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Chlamydia spectabilis zrv2011f2‑p3 and its application

A technology of ZRV2011F2-P3, Clamydia spectabilis, applied in the direction of fungi, microorganisms, metabolic diseases, etc., to achieve the effect of hindering the occurrence and development, preventing and treating hypercholesterolemia

Active Publication Date: 2017-12-29
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, natural compounds with sEH inhibitory effects have not been reported yet.

Method used

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  • Chlamydia spectabilis zrv2011f2‑p3 and its application
  • Chlamydia spectabilis zrv2011f2‑p3 and its application
  • Chlamydia spectabilis zrv2011f2‑p3 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Isolation and identification of strain Chlamydia spectiosa ZRV2011F2-P3 and detection of sEH activity inhibitors in its metabolites

[0025] 1. Isolation and purification of strains

[0026]Filamentous fungi were isolated from rice vinegar solid-state fermented vinegar mash (cake from rice saccharification stage) using potato dextrose agar (PDA) medium. Use sterile water to prepare vinegar mash into a suspension with a mass concentration of 20%, further dilute it with sterile water 10 times, 100 times and 1000 times, spread it on tiger red PDA plates respectively, and incubate at 25°C-30°C for 5 days , From the plate that can pick out a single colony, select colonies of different forms and streak culture on the PDA plate, culture at 25°C-30°C for 3-5 days, pick a single colony from the plate for streak culture, and save it on the PDA slant . Pick colonies from the PDA slant and inoculate them on the PDA plate for streak culture, culture at 25-30°C for 3 day...

Embodiment 2

[0052] Embodiment 2 Chlamydia spectiosa ZRV2011F2-P3 inhibitor

[0053] (1) Inoculate Phytophthora spectiosa ZRV2011F2-P3 to PDA slant medium, cultivate at 25~30°C for 5 days, and obtain the thalline slant; the composition of the PDA medium: potato extract powder 10.0g / L, glucose 20.0g / L, agar 13g / L, chloramphenicol 0.1g / L, the solvent is deionized water, and the pH is natural.

[0054] (2) Inoculate the slant bacteria from step (1) into the M6 ​​medium, culture at 25-30°C for 11-13 days, take the culture medium and freeze-dry it (Freeze Dry System, LABCONCO, USA) for 24 hours, then add 1 times the volume of the culture medium 90% methanol aqueous solution (volume concentration), vortexed, placed in an ultrasonic cleaner (KUDOS, Shanghai Kedao) for 10 sec, leached at 4°C for 12 h, centrifuged the extract to obtain the supernatant, That is, the methanol extract is obtained, the methanol extract is concentrated under reduced pressure to remove the solvent, and the concentrate ...

Embodiment 3

[0055] Example 3. The inhibitory effect of Chlamydia spectabilis ZRV2011F2-P3 on sEH is enhanced with the increase of added concentration

[0056] 1. sEH enzyme activity detection conditions

[0057] (1) Detection of phosphatase activity

[0058] Control: with buffer (25mM bis-tris HCl, 1mM MgCl 2 , 0.1mg / ml BSA, pH7.0) as the reaction medium, human sEH enzyme with a final concentration of 300ng / ml was used as a catalyst, and the substrate Attophos (promega) with a final concentration of 5μM was added, and the reaction was carried out at 30°C for 30 minutes. Fluorescence dynamic detection was performed with an excitation wavelength of 435nm and an emission wavelength of 555nm.

[0059] experiment:

[0060] Use buffer (25mM bis-tris HCl, 1mM MgCl2, 0.1mg / ml BSA, pH 7.0) as the reaction medium, human sEH enzyme at a final concentration of 300ng / ml as a catalyst, and the substrate Attophos at a final concentration of 5μM (promega), and then add the sEH inhibitor obtained in E...

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Abstract

The invention discloses a new bacterial strain-Chlamydomonas spectabilis ZRV2011F2-P3 and its application as a soluble cyclooxygenase (sEH) activity inhibitor. Microbiology Center, the preservation number is CGMCC NO.10637, the preservation date is March 19, 2015, and the preservation address is No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, postcode 100101; this strain comes from traditional Fermented vinegar mash with rice vinegar can produce inhibitors of cardiovascular disease target sEH enzyme, strongly inhibit the activity of sEH N-terminal phosphatase, so as to prevent and treat hypercholesterolemia, and help to reveal cardiovascular disease target sEH new physiological functions.

Description

(1) Technical field [0001] The present invention relates to a kind of soluble cyclooxygenase inhibitor, relate in particular to a strain new bacterial strain---spectacular silk clothes mold (Talaromyces (Byssochlamys) spectabilis) ZRV2011F2-P3 (the sexual type of Paecilomyces variotii (Paecilomyces variotii)) and Its use as a soluble cyclooxygenase inhibitor. (2) Background technology [0002] Soluble cyclooxygenase (soluble epoxy hydrolase, sEH) is a new target of cardiovascular disease with cross-functionality that has emerged in recent years. The inhibitor of this enzyme is considered to be the most promising target drug for cardiovascular disease. Blood pressure, anti-inflammatory function, as well as the protection of myocardium, kidney, brain and other end organs related to hypertension and atherosclerosis (Marino, J.P., Jr. (2009) Soluble epoxy hydrolase, a target with multiple opportunities for cardiovascular drug discovery. Curr Top Med Chem, 9, 452-63.). [0003]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A61K36/06A61P3/06C12R1/645
Inventor 朱立颖王欣
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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