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A kind of penicillium and method for producing fumagillin

A technology of fumagillin and penicillium, which is applied in the field of microbial engineering and can solve the problems of low fermentation level and the like

Active Publication Date: 2020-07-14
ZHEJIANG HISUN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This shows that the fumagillin-producing bacteria used in the prior art is relatively primitive, and the fermentation level is low, so it is necessary to find a new high-efficiency fumagillin-producing bacteria

Method used

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  • A kind of penicillium and method for producing fumagillin
  • A kind of penicillium and method for producing fumagillin
  • A kind of penicillium and method for producing fumagillin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: strain source

[0036] Penicillium HS-NF-684Z (CGMCC No.14144) is a Penicillium isolated from the soil of a hillside in Mount Qingcheng, Chengdu City, Sichuan Province, China.

[0037] Cross-sampled the soil in the Qingcheng Mountain area, randomly selected 5 sampling points, took 10g of soil samples from each point, put them into a conical flask, mixed them evenly, took 10g of samples, and added them to a conical flask filled with 90mL sterile water medium (there is a magnetic stirrer in the bottle), vortex and stir for 30 minutes to make it fully mixed to make a suspension, which is 10 -1 bacterial suspension. Dilute the above suspension with sterile water at a volume ratio of 1:9 to 10 by dilution coating plate method. -2 , 10 -3 , 10 -4 , 10 -5 Concentration, take 0.1mL of bacterial suspension with different dilution multiples, spread it on a Czapek–Dox Medium plate, gently spread it on the surface of the medium with a sterile applicator stick, an...

Embodiment 2

[0040] Embodiment 2: strain identification

[0041]Conduct experiments with reference to relevant contents in books such as "Mycology of China", "General Mycology", "Handbook of Common Bacterial System Identification", "Guidelines for Molecular Cloning Experiments" and "Chinese Pharmacopoeia" (2015 Edition); color judgment refers to RAL K7 The color of the color card series is compared.

[0042] 1. Strain number: HS-NF-684Z (CGMCC No.14144).

[0043] 2. The cultural characteristics of the bacterial strain: adopt CA medium (Czapek Agar) to cultivate at 25°C for 12 days, adopt CYA medium (Czapek Yeast Autolysate Agar) to cultivate at 5°C / 25°C / 37°C for 7 days, adopt G25N medium (25% Glycerol Gitrate Agar) was cultured at 25°C for 7 days, and the morphology, color and pigment of the mycelia were observed. The culture characteristics of the strains are shown in Table 1. Four kinds of media, PDA, SDA, MEA and MEPG, were used to culture at 25°C for 5 to 7 days, and the color and pi...

Embodiment 3

[0070] Example 3: 18s rDNA sequence analysis and strain identification

[0071] The experiments were carried out referring to the relevant content in the book "Molecular Cloning Experiment Guide". Collect the fresh bacteria of the strain, then use the fungal DNA extraction kit to extract the total DNA of the bacteria, and use the universal primer NS1 (SEQ ID NO: 1) / NS8 (SEQ ID NO: 2) to amplify the 18S rDNA sequence, the amplification system and The PCR reaction program is shown in Table 9. PCR products were detected by 0.8% agarose gel electrophoresis, and PCR products were purified and recovered by SanPrep column PCR purification product kit. The purified PCR products were directly sent to Nanjing KingScript Biotechnology Co., Ltd. Perform sequence determination.

[0072] Table 9 PCR amplification system and reaction program

[0073]

[0074] The measured 18s rDNA sequence (SEQ ID NO:3) was proofread and compared with the related species and genus sequences in the GenBa...

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Abstract

The invention discloses a new Penicillium sp. HS‑NF‑684Z, the preservation number is: CGMCC No.14144; meanwhile, it discloses the preparation of fumagillin or a drug combination containing fumagillin by culturing it way of things.

Description

technical field [0001] The invention relates to the technical field of microbial engineering, in particular to a new penicillium and its application in the preparation of fumagillin. Background technique [0002] Fumagillin (formula I) was initially used to kill microsporidiosis parasites in bees, and was later found to be useful in the treatment of microsporidiosis and / or cryptosporidiosis caused by intestinal infections in humans, and this The disease can be quite fatal in immunocompromised patients. In recent years, with the in-depth study of fumagillin and its analogs, it has been reported that fumagillin and its analogs have anti-angiogenic activity, which can be achieved by binding to human methionine aminopeptidase-2 ( MetAP-2) binding, thereby blocking the formation of blood vessels. Therefore, fumagillin and its analogs are widely used as angiogenesis inhibitors for the treatment of cancer. [0003] [0004] In the prior art, F.R.Hanson et al. reported that fu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P17/16A61K31/336A61P9/00A61P35/00C12R1/80
CPCA61K31/336A61P9/00A61P35/00C12N1/14C12P17/162C12N1/145C12R2001/80
Inventor 张延青钱娉婷熊兵邓爱文姜南李建宋张辉应灵萍王继栋滕云郑玲辉
Owner ZHEJIANG HISUN PHARMA CO LTD
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