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Method for separation preparation of compound 2,4-dihydroxy-5-methyl-acetophenone by using Basidiomycetes

A technology of fermentation broth and porous bacteria, applied in botany equipment and methods, chemicals for biological control, fermentation, etc., to achieve the effects of reducing fermentation costs, improving product quality, and mild culture conditions

Inactive Publication Date: 2012-07-04
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Related literature [A.A.Morais, R.B.Fo, S.V.Fraiz Jr.Synthesis of threenatural 1,3-diarylpropanes: Two revised structures, Phytochemistry, 1989, 28(1): 239-242] and [Cai Mengshen, Wang Lanming. Carbon Glycoside Research-XXVI. Synthesis of Isomigoldin Analogs, Acta Chem. Sinica, 1990, 48, 1191-1198] only reported 2,4-dihydroxy-5-methyl- Acetophenone is used as a synthetic intermediate; but there is no report on its activity as a natural product and anti-phytopathogenic fungi
The applicant once reported for the first time the preliminary study on the antibacterial activity of the fermented metabolites of P. brown polypore (Dong Yanhong, Zhang Anling, Ma Huini, Li Xiaoming, Gao Jinming, Journal of Northwest Forestry University, 2007, 22 (2): 105-108), but, Antimicrobial active compounds in fermentation broth are unknown

Method used

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  • Method for separation preparation of compound 2,4-dihydroxy-5-methyl-acetophenone by using Basidiomycetes
  • Method for separation preparation of compound 2,4-dihydroxy-5-methyl-acetophenone by using Basidiomycetes
  • Method for separation preparation of compound 2,4-dihydroxy-5-methyl-acetophenone by using Basidiomycetes

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preparation Embodiment 1

[0033] This embodiment adopts the method for liquid culture of flat plate spinner bottle, and its concrete steps are as follows:

[0034] 1) Fermentation of bacterial strains and concentration of fermented liquid

[0035] Take 8-10 mm from the activated solid plate bacteria (28°C, cultured for 5-6 days). 2 2 to 3 pieces of mushroom cakes of different sizes were inserted into liquid PDA medium made of peeled potatoes 200g / L, glucose 20g / L, distilled water 1000mL, pH 7.2, and sterilized at 121°C for 30 minutes (the volume of liquid is 50mL / 150mL Erlenmeyer flask), at 28°C on a rotary shaker (rotational speed: 150r / min), cultured for 3 to 4 days to form bacterial balls, and then transferred to a fermentation bottle with a 10% inoculation amount (the liquid volume is 150mL / 500mL Erlenmeyer flask), fermented 13d, obtained fermented liquid 50L altogether. Concentrate 50L of fermented broth under reduced pressure at 55°C and halve it;

[0036] 2) Preparation of crude extract

[...

preparation Embodiment 2

[0046] The present embodiment adopts the method for the liquid culture of plate spinner bottle, and its specific steps are as follows:

[0047] 1) Fermentation of bacterial strains and concentration of fermented liquid

[0048] Take 6~8mm 2 Insert 3 to 4 pieces of mushroom cakes of different sizes into the liquid PDA medium made from 180g / L of peeled potatoes, 18g / L of glucose, 1000mL of distilled water, about pH7.2, and sterilized at 121°C for 30min. 150mL Erlenmeyer flask), cultivated in a rotary shaker (rotating speed: 130r / min) at 25°C for 4 to 5 days to form bacterial balls, and then transferred to a fermentation bottle with a 10% inoculum amount (a liquid volume of 130mL / 500mL) Erlenmeyer flask), fermented 14d, obtained fermented liquid 10L altogether. Concentrate 10L of fermentation broth at 55°C under reduced pressure and reduce it to half;

[0049] 2) Preparation of crude extract

[0050] The concentrated solution of the fermented product was extracted with ethyl ...

preparation Embodiment 3

[0054] The present embodiment adopts the method for the liquid culture of plate spinner bottle, and its specific steps are as follows:

[0055] 1) Fermentation of bacterial strains and concentration of fermented liquid

[0056] Take 6~8mm 2 Insert 2 to 3 pieces of mushroom cakes of different sizes into the liquid PDA medium made of peeled potatoes 250g / L, glucose 25g / L, distilled water 1000mL, pH7. / 150mL Erlenmeyer flask), cultivated in a rotary shaker (170r / min) at 34°C for 3 to 4 days to form bacterial balls, and then transferred to a fermentation flask with a 10% inoculation amount (the liquid volume is 170mL / 500mL Erlenmeyer flask) ), fermented 11d, and obtained a total of 15L of fermented liquid. Concentrate 15L of fermentation broth under reduced pressure at 55°C and reduce it to half;

[0057] 2) Preparation of crude extract

[0058] The concentrated solution of the fermented product was extracted with ethyl acetate, concentrated under reduced pressure to obtain 4....

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Abstract

The invention relates to a method for preparing a compound of 2, 4-dihydroxy-5-methyl-hypnone by separating the fermentation liquid of Polyporus picipes. In the method, a method of plate-to-flask liquid culture is adopted; firstly, peeled potato, glucose and distilled water with the pH of 7.2 approximately are sterilized and made into a culture medium; then the culture medium is cultured and fermented to obtain the fermentation liquid; the fermentation liquid is extracted by ethyl acetate to obtain extractum; the extractum is separated by column chromatography and is eluted by a chloroform-methanol system in a gradient way; and the pure compound of the 2, 4-dihydroxy-5-methyl-hypnone (II) is obtained after several times of silica gel column chromatography, reverse C18 column and SephadexLH-20 gel column chromatography and separation by a recrystallization method. The invention can develop the 2, 4-dihydroxy-5-methyl-hypnone (II) into the original drug of antipathogen fungi antibiotic and solves the problem of the raw material source of the 2, 4-dihydroxy-5-methyl-hypnone, and the invention has the advantages of mild culture condition, being simple and easy for confecting the culture medium, and short fermentation time, etc.

Description

technical field [0001] The invention relates to a preparation method of a compound, in particular to a method for separating and preparing compound 2,4-dihydroxy-5-methyl-acetophenone (II) by using the fermented liquid of Polyporus morphobacterium. The compound 2,4-dihydroxy-5-methyl-acetophenone (II) prepared by the present invention is a new natural product, which can inhibit the growth of various plant pathogenic fungi, and is effective against cotton fusarium wilt, Curvularia maize, Apple anthracnose bacteria have a good inhibitory effect. Background technique [0002] Higher fungi include some species of Ascomycetes and Basidiomycetes fungi. Due to long-term co-evolution, many higher fungi have formed their own unique chemical defense system. Once injured, the system will start to secrete enzymes to convert secondary metabolites into active substances. An interesting phenomenon is that the fruiting bodies of many macrofungi are never attacked by insects. A typical ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C49/825C12P7/26A01N35/04A01P3/00
Inventor 高锦明李勇解思达张鞍灵石新卫张炜玲
Owner NORTHWEST A & F UNIV
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