlenmeyer flask filled with 200mL PDB liquid medium, place at 25°C, 150rpm Shake culture for 6-7 days. The obtained liquid seeds were inserted into the sterilized rice culture medium, and then transferred to a constant temperature incubator at 28°C for fermentation for 60 days, and a total of 6Kg of rice culture medium was fermented.

experiment example 2

[0052] Preparation of Experimental Example 2 Secondary Metabolites

[0053] (1) Preparation of ethyl acetate layer crude extract

[0054] The endophytic fungus Chaetomium sp. solid fermentation product was taken out from the fermentation bottle and placed in the extraction barrel, the fungus block was smashed up with a shovel, poured into an appropriate amount of methanol for cold extraction, and the extraction was repeated 3 times, each time for 3 days. The filtrate was obtained by filtering with gauze, the fermentation broth was vacuum filtered, and concentrated to dryness under reduced pressure to obtain a crude extract. The crude extract was suspended in water, extracted with petroleum ether and ethyl acetate, and then concentrated under reduced pressure to obtain the crude extract of petroleum ether layer (15.15 g) and the crude extract of ethyl acetate layer (51.40 g), respectively.

[0055] (2) column chromatography under reduced pressure of ethyl acetate layer crude e...

experiment example 3

[0077] The antibacterial activity of experimental example 3 secondary metabolites

[0078] (1) Test bacteria

[0079] The selected test bacteria were: Staphylococcus aureus (S.aureus ATCC29213 and drug-resistant strain S.aureus N50) (G + ), Escherichia coli (G - ), Agrobacterium tumefaciens ATCC 11158 (G - ), Pseudomonas lachrymansATCC 11921 (G - ), Ralstonia solanacearum (G - ) and tomato scab (Xanthomonasvesicatoria ATCC 11633) (G - ).

[0080] (2) Solution preparation

[0081] Phosphate buffer solution (PBS) configuration: use NaH 2 PO 4 and Na 2 HPO 4 Prepare 0.2 mol / L PBS buffer solution with pH=7.2.

[0082] Preparation of thiazolium blue (MTT) solution: the above-mentioned PBS buffer solution was prepared into a MTT solution with a concentration of 5 mg / mL.

[0083] Configuration of sample solution: Accurately weigh 2.0 mg of each secondary metabolite separated above and dissolve in 0.3 mL of acetone, then add 0.7 mL of distilled water to make a mother solut...

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Abstract

The invention discloses a preparation method and application of a secondary metabolite of the endophytic fungus Chaetomium sp., the method comprising: (1) activating and subculturing the endophytic fungus Chaetomium sp. on PDA solid medium, Then shake culture in PDB liquid medium; (2) ferment and cultivate the liquid seeds obtained in step (1) in rice medium to obtain solid fermented product; (3) solid fermented product is extracted by methanol cold immersion, filtered, The filtrate is concentrated to obtain a crude extract, and the crude extract is suspended in water and extracted with petroleum ether and ethyl acetate to obtain the crude extract of petroleum ether and ethyl acetate layer respectively; (4) by vacuum column chromatography, dextran Compound A‑F was obtained by gel column chromatography and semi-preparative high performance liquid chromatography. The method of the invention separates the secondary metabolites of the endophytic fungus Chaetomium sp., and the secondary metabolites have antibacterial and antitumor effects.

Description

technical field [0001] The invention relates to a secondary metabolite of the Eucalyptus endophyte fungus, in particular to a preparation method and application of a secondary metabolite of the endophytic fungus Chaetomium sp. Background technique [0002] Eucalyptus exserta F.Muell, also known as Eucalyptus exserta F.Muell, is one of the eucalyptus species introduced into my country from Australia earlier, and is widely planted in Guangdong, Hainan and Guangxi. In addition to being an important economic timber forest, the leaves of Eucalyptus aeruginosa can be used as Chinese herbal medicine in the folk, which has the effect of reducing blood pressure and anti-inflammation, and the methanol extract of its bark has insecticidal effect. [0003] With the large-scale planting of eucalyptus, the scope and degree of disease occurrence also increased. The occurrence of gray rot, bacterial wilt, scorch, canker and root rot has caused huge losses to the cultivation of eucalyptus. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/08C12P7/22A61K31/365A61K31/235A61P35/00A61P31/04A61P39/06A01N37/40A01P1/00C12R1/645
CPCA61P31/04A61P35/00A61P39/06A01N37/40A61K31/235A61K31/365C12P7/22C12P17/08
Inventor 单体江毛子翎欧阳锦逵王军孙思王伟
Owner SOUTH CHINA AGRI UNIV
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