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Ginkgo endophytic fungus and application thereof

A technology of endophytic fungi and ginkgo, applied in the field of microorganisms, can solve the problems of slow growth of ginkgo, little anti-tumor activity, and few endophytic fungi of ginkgo

Active Publication Date: 2018-06-01
德州汇洋生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, ginkgo grows slowly, and it takes more than 20 years from planting to fruiting under natural conditions, which restricts the development of its medicinal efficacy. According to the endosymbiotic theory of plant endophytic fungi, it is very likely that there are some endophytic fungi in ginkgo, which can Produce the same or similar chemical components as Ginkgo biloba, so Ginkgo endophytes have become a research hotspot, and people expect it to become a new source of ginkgo drugs or a new way of bioactive substances
However, there are relatively few studies on the endophytic fungi of Ginkgo biloba, especially for the anti-tumor activity of endophytic fungi of Ginkgo biloba.

Method used

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  • Ginkgo endophytic fungus and application thereof
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  • Ginkgo endophytic fungus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Screening and Identification of Ginkgo Endophytic Fungi

[0024] Separation material: Ginkgo is collected from Linyi City, Shandong Province. The material part is the trunk bark about 2cm above the ground, with a thickness of about 3-8mm. The age of the ginkgo tree is about 30 years, and the trunk diameter is about 55cm. Take the collected fresh ginkgo bark, wash its surface with distilled water, remove the outer skin after slightly drying, cut the xylem into small sections of suitable length, soak in 75% ethanol for 6 minutes, and then rinse with sterile water for 2- 3 times, blot dry with sterile filter paper. Then cut it into small pieces and place it on the PDA medium containing streptomycin and penicillin, and culture it in a 25°C incubator for 3-20 days. After hyphae grow around the tissue, transfer it to a new PDA culture medium. On the basis, the culture was continued in a 25°C incubator. Until the grown colonies were identified as pure bacteria. T...

Embodiment 2

[0026] Example 2: Research on antitumor activity of endophytic fungi of Ginkgo biloba

[0027] Preparation of fermentation broth and mycelial crude extract: excavate three strains of Ginkgo biloba endophytic fungi cultured in PDA solid medium, inoculate the mycelium block into a conical flask with 250ml PDA liquid medium, 20°C, 120r / min shaker culture 7d. Then, ethyl acetate was added to each flask at a volume ratio of 1:1, and the culture was continued in a shaker for 4 days. After the fermentation liquid is filtered through 3-4 layers of gauze, the mycelium is removed, and the separation funnel is used for stratification to obtain the organic phase containing the secondary metabolites of the ginkgo endophytic fungus. A rotary evaporator is used to recover most of the ethyl acetate in the organic phase to obtain a concentrated solution containing secondary metabolites of endophytic fungi of Ginkgo biloba. Finally, use a vacuum concentration dryer to completely dry the conc...

Embodiment 3

[0031] Example 3 Research on Antitumor Active Substances of Ginkgo Endophytic Fungi

[0032] Semi-preparative HPLC method was used to analyze and separate the secondary metabolites of endophytic fungi of Ginkgo biloba: the chromatographic column was C18 column (250mmx10mm, 5μm), and the mobile phase was methanol-water. Elution with 20% methanol, volume flow rate 1.0ml / min, detection wavelength 210nm, column temperature 35°C, injection volume 100μl. According to the peak shapes of secondary metabolites of different strains, different substances were separated. Each substance was co-cultured with cervical cancer HeLa cells, and the effective anti-cervical cancer tumor active substances were determined by MTT method.

[0033] The secondary metabolites of strains J-1 and J-3 were separated and determined by semi-preparative HPLC, and the separation results were as follows: Figure 4 and Figure 5 As shown, from the semi-preparative HPLC peak profile ( Figure 4 and Figure 5 ...

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Abstract

The invention discloses a ginkgo endophytic fungus and an application thereof. The ginkgo endophytic fungus fusarium proliferatum DZHQ1 with anti-tumor activity is separated from ginkgo bark, the species of a strain is judged with a colony morphology and 18sRNA sequencing combination method, the anti-cervical cancer activity of a crude extract of the strain is detected with MTT assay, and finally,secondary metabolites of the strain with inhibition rate being higher than 50% are separated with a semi-preparative HPLC method. Accordingly, compound monomers with anti-tumor activity can be further screened.

Description

technical field [0001] The invention belongs to the field of microorganisms and tumor treatment, and in particular relates to a ginkgo endophytic fungus and its application in antitumor. Background technique [0002] Cancer is one of the major diseases that endanger human health today and seriously threatens human life. Cervical cancer is the most common gynecological malignancy, and its incidence tends to be younger in recent years. At present, there are more than 50 kinds of antineoplastic drugs commonly used clinically, but most of them can only alleviate the disease, and cannot achieve the purpose of complete cure. Therefore, the research and development of new anti-tumor drugs has always been the main aspect in the field of drug research. [0003] Ginkgo biloba is a special medicinal plant in my country. It has been found that Ginkgo biloba contains more than 100 chemical components, mainly including flavonoids and terpene lactones. Ginkgoxanthin and polyprenol in its...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P1/02A61K36/06A61P35/00C12R1/77
CPCA61K36/06C12N1/14C12P1/02C12N1/145C12R2001/77
Inventor 何庆曾强成王韬王海岛蒙建州朱清华焦德杰刘帅
Owner 德州汇洋生物科技有限公司
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