Method of separating and purifying destruxins of Metarrhizium anisopliae

A technology for the separation and purification of metarhizin, which is applied in the field of separation and purification, can solve the problems of high preparation cost, achieve the effects of reducing preparation cost, reducing the amount of solvent, and saving metarhizin

Active Publication Date: 2007-08-15
GUANGDONG XINJINGXIANG BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, although a variety of metarhizines have been identified, most of them focus on structural analysis and determination of physical and chemical properties. As for the separation and preparat...

Method used

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  • Method of separating and purifying destruxins of Metarrhizium anisopliae
  • Method of separating and purifying destruxins of Metarrhizium anisopliae
  • Method of separating and purifying destruxins of Metarrhizium anisopliae

Examples

Experimental program
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Embodiment 1

[0024]After fermenting the liquid produced from Metarhizium anisopliae var.anisopliae, centrifuge it with a centrifuge at 5000-8000 rpm for 10-15 minutes to remove the slag; or vacuum filter to remove the slag; 10% hydrochloric acid solution to adjust the pH value of the fermentation broth to 4.0, use kaolin or diatomaceous earth as an adsorbent, and add it to the fermentation broth according to the weight of the adsorbent / volume of the fermentation broth = 2%, at 24 ° C, 150 rpm Oscillating the adsorption place for 1 hour; then centrifuging with a centrifuge at 5000 to 8000 rpm for 10 to 15 minutes to remove the adsorbent and retain the fermentation broth, as shown in Figure 2; use 10% hydrochloric acid solution to adjust the pH value of the fermentation broth to 4.0, the extraction solvent is a mixture of the following volume ratios: ethyl acetate: dichloromethane = 1: 1; the volume ratio of the extraction solvent to the fermentation broth is 50: 50; the ultrasonic wave with ...

Embodiment 2

[0026] Repeat the same steps as described in Example 1, but the conditions of adsorption method pretreatment of fermentation broth, ultrasonic wave to promote extraction, and preparative chromatographic separation and purification process are respectively changed to:

[0027] (1) Conditions for pretreatment of fermentation broth by adsorption method:

[0028] Use 10% hydrochloric acid solution to adjust the pH value of the fermentation broth to 3.0-6.0; the adsorbent is kaolin, diatomaceous earth, bentonite, or a mixture of one or both of them; the amount of adsorbent: adsorbent weight / fermentation broth volume = 1-5%; temperature: 15-35°C; oscillating speed: 100-200 rpm; adsorption treatment for 0.5-2 hours.

[0029] (2) Conditions for ultrasonic-promoted extraction:

[0030] Use 10% hydrochloric acid solution to adjust the pH value of the fermentation broth to 3.0-6.0; the extraction solvent is a mixed solution of the following volume ratio: ethyl acetate: dichloromethane=4...

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Abstract

The invention discloses a separating and purifying method of Metarrhizium anisopliae, which comprises the following steps: (1) adsorbing fungal ferment liquid to remove partial impurity; removing adsorbent through centrifuging; obtaining supernatant layer of ferment liquid; (2) extracting supernatant layer under ultrasound; separating organic phase and water phase through separatory funnel; reserving organic phase to condense into rough extract of Metarrhizium anisopliae; (3) decompressing rough extract of Metarrhizium anisopliae; condensing and drying fraction; dissolving and filtering; (4) making spectrum; separating and purifying the rough product in the high-effective liquid phased chromatograph to produce high-purity product.

Description

technical field [0001] The invention relates to a method for separation and purification, in particular to a method for separation and purification of metarhizine. technical background [0002] Metarhizium (destruxin) is a mycotoxin, is a class of cyclic decarboxypeptide compounds, mainly produced by Metarhizium, other fungi such as Aschersonia, Altemaria, single terminal Trichothecium and Ophiosphaerella can also form different types of metarhizine. Metarhizine has biological activities such as insecticidal, antiviral and anticancer, and has high economic value. Since Kodaira isolated metarhizine A and B for the first time in 1961, 35 metarhizines have been analyzed and identified. From Kodaira (1961), Susuki (1970) to Pail et al. (1981) all adopt the traditional classical method to isolate metarhizine. The first step of the method is extraction, the fermentation broth of Metarhizium anisopliae is centrifuged or filtered to remove slag, then its pH value is adjusted to a...

Claims

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Application Information

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IPC IPC(8): C07K1/36C07K1/16C07K14/37A61K36/06
Inventor 胡琼波任顺祥
Owner GUANGDONG XINJINGXIANG BIOLOGICAL ENG CO LTD
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