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Spectacular byssochlamys ZRV2011F2-P3 and application thereof

A technology of ZRV2011F2-P3 and Clamydia spectabilis, applied in the direction of fungi, microorganisms, metabolic diseases, etc., to prevent and treat hypercholesterolemia, hinder the occurrence and development of the effect

Active Publication Date: 2015-10-07
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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  • Spectacular byssochlamys ZRV2011F2-P3 and application thereof
  • Spectacular byssochlamys ZRV2011F2-P3 and application thereof
  • Spectacular byssochlamys ZRV2011F2-P3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Isolation and identification of strain S. spectabilis 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 of rice saccharification stage) using potato dextrose agar (PDA) medium. The vinegar mash was prepared into a suspension with a mass concentration of 20% by using sterile water, further diluted 10 times, 100 times and 1000 times with sterile water, respectively coated on the tiger red PDA plate, and cultivated at 25 ℃-30 ℃ for 5 days , From the plate that can pick out a single colony, select colonies of different shapes on the PDA plate for streak culture, cultivate at 25℃-30℃ for 3-5 days, select a single colony from the streak culture plate, and store it on the PDA slant. . The colonies were picked from the PDA slant and inoculated onto the PDA plate for streak culture, cultured at 25...

Embodiment 2

[0052] Example 2 S. spectacleum ZRV2011F2-P3 inhibitor

[0053] (1) Inoculate S. spectatorioides ZRV2011F2-P3 on PDA slant medium, cultivate at 25~30°C for 5 days to obtain bacterial slant; the composition of described 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 slanted cells in step (1) into 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 volume of the culture medium 90% methanol aqueous solution (volume concentration), mixed by vortex, placed in an ultrasonic cleaner (KUDOS, Shanghai Science and Technology Guide) for 10 sec, leached at 4 °C for 12 h, centrifuged the leaching solution to get the supernatant, That is, a methanol extract is obtained, and the methanol extract is concentrated under reduced pressure to remove the solvent to obtain a...

Embodiment 3

[0055] Example 3. The inhibitory effect of S. spectacleum ZRV2011F2-P3 on sEH was enhanced with the increase of the 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, the human sEH enzyme with a final concentration of 300ng / ml was used as a catalyst, 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 excitation wavelength of 435 nm and emission wavelength of 555 nm.

[0059] experiment:

[0060] Buffer (25mM bis-tris HCl, 1mM MgCl2, 0.1mg / ml BSA, pH 7.0) was used as the reaction medium, the final concentration of 300ng / ml of human sEH enzyme was used as the catalyst, and the final concentration of 5μM substrate Attophos was added (promega), and then add the sEH inhib...

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Abstract

The invention discloses a new bacterial strain-spectacular byssochlamys ZRV2011F2-P3 and application to serving as an active inhibitor of soluble epoxidase (sEH). The spectacular byssochlamys ZRV2011F2-P3 is preserved in China General Microbiological Culture Collection Center, the preserving serial number is CGMCC NO.10637, the preserving date is March 19, 2015, the preserving address is Institute of Microbiology of Chinese Academy of Sciences, number 3 of the first courtyard of west beichen road of the chaoyang district of Beijing, and the postcode is 100101; and the bacterial strain comes from traditional rice vinegar fermented vinegar mash, the inhibitor of cardiovascular disease target sEH enzyme can be produced, the activity of the N end phosphatase of the sEH is powerfully restrained, the effect of preventing and treating hypercholesteremia is achieved, and the new physiological function of the cardiovascular disease target sEH is easily disclosed.

Description

(1) Technical field [0001] The invention relates to a soluble cyclooxygenase inhibitor, in particular to a new strain--Talaromyces (Byssochlamys) spectabilis ZRV2011F2-P3 (sexual type of Paecilomyces variotii) and its use as a soluble cyclooxygenase inhibitor. (2) Background technology [0002] Soluble epoxide hydrolase (sEH) is a new target for cardiovascular disease with cross-functionality that has appeared in recent years. The inhibitor of this enzyme is considered to be the most promising target drug for cardiovascular disease, not only has anti-high Blood pressure, anti-inflammatory functions, and protection of end-organs such as myocardium, kidney, and brain related to hypertension and atherosclerosis (Marino, J.P., Jr. (2009) Soluble epoxide hydrolase, a target with multiple opportunities for cardiovascular drug discovery. Curr Top Med Chem, 9, 452-63.). [0003] sEH is a homodimer composed of two 60Kda monomers in an anti-parallel fashion with dual enzymatic activ...

Claims

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Application Information

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Patent Type & Authority Applications(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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