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Method for separation and preparation of eburicoic acid compound by using hirschioporus versatilis fermentation liquor

A technology of fermented liquid and perforate, which is applied in the field of separation and preparation of perforate compounds, can solve the problems such as unreported chemical components, and achieve the effect of simple and easy medium preparation, large-scale production and short fermentation time

Inactive Publication Date: 2010-05-12
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, no report has been found on the fruiting body and the chemical composition of the fermented liquid of P.

Method used

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  • Method for separation and preparation of eburicoic acid compound by using hirschioporus versatilis fermentation liquor

Examples

Experimental program
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example 1

[0011] Using the method of liquid culture in a flat plate and shaker flask, first use 20g of glucose, 200g of peeled potatoes, MgSO 4 1.5g, KH 2 PO 4 3g, VB 1 10mg, add water, mix and stir until 1000ml to make a culture medium; then culture and ferment in the dark under the conditions of temperature: 25°C and rotation speed: 170r / min for 15 days to obtain a total of 10 liters of fermentation broth, which is divided into two parts: fermentation broth and mycelium The fermented liquid is extracted twice with ethyl acetate to obtain 2 gram extracts; the wet weight of mycelia is 420 grams, and the weight of 80 grams after drying; times, a total of 6 grams of the extract was obtained; after TLC detection, the two parts of the extract were combined, separated on a silica gel column, and eluted with a gradient of chloroform-methanol system with a volume ratio of 100-80:0-20, and each 100ml was a stream Parts; in the chloroform-methanol (volume ratio 95:5) elution part obtains a gr...

example 2

[0013] Using the method of liquid culture in a flat plate and shake flask, first use 15g of glucose, 180g of peeled potatoes, MgSO 4 1.0g, KH 2 PO 4 2.5g, VB 1 8 mg, mixed with water and stirred to 1000ml to make culture medium; then cultured and fermented in the dark under the conditions of temperature: 20°C and rotation speed: 150r / min for 12 days, and a total of about 10 liters of fermentation broth was obtained, which was divided into fermentation broth and mycelia. part; the fermentation broth was extracted twice with ethyl acetate to obtain 1.8 grams of extract; the wet weight of mycelium was 400 grams, and the weight of 75 grams after drying; the dried mycelia was extracted with chloroform-methanol (volume ratio 70:30) Twice, a total of 5.5 grams of extract was obtained; after TLC detection, the two parts of extract were combined, separated on a silica gel column, and eluted with a gradient of chloroform-methanol system with a volume ratio of 100-80:0-20, one for each...

example 3

[0015] Using the method of liquid culture in a flat plate and shake flask, first use 25g of glucose, 220g of peeled potatoes, MgSO 4 2.0g, KH 2 PO 4 3.2g, VB 1 12 mg, mixed with water and stirred to 1000ml to make culture medium; then cultured and fermented in the dark under the conditions of temperature: 32°C and speed: 200r / min for 18 days, and a total of 12 liters of fermentation broth was obtained, which was divided into two parts: fermentation broth and mycelia The fermented liquid is extracted twice with ethyl acetate to obtain 2.5 grams of extract; 450 grams of mycelia wet weight, 82 grams after drying; the dry mycelium is extracted twice with chloroform-methanol (volume ratio 65: 35) times, a total of 6.5 grams of extract was obtained; after TLC detection, two parts of extract were combined, separated on a silica gel column, and eluted with a gradient of chloroform-methanol system with a volume ratio of 100-80:0-20, and every 100ml was a stream Parts; in the chlorof...

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Abstract

The invention provides a method of separating and preparing eburicoic acid compound by the fermentation liquid of long hair follicle fungus. A plate transgenic shaking flask liquid culture method is adopted. Firstly, glucose, peeled potatoes, MgSO4, KH2PO4, VB1 and water are mixed and stirred to prepare culture medium; then, culturing the culture medium to be fermented to obtain fermentation liquor; extracting the fermentation liquid to obtain an extract; drying mycelium and extracting with chloroform-methanol to obtain extract; combining the two extracts for the column chromatography separation and the chloroform-methanol gradient elution; a group of mixtures containing the compound are obtained at the chloroform-methanol elution part after the repeated column chromatography; petroleum ether-acetone elution is adopted to separate the mixtures to obtain a compound eburicoic acid pure product. The invention can solves the raw material source problem for developing the eburicoic acid into raw medicine of drugs for curing human leukemia. And the invention has the advantages of mild culture conditions, easy medium preparation, short fermentation tie, large yield, easy production in large scale, etc.

Description

technical field [0001] The invention relates to a method for separating and preparing a dentoic acid compound by utilizing the fermented liquid of the folliculopore bacteria. Background technique [0002] According to the literature L.Francisco, Q.Jose, R.Augusto, et al., Lanostanoid triterpenes from Laetiporus sulphureus and apoptosis induction on HL-60 human myeloid leukemia cells.J.Nat.Prod., 2004, 67(12), 2008. , the compound peroracic acid can induce the apoptosis of human HL-60 myeloid leukemia cells, and its IC 50 =23-27μM, 3β-hydroxyacetylated dental acid showed the strongest effect of inducing apoptosis of human HL-60 myeloid leukemia cells (IC 50 = 13-15 μM). Its chemical structure is as follows: [0003] [0004] In view of the excellent activity of the compound and its acetylated compound in inducing the apoptosis of human HL-60 myeloid leukemia cells, it is worthwhile to further study the work of developing peroracic acid as a raw material for the treatmen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00A61P35/02C07J9/00C12R1/645
Inventor 麻兵继刘吉开
Owner HENAN AGRICULTURAL UNIVERSITY
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