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Method for separating and purifying catalpin in fermented liquid of catalpa endophytic fungi by extraction and back-extraction

A technology for purifying catalpin and endophytic fungi, applied in the field of biochemistry, can solve the problem of low purity of catalpin, and achieve the effects of preventing environmental pollution, high separation and purification ability, and safe operation process

Active Publication Date: 2017-01-04
成都上交致远生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the existence of a large number of residual medium components and impurities such as fermentation products in the fermentation broth, the purity of the biologically active component catalinin in the fermentation broth is relatively low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0032] The preparation of the endophytic fungus fermented liquid of Chinese catalpa comprises the following steps in the following examples:

[0033] 1. Isolation of endophytic fungi: splitting endophytic fungi and purifying them by mycelial tip cutting;

[0034] 2. Induce spore production: Inoculate Trichoderma longibrachiata CCTCC M 208102 on the PDA medium, cut off a piece of the PDA medium with mycelia, put it on the CMX medium, and cultivate it at room temperature. After 1 to 3 days, there will be spores grow;

[0035] 3. Liquid fermentation culture: pick the spores and transfer them to the PDA liquid medium, culture at 26-28° C. on a shaking table at 180 rpm, and stop the fermentation after 1-8 days.

Embodiment 1

[0037] This embodiment relates to a method for concentrating and separating catalpaline from the fermented liquid of endophytic fungus of catalpa, comprising the following steps:

[0038] (1) adjust the pH of 1 liter of fermentation broth to be 4;

[0039] (2) 1 liter of pH is 4 dilute hydrochloric acid aqueous solution to balance 1 liter of ethyl acetate for 30 minutes, after removing the aqueous solution, adding 1 liter of new pH again is 4 dilute hydrochloric acid aqueous solution to carry out saturated equilibrium to ethyl acetate, carry out 3 times repeatedly Sub-acidic water pre-saturation treatment, the test found that the pH of the water phase after equilibrium was 4;

[0040] (3) Mix the fermented broth with pH=4 in the previous two steps and ethyl acetate, shake and mix for 10 minutes, and then stand for half an hour to form a clear phase interface.

[0041] (4) After removing the raffinate aqueous phase, add 1 liter of new aqueous phase with pH=11, oscillate and mi...

Embodiment 2

[0046] This embodiment relates to a method for concentrating and separating catalpaline from the fermented liquid of endophytic fungus of catalpa, comprising the following steps:

[0047] (1) adjust the pH of 1 liter of fermented liquid to be 4.5;

[0048] (2) 0.5 liters of dilute hydrochloric acid aqueous solution that pH is 4.5 is balanced 30 minutes to 0.5 liters of ethyl acetate with 0.5 liters of pH, after removing the aqueous solution, add again 0.5 liters of new pHs and be that the dilute hydrochloric acid aqueous solution of 4.5 carry out saturated equilibrium to ethyl acetate, carry out 3 times repeatedly Sub-acidic water pre-saturation treatment, the test found that the pH of the water phase after equilibrium was 4.5;

[0049](3) Mix the fermented broth with pH = 4.5 in the previous two steps and ethyl acetate, shake and mix for 10 minutes, then let it stand for half an hour to form a clear phase interface;

[0050] (4) After removing the raffinate aqueous phase, ad...

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PUM

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Abstract

The invention discloses a method for separating and purifying catalpa bungei element in catalpa bungei endophytic fungi fermentation liquor by using extraction and back-extraction. The method comprises the following steps: adjusting the pH of the catalpa bungei endophytic fungi fermentation liquor to be 4-6; carrying out pre-saturation treatment on ethyl acetate by using dilute acid solution so as to enable the pH of the ethyl acetate to be matched with that of the fermentation liquor; uniformly mixing the catalpa bungei endophytic fungi fermentation liquor with the adjusted pH with the ethyl acetate, standing for double liquid phase extraction, and collecting ethyl acetate phase; uniformly mixing the collected ethyl acetate phase with water phase with the pH of 7-11, carrying out back-extraction, and recycling the water phase to obtain a purified catalpa bungei element aqueous solution. Compared with the prior art, by utilizing weak acidity of the catalpa bungei element, under the condition of guaranteeing that the yield of the catalpa bungei element is more than 80%, and through a simple extraction and back-extraction process, hydrophilic impurities and lyophobic impurities in the fermentation liquor are separated and removed, the purity of the catalpa bungei element is improved by more than ten times, the ethyl acetate can be recycled after suspended evaporation during the separation process, the separation cost is low, and the environmental pressure is low.

Description

technical field [0001] The present invention relates to the field of biochemistry, and relates to a method for separating and purifying catalpin from fermented liquid of endophytic fungus of catalpa and its application, in particular to a method for separating and purifying catalin in fermented liquid of endophytic fungus of catalpa by extraction and back extraction prime method. Background technique [0002] Endophytic fungi are those fungi that live inside plant tissues and do not cause obvious plant diseases. Endophytic fungi can produce the same or similar bioactive substances as the host plant, which has great potential value and can be a source of new anticancer drugs. Our research group has isolated and extracted an endophytic fungus (CCTCC M 208102) with strong anti-cancer activity from Juglans chinensis, and initially identified the fungus as Deuteromycotina, Hyphomycetes, Polychospora, Wood Mold belongs to the genus, and its fermented extract has a strong inhibit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07G99/00
Inventor 苗志奇冯婷婷胡郁楠于湘莉
Owner 成都上交致远生物科技有限公司