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Extraction method of pyrrole-2-carboxylic acid

An extraction method and carboxylic acid technology, applied in the field of microorganisms, can solve the problems of limited extraction technology of pyrrole-2-carboxylic acid and the like

Inactive Publication Date: 2018-09-21
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, provide a kind of extraction method of pyrrole-2-carboxylic acid, aim to solve the technical problem that the extraction technology of existing pyrrole-2-carboxylic acid is very limited

Method used

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  • Extraction method of pyrrole-2-carboxylic acid

Examples

Experimental program
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Embodiment 1

[0029] Arenibacter sp.6A1 strain fermentation

[0030] The low-temperature preserved marine bacteria Arenibactersp.6A1 was revived on the plate medium, and then the revived strain was inoculated into the fermentation medium, 180rpm, 28°C for 96 hours;

[0031]Among them, the plate medium is 2216E liquid medium (recipe: peptone 5g / L, yeast extract 1g / L, ferric phosphate 0.01g / L, sea salt 20g / L) plus 1.5% agar, sterilized under high pressure at 121°C for 20 minutes The fermentation medium is 2216E liquid medium (formulation: peptone 5g / L, yeast extract 1g / L, ferric phosphate 0.01g / L, sea salt 20g / L).

[0032] After fermenting according to the above method, the fermentation broth of marine bacteria Arenibactersp.6A1 containing pyrrole-2-carboxylic acid can be obtained.

Embodiment 2

[0034] Isolation of pyrrole-2-carboxylic acid

[0035] The fermented liquid that embodiment 1 obtains is carried out following steps:

[0036] Ethyl acetate extraction: Use 1mol / L hydrochloric acid to adjust the pH of the fermented liquid to 3-4, extract with ethyl acetate, spin the extracted extract to dryness below 50°C, and repeat the extraction process three times to obtain acetic acid 2 g of the extract of the ethyl ester layer.

[0037] Gel column chromatography: dissolve the extract of the ethyl acetate layer in 2mL methanol, carry out Sephadex LH20 (column specification: diameter=3.5cm, column length=100cm) column chromatography, utilize 400mL volume ratio to be 1:1 The mixture of chloroform and methanol was used as the elution solvent for isocratic elution, each 50mL was collected as a fraction, and a total of 8 fractions were obtained, G3-A, G3-B, G3-C, G3-D, G3-E , G3-F, G3-G and G3-H.

[0038] HPLC separation: the G3-H fraction was concentrated and evaporated to...

Embodiment 3

[0046] Activity identification of pyrrole-2-carboxylic acid against plant pathogenic fungi

[0047] Antibacterial tests were carried out on 47 strains of clinical pathogenic bacteria by setting gradient concentrations of pyrrole-2-carboxylic acid. The pyrrole-2-carboxylic acid extracted in Example 2 was dissolved in DMSO to a sample concentration of 200 μg / μl, and then diluted to 100 μg / μl. 47 strains of clinical pathogenic bacteria were resuscitated (37°C, 1d) in LB (formulation: tryptone 10g / l, yeast extract 5g / l, NaCl 10g / l, agar 15g / l) plate medium, and then picked Take a single colony into a 50ml centrifuge tube containing 20ml of LB medium, culture it on a shaker at 180rpm at 37°C for 1 day; Evenly, pour the plate, place the sterilized filter paper on the solidified plate, add 2ul of the previously prepared pyrrole-2-carboxylic acid sample to the final concentration of the sample to 400μg / ml, 200μg / ml, add 2μl DMSO to the control group, Three repetitions were set up fo...

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Abstract

The invention belongs to the field of microbial technology, and specifically relates to an extraction method of pyrrole-2-carboxylic acid. The extraction method comprises the following steps: inoculating marine bacteria Arenibactersp.6Al into a fermentation culture medium for fermental cultivation to obtain a fermentation liquor; adjusting the fermentation liquor until the pH is equal to 3 to 4, and then carrying out extraction treatment to obtain an extractant; dissolving the extractant, carrying out gel column chromatography and HPLC separation in sequence to obtain pyrrole-2-carboxylic acid, wherein the conditions of HPLC separation are as follows: gradient elution is carried out by using a methyl alcohol / water mixed solvent, the detection wavelength is 254 nm, and the time for collecting a liquid-phase peak is 23.05 min. According to the method, the pyrrole-2-carboxylic acid with high purity and bacteriostatic activity is finally obtained, and a preferable basis is provided for industrial production of the pyrrole-2-carboxylic acid.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for extracting pyrrole-2-carboxylic acid. Background technique [0002] As the problem of drug resistance of harmful bacteria becomes more and more serious, the development of new and effective antibacterial active substances becomes more and more important. It is becoming more and more difficult to explore new active substances from terrestrial microorganisms. Relatively speaking, the ocean has become a more promising exploration target due to its environmental diversity and particularity. [0003] Bacteria of marine origin produce a variety of substances with antimicrobial activity. The more studied strains include Bacillus sp., Vibrio sp., Pseudomonassp., Flavobacteriumsp. and so on. Li Houjin et al. isolated prodigiosin from Pseudomonas bacteria from Daya Bay, which can effectively inhibit Gram-positive bacteria (Staphylococcus aureus and Bacillu...

Claims

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Application Information

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IPC IPC(8): C07D207/34C12P17/10A01N43/36A01P1/00A01P3/00C12R1/01
CPCA01N43/36C07D207/34C12P17/10
Inventor 王立岩伍嘉慧李晓帆林培玲
Owner SHENZHEN UNIV
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