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Method for separating and purifying 7-hydroxyl tropolone from bacteria supernatant

A technology for the separation and purification of hydroxytropenolone, which is applied in the fields of bioengineering and biochemistry, can solve the problems of flammability, harmful reagents, and low recovery rate, and achieve simple and easy implementation of instrument reagents, avoiding cumbersome processes and toxicity The effect of reagent participation

Active Publication Date: 2017-04-19
WUHAN UNIV
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Problems solved by technology

[0004] 7-Hydroxytropenolone has great medicinal potential. At present, the main source is chemical synthesis, but there are problems such as flammability, explosion, and harmful reagents in chemical synthesis methods.
The existing microbial fermentation extraction of 7-hydroxytropenol also has problems such as low yield, low purity, and low recovery rate

Method used

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  • Method for separating and purifying 7-hydroxyl tropolone from bacteria supernatant
  • Method for separating and purifying 7-hydroxyl tropolone from bacteria supernatant

Examples

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Embodiment 1

[0025] Example 1 Isolation, purification and identification of 7-hydroxytropenolone

[0026] (1) Extraction and purification of 7-hydroxytropenolone

[0027] The medium for cultivating Pseudomonas donghu HT and purifying 7-hydroxytropenolone is MKB liquid medium. Each liter of MKB liquid medium contains ingredients: 15 mL glycerol, 3.28 g dipotassium hydrogen phosphate trihydrate, 5 g acid hydrolyzed casein, 2.5 g magnesium sulfate heptahydrate, pH 7-7.2; The total content of iron ions is not higher than 1 μM. Pseudomonas HT from East Lake was preserved in the China Center for Type Culture Collection (date of preservation: August 6, 2015, preservation address: China. Wuhan. Wuhan University), and its classification is named Pseudomonas donghuensis HT, the deposit number is CCTCC NO: M 2015481.

[0028] 1) Inoculate Pseudomonas HT in a test tube containing 5 mL of liquid MKB medium, and incubate overnight on a shaker at 30°C for activation.

[0029] 2) Transfer the overni...

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Abstract

The invention discloses a method for separating and purifying 7-hydroxyl tropolone from bacteria supernatant. The method comprises the following steps: (1) collecting fermentation liquor of pseudomonas donghuensis HT (the accession number is CCTCC NO: M 2015481), centrifuging and then filtering supernatant to obtain supernatant of a bacteria solution; (2) extracting the supernatant of the bacteria solution with ethyl acetate, retaining a water phase of the lower layer, regulating the pH of the water phase to be acidic, extracting the water phase with the ethyl acetate again, and collecting an organic phase of the upper layer; (3) evaporating the ethyl acetate in the organic phase to dryness under reduced pressure, and dissolving the rest of substances in anhydrous ethanol; and (4) carrying out chromatographic purification by using Sephadex LH-20, wherein a mobile phase is the anhydrous ethanol; collecting components with ferrous chelate activity, and evaporating the anhydrous ethanol to dryness under reduced pressure to obtain the 7-hydroxyl tropolone. By the method, the 7-hydroxyl tropolone of which the purity reaches 90% or above can be obtained, and the recovery rate is greater than 60%.

Description

technical field [0001] The invention belongs to the technical fields of bioengineering and biochemistry, and in particular relates to a method for separating and purifying 7-hydroxytropenolone from bacterial supernatant. Background technique [0002] 7-Hydroxytropenolone compounds are a class of natural products belonging to the tropotrienone-type compounds, which exhibit multiple biological activities in many aspects. Due to the addition of a hydroxyl group at the ortho position of the ketone group, 7-hydroxytropolone exhibits stronger inhibitory activity relative to tropolone. Some studies have shown that 7-hydroxytropenolone and its derivatives β-thujaplicinol (β-thujaplicinol), 3,7-dihydroxytropenolone, puberulic acid , Puberulonic acid can be used as a potential therapeutic drug for many diseases. 7-Hydroxytropenolone showed strong inhibitory activity against Gram-positive bacteria, such as Staphylococcus aureus ( Staphylococcus aureus ) and Micrococcus luteus ( Mi...

Claims

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Application Information

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IPC IPC(8): C07C49/717C07C45/78
CPCC07C45/78C07C49/717
Inventor 谢志雄姜桢
Owner WUHAN UNIV
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