Preparation method for separating and extracting new compound from A. terreus secondary metabolite

A technology for secondary metabolites and compounds, applied in the field of separation and extraction of new compounds, can solve the problem that secondary metabolites are not new secondary metabolites, etc., and achieve the effects of improving extraction rate and accelerating the release of intracellular substances

Inactive Publication Date: 2019-01-29
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the fermentation process of the marine Penicillium and its secondary metabolite Flufuran has not u

Method used

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  • Preparation method for separating and extracting new compound from A. terreus secondary metabolite
  • Preparation method for separating and extracting new compound from A. terreus secondary metabolite
  • Preparation method for separating and extracting new compound from A. terreus secondary metabolite

Examples

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[0020] Example 1:

[0021] A preparation method for separating and extracting new compounds from A. terreus secondary metabolites, including spore preparation, strain fermentation, secondary metabolite extraction, and separation and purification. The specific steps are:

[0022] Spore preparation: Place the glycerol cryotube of the strain A.terreus OUCMDZ-2739 (containing 20% ​​glycerol and 3.3% seawater in a freshly prepared PDA liquid cryopreservation solution) stored in an ultra-low temperature refrigerator at -80°C in a clean operating table. Resuscitate for 10 minutes. Dip the resuscitated cryopreservation solution with the inoculating loop after burning and sterilization, and streak evenly in the freshly prepared PDA slope solid medium, and then place the slope medium in a 28℃ constant temperature incubator 4 Day, cultivate the strain slant to mature, get plump spores, set aside;

[0023] Strain fermentation: Add the epigenetic modification regulator TSA to the fungal culture ...

Example Embodiment

[0031] Example 2:

[0032] A preparation method for isolating and extracting new compounds from A. terreus secondary metabolites, including spore preparation, strain fermentation, secondary metabolite extraction, and separation and purification. The specific steps are:

[0033] Spore preparation: Place the glycerol cryopreservation tube of strain A.terreus OUCMDZ-2739 (containing 20% ​​glycerol and 3.3% seawater in a freshly prepared PDA liquid cryopreservation solution) stored in an ultra-low temperature refrigerator at -80°C in a clean operating table. Resuscitate for 10 minutes. Dip the resuscitated cryopreservation liquid with the inoculating loop after burning and sterilization, and streak evenly in the freshly configured PDA slope solid medium, then place the slope medium in a 28℃ constant temperature incubator for 3 Day, cultivate the strain slant to mature, get plump spores, set aside;

[0034] Strain fermentation: Add the epigenetic modification regulator TSA to the fungal ...

Example Embodiment

[0039] Example 3:

[0040] A preparation method for isolating and extracting new compounds from A. terreus secondary metabolites, including spore preparation, strain fermentation, secondary metabolite extraction, and separation and purification. The specific steps are:

[0041] Spore preparation: Place the glycerol cryopreservation tube of strain A.terreus OUCMDZ-2739 (containing 20% ​​glycerol and 3.3% seawater in a freshly prepared PDA liquid cryopreservation solution) stored in an ultra-low temperature refrigerator at -80°C in a clean operating table. Resuscitate for 10 minutes. Dip the resuscitated cryopreservation liquid with the inoculating loop after the burning and sterilization, and streak evenly in the freshly prepared PDA slope solid medium, and then place the slope medium in a 28℃ constant temperature incubator for culture 5 Day, cultivate the strain slant to mature, get plump spores, set aside;

[0042] Strain fermentation: Add the epigenetic modification regulator TSA ...

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Abstract

The invention discloses a preparation method for separating and extracting a new compound from an A. terreus secondary metabolite. The method comprises the steps of spore preparation, strain fermentation, extraction on the secondary metabolite, and separation and purification, specifically, an epigenetic modifying regulator TSA, nicotinamide and dipeptide are added into a fungal culture medium, fermentation culture is carried out, a fermentation broth and mycelia are subjected to separate extraction, and a fermentation concentrate is separated and purified by column chromatography and high performance liquid chromatography to obtain the separated and extracted new compound. The beneficial effects are as follows: the nicotinamide and dipeptide are added into the culture medium and produce asynergistic gain effect with the epigenetic modifying regulator TSA, and epigenetic modification is performed on A. terreus OUCMDZ-2739 to effectively activate the expression of the silencing gene ofthe strain, and obtain a new secondary metabolite through separation; the novel compound prepared by the invention has a good anti-tumor effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method for separating and extracting new compounds from secondary metabolites of A. terreus. Background technique [0002] As we all know, microorganisms have long been an important source of active substances such as antibiotics and enzyme inhibitors. Microorganisms have become a hotspot in the field of natural product research due to their large-scale fermentation and cultivation, no raw material restrictions, and no environmental damage. In recent years, the probability of obtaining new compounds from terrestrial microorganisms has been greatly reduced. Due to the unique living environment of high pressure, high salinity, low temperature, and oligotrophic conditions, marine-derived microorganisms have completely different metabolic pathways from terrestrial microorganisms. In addition to similar metabolites on land, it is easier to produce some structurally specific...

Claims

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Application Information

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IPC IPC(8): C07D311/04A61P35/00C12P17/06C12R1/66
CPCA61P35/00C12P17/06C07D311/04
Inventor 黄芳芳孙坤来朱伟明张译文王斌陈荫赵玉勤
Owner ZHEJIANG OCEAN UNIV
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