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Phomopsis sp. strain and application thereof in performing biotransformation on celastrol

A technology of tripterine and Phomopsis, applied in the biological field, can solve the problems of affecting application, strong killing effect on normal cells, and low water solubility

Active Publication Date: 2019-07-02
NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although CSL has significant pharmacological activity, its low water solubility, high myocardial toxicity and strong killing effect on normal cells have seriously affected its clinical application.

Method used

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  • Phomopsis sp. strain and application thereof in performing biotransformation on celastrol
  • Phomopsis sp. strain and application thereof in performing biotransformation on celastrol
  • Phomopsis sp. strain and application thereof in performing biotransformation on celastrol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, acquisition, identification and preservation of bacterial strains

[0043] 1. Acquisition of strains

[0044] 1. Get the disease-free fresh leaves of Tripterygium wilfordii plant in Yao County, Dali, Yunnan Province, and carry out surface disinfection. Surface disinfection method: rinse with clean water, rinse 4 times with clean water, rinse with 75% alcohol for 30s, then soak in 5% sodium hypochlorite for 2s, rinse with sterile water 10 times.

[0045] 2. Sterile filter paper absorbs the moisture on the surface, cuts off the liquid-contact side with a sterilized blade, then cuts the inside into small pieces and transplants it into the modified Martin solid medium containing 150mg / L penicillin and 120mg / L streptomycin , and then pick the growing single colonies for culture.

[0046] 3. Further carry out continuous purification to obtain a purely cultured strain, which is named LGT-5 strain.

[0047] 2. Identification of strains

[0048] 1. Colony morph...

Embodiment 2

[0063] Embodiment 2, application Phomopsis LGT-5 prepares 16-OH-Celastrol

[0064] 1. Application of Phomopsis LGT-5 for tripterine transformation

[0065] 1. Inoculate Phomopsis LGT-5 into a Erlenmeyer flask containing 150mL of modified Martin liquid medium, and cultivate it with shaking at 28°C and 180rpm until the logarithmic growth phase (usually the culture time is 1 day), at this time Phomopsis The concentration of mold LGT-5 was 5 g dry weight / L culture system. Dry weight measurement method: sample the culture system, collect the bacteria by centrifugation, wash thoroughly with sterile water, and then dry to dry weight.

[0066] 2. After completing step 1, add 2 mL of tripterine solution (containing 50 mg of tripterine, the solvent is DMSO) to the culture system, and culture with shaking at 28° C. and 180 rpm for 5 days. That is, the ratio of Phomopsis LGT-5 and tripterine is: 0.75 g dry weight of Phomopsis LGT-5: 50 mg tripterine.

[0067] 3. After completing step 2...

Embodiment 3、1

[0077] Example 3, Functional Verification of 16-OH-Celastrol

[0078] Test compound: Celastrol or 16-OH-Celastrol prepared in Example 2.

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PUM

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Abstract

The invention discloses a phomopsis sp. strain and an application thereof in performing biotransformation on celastrol. The phomopsis sp. strain provided by the invention is phomopsis sp. LGT-5, and the preservation registration number is CGMCC No.16088. The application of the phomopsis sp. LGT-5 in a compound of a preparation formula (III) is protected. The phomopsis sp. LGT-5 can transform the celastrol into a new compound 16-OH-Celastrol. Compared with the celastrol, the 16-OH-Celastrol has no significant difference in antibacterial activity, but the cytotoxicity is greatly reduced, and the16-OH-Celastrol can be used as a new bacteriostatic drug.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a Phomopsis strain and its application in biotransformation of tripterine. Background technique [0002] Tripterygium wilfordii has antibacterial, anti-inflammatory, and anti-tumor effects. The active ingredient in Tripterygium wilfordii is Celastrol (CSL). The structural formula of tripterine is as follows: [0003] [0004] Although CSL has significant pharmacological activity, its low water solubility, high myocardial toxicity and strong killing effect on normal cells have seriously affected its clinical application. What needs to be solved at present is to improve or retain pharmacological activity while reducing toxicity so as to improve the therapeutic index. [0005] Biotransformation of natural active lead compounds is a method of using biological systems (such as: enzymes, microorganisms, plant cells) to modify the specific molecular structure of na...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P33/02C07J3/00A01N45/02A01P1/00A61K31/56A61P31/04C12R1/645
CPCA01N45/02A61K31/56A61P31/04C07J3/005C12P33/02C12N1/145C12R2001/645
Inventor 刘成吴秀丽钟翙依张彩芳刘河涛杨延辉王洒
Owner NINGXIA MEDICAL UNIV
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